In vitro test for the quantitative determination of haptoglobin in human serum and plasma on Roche/Hitachi cobas c systems.
Immunoturbidimetric assay.
Human haptoglobin forms a precipitate with a specific antiserum which is determined turbidimetrically.
Measuring range
0.1‑5.7 g/L (1.0‑57 µmol/L, 10‑570 mg/dL)
Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.
Lower limits of measurement
Lower detection limit of the test
0.1 g/L (1.0 µmol/L, 10 mg/dL)
The lower detection limit represents the lowest measurable analyte level that can be distinguished from zero. It is calculated as the value lying 3 standard deviations above that of the lowest standard (standard 1 + 3 SD, repeatability, n = 21).
Values below the lower detection limit (< 0.1 g/L) will not be flagged by the instrument.
0.3‑2.0 g/L (3.0‑20.0 µmol/L, 30‑200 mg/dL)
Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.
Criterion: Recovery within ± 10 % of the initial value at a haptoglobin concentration of 0.3 g/L (3.0 µmol/L, 30 mg/dL).
Icterus:
Hemolysis:
The Glick model which is normally used for assessment of hemoglobin interference is not suitable in the case of haptoglobin. Binding of free hemoglobin is the physiological function of haptoglobin. In the Glick study, hemolysate is added to the sample resulting in the formation of the haptoglobin‑hemoglobin complex. This complex is present in the reagent tube and causes a 10‑15 % decrease in haptoglobin values. However, the effect is of no relevance for the results in native samples because in vivo the haptoglobin‑hemoglobin complex is rapidly eliminated from the circulation and is practically not present in the blood.
Lipemia (Intralipid):
Rheumatoid factors up to 250 IU/mL do not interfere.
High dose hook‑effect: No false result occurs up to a haptoglobin concentration of 12 g/L (120 µmol/L, 1200 mg/dL).
Drugs: No interference was found at therapeutic concentrations using common drug panels.
In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.
For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.
ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on cobas c systems. All special wash programming necessary for avoiding carry‑over is available via the cobas link, manual input is required in certain cases. The latest version of the carry‑over evasion list can be found with the NaOHD/SMS/SmpCln1+2/SCCS Method Sheet and for further instructions refer to the operator’s manual.
Where required, special wash/carry‑over evasion programming must be implemented prior to reporting results with this test.
|
| Analyzer(s) on which cobas c pack(s) can be used | |
05992001 190 | Tina‑quant Haptoglobin ver.2 150 tests | System‑ID 03 9009 5 | cobas c 701/702 |
Materials required (but not provided):
11355279 216 | Calibrator f.a.s. Proteins (5 x 1 mL) | Code 656 | |
11355279 160 | Calibrator f.a.s. Proteins (5 x 1 mL, for USA) | Code 656 | |
10557897 122 | Precinorm Protein (3 x 1 mL) | Code 302 | |
10557897 160 | Precinorm Protein (3 x 1 mL, for USA) | Code 302 | |
11333127 122 | Precipath Protein (3 x 1 mL) | Code 303 | |
11333127 160 | Precipath Protein (3 x 1 mL, for USA) | Code 303 | |
05117003 190 | PreciControl ClinChem Multi 1 (20 x 5 mL) | Code 391 | |
05947626 190 | PreciControl ClinChem Multi 1 (4 x 5 mL) | Code 391 | |
05947626 160 | PreciControl ClinChem Multi 1 (4 x 5 mL, for USA) | Code 391 | |
05117216 190 | PreciControl ClinChem Multi 2 (20 x 5 mL) | Code 392 | |
05947774 190 | PreciControl ClinChem Multi 2 (4 x 5 mL) | Code 392 | |
05947774 160 | PreciControl ClinChem Multi 2 (4 x 5 mL, for USA) | Code 392 | |
05172152 190 | Diluent NaCl 9 % (119 mL) | System‑ID 08 6869 3 |
HAPT2: ACN 8228
Ready for use
cobas c 701/702 test definition | |||
Assay type | 2 Point End | ||
Reaction time / Assay points | 10 / 18‑38 | ||
Wavelength (sub/main) | 700/340 nm | ||
Reaction direction | Increase | ||
Units | g/L (µmol/L, mg/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 110 µL | – | |
R3 | 50 µL | – | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (NaCl) | ||
Normal | 5.5 µL | 9 µL | 180 µL |
Decreased | 5.5 µL | 4 µL | 164 µL |
Increased | 5.5 µL | 15 µL | 150 µL |
HAPT2 | |
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label. |
On‑board in use and refrigerated on the analyzer: | 12 weeks |
On‑board on the Reagent Manager: | 24 hours |
Diluent NaCl 9 % | |
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label. |
On‑board in use and refrigerated on the analyzer: | 4 weeks |
On‑board on the Reagent Manager: | 24 hours |
Calibrators | S1: H2O S2‑S6: C.f.a.s. Proteins | |
Multiply the lot-specific C.f.a.s. Proteins calibrator value by the factors below to determine the standard concentrations for the 6‑point calibration curve: | ||
S2: 0.0955 | S5: 1.45 | |
S3: 0.382 | S6: 2.28 | |
S4: 0.840 | ||
Calibration mode | RCM2 | |
Calibration frequency | Full calibration |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against the certified reference material in human serum of the IRMM (Institute for Reference Materials and Measurements) ERM-DA470k/IFCC.
Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.
Precision was determined using human samples and controls in an internal protocol with repeatability (n = 21) and intermediate precision (3 aliquots per run, 1 run per day, 21 days). The following results were obtained:
Repeatability | Mean g/L (µmol/L, mg/dL) | SD g/L (µmol/L, mg/dL) | CV % |
Precinorm Protein | 1.15 (11.5, 115) | 0.02 (0.2, 2) | 1.7 |
Precipath Protein | 1.98 (19.8, 198) | 0.03 (0.3, 3) | 1.5 |
Human serum A | 2.25 (22.5, 225) | 0.04 (0.4, 4) | 1.6 |
Human serum B | 5.04 (50.4, 504) | 0.06 (0.6, 6) | 1.1 |
Human serum C | 0.177 (1.77, 17.7) | 0.005 (0.05, 0.5) | 2.7 |
Intermediate precision | Mean g/L (µmol/L, mg/dL) | SD g/L (µmol/L, mg/dL) | CV % |
Precinorm Protein | 1.04 (10.4, 104) | 0.01 (0.1, 1) | 1.2 |
Precipath Protein | 1.73 (17.3, 173) | 0.02 (0.2, 2) | 1.1 |
Human serum 3 | 1.05 (10.5, 105) | 0.01 (0.1, 1) | 1.2 |
Human serum 4 | 1.57 (15.7, 157) | 0.02 (0.2, 2) | 1.2 |
Results for intermediate precision were obtained on the master system cobas c 501 analyzer.
Haptoglobin values for human serum and plasma samples obtained on a cobas c 701 analyzer (y) were compared with those determined using the corresponding reagent on a cobas c 501 analyzer (x).
Sample size (n) = 128
Passing/Bablok LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790. | Linear regression |
y = 0.964x + 0.026 g/L | y = 0.950x + 0.052 g/L |
τ = 0.983 | r = 0.999 |
The sample concentrations were between 0.110 and 5.43 g/L |
Haptoglobin is a transport and acute phase protein which is synthesized in hepatocytes. It is a glycoprotein which consists of two light α‑chains and two heavy β‑chains. The genetic polymorphism of the α‑chains leads to three phenotypes Hp 1‑1, Hp 2‑1 and Hp 2‑2 differing in molecular weight.
Haptoglobin binds hemoglobin in a strong haptoglobin‑hemoglobin complex (Hp‑Hb), the hemoglobin resulting from pathologically elevated hemolysis. These complexes are deposited in the hepatocytes, the deposition process having a half‑life of less than 10 minutes. Hemoglobin is enzymatically metabolized and haptoglobin is liberated after approximately 3 days. Complex formation and the extremely rapid elimination from circulating blood prevent the occurrence of hemoglobinuria with excess renal loss of iron. A reduction in the level of free haptoglobin is indicative of intravascular hemolysis.
As a strong positive acute phase reactant, a hemolysis‑mediated reduction or, to a certain extent, an elevation with accompanying acute inflammation can be compensated for. Indications for haptoglobin assays have been published and include the assessment of the severity and stage of intravascular hemolysis,
Various methods including nephelometry, radial immunodiffusion (RID) and turbidimetric methods are available for the determination of haptoglobin. The haptoglobin assay from Roche is based on the principle of immunological agglutination.
R1 | Phosphate buffer: 12.7 mmol/L, pH 7.2; NaCl: 130 mmol/L; PEG: 40 g/L; preservative |
R3 | Anti‑human haptoglobin antibody (rabbit): > 1.1 g/L; NaCl: 100 mmol/L; preservative |
R1 is in position B and R3 is in position C.
For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.
Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.
Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.
Safety data sheet available for professional user on request.
For USA: Caution: Federal law restricts this device to sale by or on the order of a physician.
For quality control, use control materials as listed in the \"Order information\" section.
In addition, other suitable control material can be used.
The control intervals and limits should be adapted to each laboratory’s individual requirements. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.
Follow the applicable government regulations and local guidelines for quality control.
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin and K2‑EDTA plasma
The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.
Centrifuge samples containing precipitates before performing the assay.
See the limitations and interferences section for details about possible sample interferences.
Stability: LREFTöpfer G, Hornig F, Sauer K, et al. Untersuchungen zur Stabilität von 11 Serumproteinen bei Bestimmung mittels Immunturbidimetrie. J Lab Med 2000;24(3):118-125. | 3 months at 15‑25 °C |
8 months at 2‑8 °C |
Sample stability claims were established by experimental data by the manufacturer or based on reference literature and only for the temperatures/time frames as stated in the method sheet. It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory.
In vitro test for the quantitative determination of haptoglobin in human serum and plasma on Roche/Hitachi cobas c systems.
Immunoturbidimetric assay
Human haptoglobin forms a precipitate with a specific antiserum which is determined turbidimetrically.
0.1‑5.7 g/L (1.0‑57 µmol/L, 10‑570 mg/dL)
Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.
Limit of Blank | = 0.1 g/L (1.0 µmol/L, 10 mg/dL) |
Limit of Detection | = 0.1 g/L (1.0 µmol/L, 10 mg/dL) |
Limit of Quantitation | = 0.1 g/L (1.0 µmol/L, 10 mg/dL) |
The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.
The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.
The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.
The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).
The Limit of Quantitation is the lowest analyte concentration that can be reproducibly measured with a total error of 20 %. It has been determined using low concentration haptoglobin samples.
0.3‑2.0 g/L (3.0‑20.0 µmol/L*, 30‑200 mg/dL*)
*calculated by unit conversion factor
Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.
Criterion: Recovery within ± 10 % of initial value at a haptoglobin concentration of 0.3 g/L.
Icterus:
Hemolysis:
The Glick model which is normally used for assessment of hemoglobin interference is not suitable in the case of haptoglobin. Binding of free hemoglobin is the physiological function of haptoglobin. In the Glick study, hemolysate is added to the sample resulting in the formation of the haptoglobin-hemoglobin complex. This complex is present in the reagent tube and causes a 10‑15 % decrease in haptoglobin values. However, the effect is of no relevance for the results in native samples because in vivo the haptoglobin-hemoglobin complex is rapidly eliminated from the circulation and is practically not present in the blood.
Lipemia (Intralipid):
Rheumatoid factors: No significant interference from rheumatoid factors up to a concentration of 250 IU/mL.
High-dose hook effect: No false result occurs up to a haptoglobin concentration of 12 g/L.
Drugs: No interference was found at therapeutic concentrations using common drug panels.
In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.
For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.
ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on cobas c systems. All special wash programming necessary for avoiding carry-over is available via the cobas link. The latest version of the carry-over evasion list can be found with the
|
| Analyzer(s) on which cobas c pack(s) can be used | |
08106045190 | Tina-quant Haptoglobin ver.2 (200 tests) | System‑ID 2064 001 | |
11355279160 | Calibrator f.a.s. Proteins (5 × 1 mL) | Code 20656 | |
10557897160 | Precinorm Protein (3 × 1 mL) | Code 20302 | |
11333127160 | Precipath Protein (3 × 1 mL) | Code 20303 | |
05947626160 | PreciControl ClinChem Multi 1 (4 × 5 mL) | Code 20391 | |
05947774160 | PreciControl ClinChem Multi 2 (4 × 5 mL) | Code 20392 | |
08063494190 | Diluent NaCl 9 % (123 mL) | System‑ID 2906 001 |
HAPT2: ACN 20640
Ready for use
Reporting time | 10 min | ||
Wavelength (sub/main) | 700/340 nm | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 78 µL | – | |
R3 | 36 µL | – | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (NaCl) | ||
Normal | 3.9 µL | 5 µL | 100 µL |
Decreased | 3.9 µL | 2 µL | 82 µL |
Increased | 3.9 µL | 5 µL | 100 µL |
For further information about the assay test definitions refer to the application parameters setting screen of the corresponding analyzer and assay.
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label. |
On-board in use and refrigerated on the analyzer: | 26 weeks |
Calibrators | S1: H2O S2-S6: C.f.a.s. Proteins |
Calibration mode | Non-linear |
Calibration frequency | Full calibration |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against the certified reference material in human serum of the IRMM (Institute for Reference Materials and Measurements) ERM‑DA470k/IFCC.
Representative performance data on the analyzers are given below. These data represent the performance of the analytical procedure itself.
Results obtained in individual laboratories may differ due to heterogenous sample materials, aging of analyzer components and mixture of reagents running on the analyzer.
Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP05‑A3 requirements with repeatability (n = 84) and intermediate precision (2 aliquots per run, 2 runs per day, 21 days).
Repeatability | Mean | SD | CV |
PCCC1a) | 0.780 (78.0) | 0.00469 (0.469) | 0.6 |
PCCC2b) | 1.38 (138) | 0.0114 (1.14) | 0.8 |
Human serum 1 | 0.250 (25.0) | 0.00354 (0.354) | 1.4 |
Human serum 2 | 1.13 (113) | 0.00880 (0.880) | 0.8 |
Human serum 3 | 1.90 (190) | 0.0178 (1.78) | 0.9 |
Human serum 4 | 2.81 (281) | 0.0268 (2.68) | 1.0 |
Human serum 5 | 4.99 (499) | 0.0487 (4.87) | 1.0 |
Intermediate precision | Mean | SD | CV |
PCCC1 FREFPreciControl ClinChem Multi 1 | 0.796 (79.6) | 0.0146 (1.46) | 1.8 |
PCCC2 FREFPreciControl ClinChem Multi 2 | 1.38 (138) | 0.0205 (2.05) | 1.5 |
Human serum 1 | 0.250 (25.0) | 0.0151 (1.51) | 6.1 |
Human serum 2 | 1.15 (115) | 0.0172 (1.72) | 1.5 |
Human serum 3 | 1.90 (190) | 0.0248 (2.48) | 1.3 |
Human serum 4 | 2.81 (281) | 0.0337 (3.37) | 1.2 |
Human serum 5 | 5.08 (508) | 0.0540 (5.40) | 1.1 |
Haptoglobin values for human serum and plasma samples obtained on a cobas c 503 analyzer (y) were compared with those determined using the corresponding reagent on a cobas c 501 analyzer (x).
Passing/Bablok LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790. | Linear regression |
y = 1.012x + 0.007 g/L | y = 0.971x + 0.0462 g/L |
τ = 0.989 | r = 0.999 |
The sample concentrations were between 0.110 and 5.62 g/L (11.0 and 562 mg/dL).
Haptoglobin values for human serum and plasma samples obtained on a cobas c 303 analyzer (y) were compared with those determined using the corresponding reagent on a cobas c 501 analyzer (x).
Passing/Bablok LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790. | Linear regression |
y = 1.014x + 0.0161 g/L | y = 0.969x + 0.0586 g/L |
τ = 0.977 | r = 0.999 |
The sample concentrations were between 0.150 and 5.60 g/L (15.0 and 560 mg/dL).
Haptoglobin is a transport and acute phase protein which is synthesized in hepatocytes. It is a glycoprotein which consists of two light α‑chains and two heavy β‑chains. The genetic polymorphism of the α‑chains leads to three phenotypes Hp 1‑1, Hp 2‑1 and Hp 2‑2 differing in molecular weight.
Haptoglobin binds hemoglobin in a strong haptoglobin‑hemoglobin complex (Hp‑Hb), the hemoglobin resulting from pathologically elevated hemolysis. These complexes are deposited in the hepatocytes, the deposition process having a half-life of less than 10 minutes. Hemoglobin is enzymatically metabolized and haptoglobin is liberated after approximately 3 days. Complex formation and the extremely rapid elimination from circulating blood prevent the occurrence of hemoglobinuria with excess renal loss of iron. A reduction in the level of free haptoglobin is indicative of intravascular hemolysis.
As a strong positive acute phase reactant, a hemolysis-mediated reduction or, to a certain extent, an elevation with accompanying acute inflammation can be compensated for. Indications for haptoglobin assays have been published and include the assessment of the severity and stage of intravascular hemolysis and evaluation of acute inflammatory processes.
Various methods including nephelometry, radial immunodiffusion (RID) and turbidimetric methods are available for the determination of haptoglobin. The haptoglobin assay from Roche is based on the principle of immunological agglutination.
R1 | Phosphate buffer: 12.7 mmol/L, pH 7.2; NaCl: 130 mmol/L; PEG: 40 g/L; preservative |
R3 | Anti‑human haptoglobin antibody (rabbit): > 1.1 g/L; NaCl: 100 mmol/L; preservative |
R1 is in position B and R3 is in position C.
For in vitro diagnostic use.
Exercise the normal precautions required for handling all laboratory reagents.
Disposal of all waste material should be in accordance with local guidelines.
Safety data sheet available for professional user on request.
For USA: Caution: Federal law restricts this device to sale by or on the order of a physician.
For quality control, use control materials as listed in the \"Order information\" section.
In addition, other suitable control material can be used.
The control intervals and limits should be adapted to each laboratory’s individual requirements. It is recommended to perform quality control always after lot calibration and subsequently at least every 26 weeks. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.
Follow the applicable government regulations and local guidelines for quality control.
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Serum
Plasma: Li‑heparin and K2‑EDTA plasma
The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.
Centrifuge samples containing precipitates before performing the assay.
See the limitations and interferences section for details about possible sample interferences.
Stability: LREFTöpfer G, Hornig F, Sauer K, et al. Untersuchungen zur Stabilität von 11 Serumproteinen bei Bestimmung mittels Immunturbidimetrie. J Lab Med 2000;24(3):118-125. | 3 months at 15‑25 °C 8 months at 2‑8 °C |
Sample stability claims were established by experimental data by the manufacturer or based on reference literature and only for the temperatures/time frames as stated in the method sheet. It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory.
In vitro test for the quantitative determination of haptoglobin in human serum and plasma on Roche/Hitachi cobas c systems.
Immunoturbidimetric assay
Human haptoglobin forms a precipitate with a specific antiserum which is determined turbidimetrically.
0.1‑5.7 g/L (1.0‑57 µmol/L)
Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.
Limit of Blank | = 0.1 g/L (1.0 µmol/L) |
Limit of Detection | = 0.1 g/L (1.0 µmol/L) |
Limit of Quantitation | = 0.1 g/L (1.0 µmol/L) |
The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.
The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.
The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.
The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).
The Limit of Quantitation is the lowest analyte concentration that can be reproducibly measured with a total error of 20 %. It has been determined using low concentration haptoglobin samples.
0.3‑2.0 g/L (3.0‑20.0 µmol/L*)
*calculated by unit conversion factor
Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.
Criterion: Recovery within ± 10 % of initial value at a haptoglobin concentration of 0.3 g/L.
Icterus:
Hemolysis:
The Glick model which is normally used for assessment of hemoglobin interference is not suitable in the case of haptoglobin. Binding of free hemoglobin is the physiological function of haptoglobin. In the Glick study, hemolysate is added to the sample resulting in the formation of the haptoglobin-hemoglobin complex. This complex is present in the reagent tube and causes a 10‑15 % decrease in haptoglobin values. However, the effect is of no relevance for the results in native samples because in vivo the haptoglobin-hemoglobin complex is rapidly eliminated from the circulation and is practically not present in the blood.
Lipemia (Intralipid):
Rheumatoid factors: No significant interference from rheumatoid factors up to a concentration of 250 IU/mL.
High-dose hook effect: No false result occurs up to a haptoglobin concentration of 12 g/L.
Drugs: No interference was found at therapeutic concentrations using common drug panels.
In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.
For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.
ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on cobas c systems. All special wash programming necessary for avoiding carry-over is available via the cobas link. The latest version of the carry-over evasion list can be found with the NaOHD/SMS/SCCS Method Sheet for information. For further instructions refer to the operator’s manual.
|
| Analyzer(s) on which cobas c pack(s) can be used | |
08106045 190 | Tina-quant Haptoglobin ver.2 (200 tests) | System‑ID 2064 001 | cobas c 503 |
Materials required (but not provided):
11355279 216 | Calibrator f.a.s. Proteins (5 × 1 mL) | Code 20656 | |
11355279 160 | Calibrator f.a.s. Proteins (5 × 1 mL, for USA) | Code 20656 | |
10557897 122 | Precinorm Protein (3 × 1 mL) | Code 20302 | |
10557897 160 | Precinorm Protein (3 × 1 mL, for USA) | Code 20302 | |
11333127 122 | Precipath Protein (3 × 1 mL) | Code 20303 | |
11333127 160 | Precipath Protein (3 × 1 mL, for USA) | Code 20303 | |
05117003 190 | PreciControl ClinChem Multi 1 (20 × 5 mL) | Code 20391 | |
05947626 190 | PreciControl ClinChem Multi 1 (4 × 5 mL) | Code 20391 | |
05947626 160 | PreciControl ClinChem Multi 1 (4 × 5 mL, for USA) | Code 20391 | |
05117216 190 | PreciControl ClinChem Multi 2 (20 × 5 mL) | Code 20392 | |
05947774 190 | PreciControl ClinChem Multi 2 (4 × 5 mL) | Code 20392 | |
05947774 160 | PreciControl ClinChem Multi 2 (4 × 5 mL, for USA) | Code 20392 | |
08063494 190 | Diluent NaCl 9 % (123 mL) | System‑ID 2906 001 |
HAPT2: ACN 20640
Ready for use
Reporting time | 10 min | ||
Wavelength (sub/main) | 700/340 nm | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 78 µL | – | |
R3 | 36 µL | – | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (NaCl) | ||
Normal | 3.9 µL | 5 µL | 100 µL |
Decreased | 3.9 µL | 2 µL | 82 µL |
Increased | 3.9 µL | 5 µL | 100 µL |
For further information about the assay test definitions refer to the application parameters setting screen of the corresponding analyzer and assay.
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label. |
On-board in use and refrigerated on the analyzer: | 26 weeks |
Calibrators | S1: H2O S2-S6: C.f.a.s. Proteins |
Calibration mode | Non-linear |
Calibration frequency | Full calibration |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against the certified reference material in human serum of the IRMM (Institute for Reference Materials and Measurements) ERM‑DA470k/IFCC.
Representative performance data on the analyzers are given below. These data represent the performance of the analytical procedure itself.
Results obtained in individual laboratories may differ due to heterogenous sample materials, aging of analyzer components and mixture of reagents running on the analyzer.
Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP05‑A3 requirements with repeatability (n = 84) and intermediate precision (2 aliquots per run, 2 runs per day, 21 days). The following results were obtained:
Repeatability | Mean | SD | CV |
PCCC1a) | 0.780 | 0.00469 | 0.6 |
PCCC2b) | 1.38 | 0.0114 | 0.8 |
Human serum 1 | 0.250 | 0.00354 | 1.4 |
Human serum 2 | 1.13 | 0.00880 | 0.8 |
Human serum 3 | 1.90 | 0.0178 | 0.9 |
Human serum 4 | 2.81 | 0.0268 | 1.0 |
Human serum 5 | 4.99 | 0.0487 | 1.0 |
Intermediate precision | Mean | SD | CV |
PCCC1 FREFPreciControl ClinChem Multi 1 | 0.796 | 0.0146 | 1.8 |
PCCC2 FREFPreciControl ClinChem Multi 2 | 1.38 | 0.0205 | 1.5 |
Human serum 1 | 0.250 | 0.0151 | 6.1 |
Human serum 2 | 1.15 | 0.0172 | 1.5 |
Human serum 3 | 1.90 | 0.0248 | 1.3 |
Human serum 4 | 2.81 | 0.0337 | 1.2 |
Human serum 5 | 5.08 | 0.0540 | 1.1 |
Haptoglobin values for human serum and plasma samples obtained on a cobas c 503 analyzer (y) were compared with those determined using the corresponding reagent on a cobas c 501 analyzer (x).
Passing/Bablok LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790. | Linear regression |
y = 1.012x + 0.007 g/L | y = 0.971x + 0.0462 g/L |
τ = 0.989 | r = 0.999 |
The sample concentrations were between 0.110 and 5.62 g/L.
Haptoglobin is a transport and acute phase protein which is synthesized in hepatocytes. It is a glycoprotein which consists of two light α‑chains and two heavy β‑chains. The genetic polymorphism of the α‑chains leads to three phenotypes Hp 1‑1, Hp 2‑1 and Hp 2‑2 differing in molecular weight.
Haptoglobin binds hemoglobin in a strong haptoglobin‑hemoglobin complex (Hp‑Hb), the hemoglobin resulting from pathologically elevated hemolysis. These complexes are deposited in the hepatocytes, the deposition process having a half-life of less than 10 minutes. Hemoglobin is enzymatically metabolized and haptoglobin is liberated after approximately 3 days. Complex formation and the extremely rapid elimination from circulating blood prevent the occurrence of hemoglobinuria with excess renal loss of iron. A reduction in the level of free haptoglobin is indicative of intravascular hemolysis.
As a strong positive acute phase reactant, a hemolysis-mediated reduction or, to a certain extent, an elevation with accompanying acute inflammation can be compensated for. Indications for haptoglobin assays have been published and include the assessment of the severity and stage of intravascular hemolysis,
Various methods including nephelometry, radial immunodiffusion (RID) and turbidimetric methods are available for the determination of haptoglobin. The haptoglobin assay from Roche is based on the principle of immunological agglutination.
R1 | Phosphate buffer: 12.7 mmol/L, pH 7.2; NaCl: 130 mmol/L; PEG: 40 g/L; preservative |
R3 | Anti‑human haptoglobin antibody (rabbit): > 1.1 g/L; NaCl: 100 mmol/L; preservative |
R1 is in position B and R3 is in position C.
For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.
Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.
Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.
Safety data sheet available for professional user on request.
For USA: Caution: Federal law restricts this device to sale by or on the order of a physician.
For quality control, use control materials as listed in the “Order information” section. In addition, other suitable control material can be used.
The control intervals and limits should be adapted to each laboratory’s individual requirements. It is recommended to perform quality control always after lot calibration and subsequently at least every 26 weeks. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.
Follow the applicable government regulations and local guidelines for quality control.
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Serum
Plasma: Li‑heparin and K2‑EDTA plasma
The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.
Centrifuge samples containing precipitates before performing the assay.
See the limitations and interferences section for details about possible sample interferences.
Stability: LREFTöpfer G, Hornig F, Sauer K, et al. Untersuchungen zur Stabilität von 11 Serumproteinen bei Bestimmung mittels Immunturbidimetrie. J Lab Med 2000;24(3):118-125. | 3 months at 15‑25 °C 8 months at 2‑8 °C |
Sample stability claims were established by experimental data by the manufacturer or based on reference literature and only for the temperatures/time frames as stated in the method sheet. It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory.
In vitro test for the quantitative determination of haptoglobin in human serum and plasma on Roche/Hitachi cobas c systems.
Immunoturbidimetric assay
Human haptoglobin forms a precipitate with a specific antiserum which is determined turbidimetrically.
0.1‑5.7 g/L (1.0‑57 µmol/L)
Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.
Limit of Blank | = 0.1 g/L (1.0 µmol/L) |
Limit of Detection | = 0.1 g/L (1.0 µmol/L) |
Limit of Quantitation | = 0.1 g/L (1.0 µmol/L) |
The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.
The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.
The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.
The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).
The Limit of Quantitation is the lowest analyte concentration that can be reproducibly measured with a total error of 20 %. It has been determined using low concentration haptoglobin samples.
0.3‑2.0 g/L (3.0‑20.0 µmol/L*)
*calculated by unit conversion factor
Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.
Criterion: Recovery within ± 10 % of initial value at a haptoglobin concentration of 0.3 g/L.
Icterus:
Hemolysis:
The Glick model which is normally used for assessment of hemoglobin interference is not suitable in the case of haptoglobin. Binding of free hemoglobin is the physiological function of haptoglobin. In the Glick study, hemolysate is added to the sample resulting in the formation of the haptoglobin-hemoglobin complex. This complex is present in the reagent tube and causes a 10‑15 % decrease in haptoglobin values. However, the effect is of no relevance for the results in native samples because in vivo the haptoglobin-hemoglobin complex is rapidly eliminated from the circulation and is practically not present in the blood.
Lipemia (Intralipid):
Rheumatoid factors: No significant interference from rheumatoid factors up to a concentration of 250 IU/mL.
High-dose hook effect: No false result occurs up to a haptoglobin concentration of 12 g/L.
Drugs: No interference was found at therapeutic concentrations using common drug panels.
In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.
For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.
ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on cobas c systems. All special wash programming necessary for avoiding carry-over is available via the cobas link. The latest version of the carry-over evasion list can be found with the NaOHD/SMS/SCCS Method Sheet for information. For further instructions refer to the operator’s manual.
|
| Analyzer(s) on which cobas c pack(s) can be used | |
08106045190 | Tina-quant Haptoglobin ver.2 (200 tests) | System‑ID 2064 001 |
Materials required (but not provided):
11355279216 | Calibrator f.a.s. Proteins (5 × 1 mL) | Code 20656 | |
10557897122 | Precinorm Protein (3 × 1 mL) | Code 20302 | |
11333127122 | Precipath Protein (3 × 1 mL) | Code 20303 | |
05117003190 | PreciControl ClinChem Multi 1 (20 × 5 mL) | Code 20391 | |
05947626190 | PreciControl ClinChem Multi 1 (4 × 5 mL) | Code 20391 | |
05117216190 | PreciControl ClinChem Multi 2 (20 × 5 mL) | Code 20392 | |
05947774190 | PreciControl ClinChem Multi 2 (4 × 5 mL) | Code 20392 | |
08063494190 | Diluent NaCl 9 % (123 mL) | System‑ID 2906 001 |
HAPT2: ACN 20640
Ready for use
Reporting time | 10 min | ||
Wavelength (sub/main) | 700/340 nm | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 78 µL | – | |
R3 | 36 µL | – | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (NaCl) | ||
Normal | 3.9 µL | 5 µL | 100 µL |
Decreased | 3.9 µL | 2 µL | 82 µL |
Increased | 3.9 µL | 5 µL | 100 µL |
For further information about the assay test definitions refer to the application parameters setting screen of the corresponding analyzer and assay.
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label. |
On-board in use and refrigerated on the analyzer: | 26 weeks |
Calibrators | S1: H2O S2-S6: C.f.a.s. Proteins |
Calibration mode | Non-linear |
Calibration frequency | Full calibration |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against the certified reference material in human serum of the IRMM (Institute for Reference Materials and Measurements) ERM‑DA470k/IFCC.
Representative performance data on the analyzers are given below. These data represent the performance of the analytical procedure itself.
Results obtained in individual laboratories may differ due to heterogenous sample materials, aging of analyzer components and mixture of reagents running on the analyzer.
Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP05‑A3 requirements with repeatability (n = 84) and intermediate precision (2 aliquots per run, 2 runs per day, 21 days).
Repeatability | Mean | SD | CV |
PCCC1a) | 0.780 | 0.00469 | 0.6 |
PCCC2b) | 1.38 | 0.0114 | 0.8 |
Human serum 1 | 0.250 | 0.00354 | 1.4 |
Human serum 2 | 1.13 | 0.00880 | 0.8 |
Human serum 3 | 1.90 | 0.0178 | 0.9 |
Human serum 4 | 2.81 | 0.0268 | 1.0 |
Human serum 5 | 4.99 | 0.0487 | 1.0 |
Intermediate precision | Mean | SD | CV |
PCCC1 FREFPreciControl ClinChem Multi 1 | 0.796 | 0.0146 | 1.8 |
PCCC2 FREFPreciControl ClinChem Multi 2 | 1.38 | 0.0205 | 1.5 |
Human serum 1 | 0.250 | 0.0151 | 6.1 |
Human serum 2 | 1.15 | 0.0172 | 1.5 |
Human serum 3 | 1.90 | 0.0248 | 1.3 |
Human serum 4 | 2.81 | 0.0337 | 1.2 |
Human serum 5 | 5.08 | 0.0540 | 1.1 |
Haptoglobin values for human serum and plasma samples obtained on a cobas c 503 analyzer (y) were compared with those determined using the corresponding reagent on a cobas c 501 analyzer (x).
Passing/Bablok LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790. | Linear regression |
y = 1.012x + 0.007 g/L | y = 0.971x + 0.0462 g/L |
τ = 0.989 | r = 0.999 |
The sample concentrations were between 0.110 and 5.62 g/L.
Haptoglobin values for human serum and plasma samples obtained on a cobas c 303 analyzer (y) were compared with those determined using the corresponding reagent on a cobas c 501 analyzer (x).
Passing/Bablok LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790. | Linear regression |
y = 1.014x + 0.0161 g/L | y = 0.969x + 0.0586 g/L |
τ = 0.977 | r = 0.999 |
The sample concentrations were between 0.150 and 5.60 g/L.
Haptoglobin is a transport and acute phase protein which is synthesized in hepatocytes. It is a glycoprotein which consists of two light α‑chains and two heavy β‑chains. The genetic polymorphism of the α‑chains leads to three phenotypes Hp 1‑1, Hp 2‑1 and Hp 2‑2 differing in molecular weight.
Haptoglobin binds hemoglobin in a strong haptoglobin‑hemoglobin complex (Hp‑Hb), the hemoglobin resulting from pathologically elevated hemolysis. These complexes are deposited in the hepatocytes, the deposition process having a half-life of less than 10 minutes. Hemoglobin is enzymatically metabolized and haptoglobin is liberated after approximately 3 days. Complex formation and the extremely rapid elimination from circulating blood prevent the occurrence of hemoglobinuria with excess renal loss of iron. A reduction in the level of free haptoglobin is indicative of intravascular hemolysis.
As a strong positive acute phase reactant, a hemolysis-mediated reduction or, to a certain extent, an elevation with accompanying acute inflammation can be compensated for. Indications for haptoglobin assays have been published and include the assessment of the severity and stage of intravascular hemolysis, evaluation of acute inflammatory processes.
Various methods including nephelometry, radial immunodiffusion (RID) and turbidimetric methods are available for the determination of haptoglobin. The haptoglobin assay from Roche is based on the principle of immunological agglutination.
R1 | Phosphate buffer: 12.7 mmol/L, pH 7.2; NaCl: 130 mmol/L; PEG: 40 g/L; preservative |
R3 | Anti‑human haptoglobin antibody (rabbit): > 1.1 g/L; NaCl: 100 mmol/L; preservative |
R1 is in position B and R3 is in position C.
For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.
Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.
Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.
Safety data sheet available for professional user on request.
For quality control, use control materials as listed in the “Order information” section. In addition, other suitable control material can be used.
The control intervals and limits should be adapted to each laboratory’s individual requirements. It is recommended to perform quality control always after lot calibration and subsequently at least every 26 weeks. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.
Follow the applicable government regulations and local guidelines for quality control.
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Serum
Plasma: Li‑heparin and K2‑EDTA plasma
The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.
Centrifuge samples containing precipitates before performing the assay.
See the limitations and interferences section for details about possible sample interferences.
Stability: LREFTöpfer G, Hornig F, Sauer K, et al. Untersuchungen zur Stabilität von 11 Serumproteinen bei Bestimmung mittels Immunturbidimetrie. J Lab Med 2000;24(3):118-125. | 3 months at 15‑25 °C 8 months at 2‑8 °C |
Sample stability claims were established by experimental data by the manufacturer or based on reference literature and only for the temperatures/time frames as stated in the method sheet. It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory.
In vitro test for the quantitative determination of haptoglobin in human serum and plasma on Roche/Hitachi cobas c systems.
Immunoturbidimetric assay.
Human haptoglobin forms a precipitate with a specific antiserum which is determined turbidimetrically.
Measuring range
0.1‑5.7 g/L (1.0‑57 µmol/L, 10‑570 mg/dL)
Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.
Lower limits of measurement
Lower detection limit of the test
0.1 g/L (1.0 µmol/L, 10 mg/dL)
The lower detection limit represents the lowest measurable analyte level that can be distinguished from zero. It is calculated as the value lying 3 standard deviations above that of the lowest standard (standard 1 + 3 SD, repeatability, n = 21).
0.3‑2.0 g/L (3.0‑20.0 µmol/L, 30‑200 mg/dL)
Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.
Criterion: Recovery within ± 10 % of the initial value at a haptoglobin concentration of 0.3 g/L (3.0 µmol/L, 30 mg/dL).
Icterus:
Hemolysis:
The Glick model which is normally used for assessment of hemoglobin interference is not suitable in the case of haptoglobin. Binding of free hemoglobin is the physiological function of haptoglobin. In the Glick study, hemolysate is added to the sample resulting in the formation of the haptoglobin-hemoglobin complex. This complex is present in the reagent tube and causes a 10‑15 % decrease in haptoglobin values. However, the effect is of no relevance for the results in native samples because in vivo the haptoglobin-hemoglobin complex is rapidly eliminated from the circulation and is practically not present in the blood.
Lipemia (Intralipid):
Rheumatoid factors up to 250 IU/mL do not interfere.
High dose hook-effect: No false result occurs up to a haptoglobin concentration of 12 g/L (120 µmol/L, 1200 mg/dL).
Drugs: No interference was found at therapeutic concentrations using common drug panels.
In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.
For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.
ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on cobas c systems. The latest version of the carry‑over evasion list can be found with the NaOHD-SMS-SmpCln1+2-SCCS Method Sheets. For further instructions refer to the operator’s manual. cobas c 502 analyzer: All special wash programming necessary for avoiding carry‑over is available via the cobas link, manual input is required in certain cases.
Where required, special wash/carry‑over evasion programming must be implemented prior to reporting results with this test.
|
| Analyzer(s) on which cobas c pack(s) can be used | |
03005593 322 | Tina-quant Haptoglobin ver.2, 100 tests | System-ID 07 9009 5 | cobas c 311, cobas c 501/502 |
Materials required (but not provided):
11355279 216 | Calibrator f.a.s. Proteins (5 × 1 mL) | Code 656 | |
11355279 160 | Calibrator f.a.s. Proteins (5 × 1 mL, for USA) | Code 656 | |
10557897 122 | Precinorm Protein 3 x 1 mL | Code 302 | |
10557897 160 | Precinorm Protein (3 × 1 mL, for USA) | Code 302 | |
11333127 122 | Precipath Protein (3 × 1 mL) | Code 303 | |
11333127 160 | Precipath Protein (3 × 1 mL, for USA) | Code 303 | |
05117003 190 | PreciControl ClinChem Multi 1 (20 x 5 mL) | Code 391 | |
05947626 190 | PreciControl ClinChem Multi 1 (4 x 5 mL) | Code 391 | |
05947626 160 | PreciControl ClinChem Multi 1 (4 x 5 mL, for USA) | Code 391 | |
05117216 190 | PreciControl ClinChem Multi 2 (20 x 5 mL) | Code 392 | |
05947774 190 | PreciControl ClinChem Multi 2 (4 x 5 mL) | Code 392 | |
05947774 160 | PreciControl ClinChem Multi 2 (4 x 5 mL, for USA) | Code 392 | |
04489357 190 | Diluent NaCl 9 % (50 mL) | System-ID 07 6869 3 |
For cobas c 311/501 analyzers:
HAPT2: ACN 228
For cobas c 502 analyzer:
HAPT2: ACN 8228
Ready for use
cobas c 311 test definition | |||
Assay type | 2‑Point End | ||
Reaction time / Assay points | 10 / 6-24 | ||
Wavelength (sub/main) | 700/340 nm | ||
Reaction direction | Increase | ||
Units | g/L (µmol/L, mg/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 110 µL | – | |
R2 | 50 µL | – | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (NaCl) | ||
Normal | 5.5 µL | 9 µL | 180 µL |
Decreased | 5.5 µL | 4 µL | 164 µL |
Increased | 5.5 µL | 9 µL | 180 µL |
cobas c 501 test definition | |||
Assay type | 2‑Point End | ||
Reaction time / Assay points | 10 / 10-48 | ||
Wavelength (sub/main) | 700/340 nm | ||
Reaction direction | Increase | ||
Units | g/L (µmol/L, mg/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 110 µL | – | |
R2 | 50 µL | – | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (NaCl) | ||
Normal | 5.5 µL | 9 µL | 180 µL |
Decreased | 5.5 µL | 4 µL | 164 µL |
Increased | 5.5 µL | 9 µL | 180 µL |
cobas c 502 test definition | |||
Assay type | 2‑Point End | ||
Reaction time / Assay points | 10 / 10-48 | ||
Wavelength (sub/main) | 700/340 nm | ||
Reaction direction | Increase | ||
Units | g/L (µmol/L, mg/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 110 µL | – | |
R2 | 50 µL | – | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (NaCl) | ||
Normal | 5.5 µL | 9 µL | 180 µL |
Decreased | 5.5 µL | 4 µL | 164 µL |
Increased | 5.5 µL | 18 µL | 180 µL |
HAPT2 | |
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label. |
On-board in use and refrigerated on the analyzer: | 12 weeks |
Diluent NaCl 9 % | |
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label. |
On-board in use and refrigerated on the analyzer: | 12 weeks |
Calibrators | S1: H2O S2-S6: C.f.a.s. Proteins | |
Multiply the lot-specific C.f.a.s. Proteins calibrator value by the factors below to determine the standard concentrations for the 6‑point calibration curve: | ||
S2: 0.0955 | S5: 1.45 | |
S3: 0.382 | S6: 2.28 | |
S4: 0.840 | ||
Calibration mode | RCM2 | |
Calibration frequency | Full calibration - after reagent lot change - as required following quality control procedures |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against the certified reference material in human serum of the IRMM (Institute for Reference Materials and Measurements) ERM‑DA470k/IFCC.
Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.
Precision was determined using human samples and controls in an internal protocol with repeatability (n = 21) and intermediate precision (3 aliquots per run, 1 run per day, 21 days). The following results were obtained:
Repeatability | Mean g/L | SD g/L | CV % |
Precinorm Protein | 1.05 (10.5, 105) | 0.01 (0.1, 1) | 0.7 |
Precipath Protein | 1.75 (17.5, 175) | 0.01 (0.1, 1) | 0.7 |
Human serum 1 | 1.03 (10.3, 103) | 0.00 (0.0, 0) | 0.4 |
Human serum 2 | 1.40 (14.0, 140) | 0.02 (0.2, 2) | 1.3 |
Intermediate precision | Mean g/L | SD g/L | CV % |
Precinorm Protein | 1.04 (10.4, 104) | 0.01 (0.1, 1) | 1.2 |
Precipath Protein | 1.73 (17.3, 173) | 0.02 (0.2, 2) | 1.1 |
Human serum 3 | 1.05 (10.5, 105) | 0.01 (0.1, 1) | 1.2 |
Human serum 4 | 1.57 (15.7, 157) | 0.02 (0.2, 2) | 1.2 |
Haptoglobin values for human serum and plasma samples obtained on a
Sample size (n) = 304
Passing/Bablok LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790. | Linear regression |
y = 0.996x + 0.014 g/L | y = 0.998x + 0.011 g/L |
τ = 0.974 | r = 0.999 |
The sample concentrations were between 0.030 and 5.32 g/L (0.300 and 53.2 µmol/L, 3.00 and 532 mg/dL). |
Haptoglobin is a transport and acute phase protein which is synthesized in hepatocytes. It is a glycoprotein which consists of two light α‑chains and two heavy β‑chains. The genetic polymorphism of the α‑chains leads to three phenotypes Hp 1‑1, Hp 2‑1 and Hp 2‑2 differing in molecular weight.
Haptoglobin binds hemoglobin in a strong haptoglobin‑hemoglobin complex (Hp‑Hb), the hemoglobin resulting from pathologically elevated hemolysis. These complexes are deposited in the hepatocytes, the deposition process having a half-life of less than 10 minutes. Hemoglobin is enzymatically metabolized and haptoglobin is liberated after approximately 3 days. Complex formation and the extremely rapid elimination from circulating blood prevent the occurrence of hemoglobinuria with excess renal loss of iron. A reduction in the level of free haptoglobin is indicative of intravascular hemolysis.
As a strong positive acute phase reactant, a hemolysis-mediated reduction or, to a certain extent, an elevation with accompanying acute inflammation can be compensated for. Indications for haptoglobin assays have been published and include the assessment of the severity and stage of intravascular hemolysis,
Various methods including nephelometry, radial immunodiffusion (RID) and turbidimetric methods are available for the determination of haptoglobin. The haptoglobin assay from Roche is based on the principle of immunological agglutination.
R1 | Phosphate buffer: 12.7 mmol/L, pH 7.2; NaCl: 130 mmol/L; PEG: 40 g/L; preservative |
R2 | Anti‑human haptoglobin antibody (rabbit): > 1.1 g/L; NaCl: 100 mmol/L; preservative |
R1 is in position B and R2 is in position C.
For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.
Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.
Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.
Safety data sheet available for professional user on request.
For USA: Caution: Federal law restricts this device to sale by or on the order of a physician.
For quality control, use control materials as listed in the \"Order information\" section.
In addition, other suitable control material can be used.
The control intervals and limits should be adapted to each laboratory’s individual requirements. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.
Follow the applicable government regulations and local guidelines for quality control.
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin and K2‑EDTA plasma
The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.
Centrifuge samples containing precipitates before performing the assay.
See the limitations and interferences section for details about possible sample interferences.
Stability: LREFTöpfer G, Hornig F, Sauer K, et al. Untersuchungen zur Stabilität von 11 Serumproteinen bei Bestimmung mittels Immunturbidimetrie. J Lab Med 2000;24(3):118-125. | 3 months at 15‑25 °C |
8 months at 2‑8 °C |
Sample stability claims were established by experimental data by the manufacturer or based on reference literature and only for the temperatures/time frames as stated in the method sheet. It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory.
In vitro test for the quantitative immunological determination of human haptoglobin in serum and plasma on COBAS INTEGRA systems.
Immunoturbidimetric assay
Human haptoglobin forms a precipitate with a specific antiserum which is determined turbidimetrically at 340 nm.
0.1‑5.14 g/L (1.00‑51.4 µmol/L or 10.0‑514 mg/dL) (typical measuring range)
The upper limit of the measuring range depends on the actual calibrator value.
Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:3 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 3.
Lower detection limit of the test:
0.1 g/L (1.00 µmol/L or 10.0 mg/dL)
The lower detection limit represents the lowest measurable analyte level that can be distinguished from zero. It is calculated as the value lying 3 standard deviations above that of a zero sample (zero sample + 3 SD, repeatability, n = 21).
0.3‑2.0 g/L (3.00‑20.0 µmol/L or 30‑200 mg/dL)
Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.
Criterion: Recovery within ± 10 % of initial value.
Serum, plasma
Icterus:
Hemolysis: The Glick model which is normally used for assessment of hemoglobin interference is not suitable in the case of haptoglobin. Binding of free hemoglobin is the physiological function of haptoglobin. In the Glick study, hemolysate is added to the sample resulting in the formation of the haptoglobin-hemoglobin complex. This complex is present in the reagent tube and causes a 10‑15 % decrease in haptoglobin values. However, the effect is of no relevance for the results in native samples because in vivo the haptoglobin-hemoglobin complex is rapidly eliminated from the circulation and is practically not present in the blood.
Lipemia (Intralipid):
Rheumatoid factors: No significant interference from rheumatoid factors up to a concentration of 1200 IU/mL.
Drugs: No interference was found at therapeutic concentrations using common drug panels.
In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.
For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.
ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on COBAS INTEGRA analyzers. Refer to the CLEAN Method Sheet for further instructions and for the latest version of the Extra wash cycle list.
Where required, special wash/carry-over evasion programming must be implemented prior to reporting results with this test.
|
| Analyzer(s) on which cobas c pack(s) can be used | |
03005593 322 | Tina-quant Haptoglobin ver.2 (100 tests) | System-ID 07 9009 5 | COBAS INTEGRA 400 plus |
Materials required (but not provided):
11355279 216 | C.f.a.s. Proteins (5 × 1 mL) | System-ID 07 6557 0 | |
11355279 160 | C.f.a.s. Proteins (5 × 1 mL, for USA) | System-ID 07 6557 0 | |
10557897 122 | Precinorm Protein (3 × 1 mL) | System-ID 07 9105 9 | |
10557897 160 | Precinorm Protein (3 × 1 mL, for USA) | System-ID 07 9105 9 | |
11333127 122 | Precipath Protein (3 × 1 mL) | System-ID 07 9106 7 | |
11333127 160 | Precipath Protein (3 × 1 mL, for USA) | System-ID 07 9106 7 | |
05117003 190 | PreciControl ClinChem Multi 1 (20 × 5 mL) | System-ID 07 7469 3 | |
05947626 190 | PreciControl ClinChem Multi 1 (4 × 5 mL) | System-ID 07 7469 3 | |
05947626 160 | PreciControl ClinChem Multi 1 (4 × 5 mL, for USA) | System-ID 07 7469 3 | |
05117216 190 | PreciControl ClinChem Multi 2 (20 × 5 mL) | System-ID 07 7470 7 | |
05947774 190 | PreciControl ClinChem Multi 2 (4 × 5 mL) | System-ID 07 7470 7 | |
05947774 160 | PreciControl ClinChem Multi 2 (4 × 5 mL, for USA) | System-ID 07 7470 7 | |
20756350 322 | NaCl Diluent 9 % (6 × 22 mL) | System-ID 07 5635 0 |
Test HAPT2, test ID 0‑009
Ready for use
Measuring mode | Absorbance |
Abs. calculation mode | Endpoint |
Reaction mode | D-R1-S-SR |
Reaction direction | Increase |
Wavelength A/B | 340 nm |
Calc. first/last | 33/55 |
Typical prozone effect | > 14.0 g/L (> 140 µmol/L or > 1400 mg/dL) |
Antigen excess check | No |
Predilution factor | 21 |
Unit | g/L |
Diluent (H2O) | ||
R1 | 100 µL | |
Sample | 5.5 µL | 14.5 µL |
SR | 50 µL | |
Total volume | 170 µL |
Shelf life at 2‑8 °C | See expiration date on cobas c pack label |
On-board in use at 10‑15 °C | 12 weeks |
Calibrator | C.f.a.s. Proteins |
Calibration dilution ratio | 1:9.2, 1:14.5, 1:25, 1:55, 1:220, performed automatically by the instrument, system water as zero calibrator |
Calibration mode | Logit/log 5 |
Calibration replicate | Duplicate recommended |
Calibration interval | Each lot and as required following quality control procedures |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Enter the assigned lot-specific haptoglobin value of the undiluted calibrator, indicated in the package insert of the C.f.a.s. Proteins.
Traceability: This method has been standardized against the certified reference material in human serum of the IRMM (Institute for Reference Materials and Measurements) ERM‑DA470k/IFCC.
Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.
Precision was determined using human samples and controls in an internal protocol with repeatability (n = 21) and intermediate precision (3 aliquots per run, 1 run per day, 10 days). The following results were obtained:
Repeatability | Mean | SD | CV |
|---|---|---|---|
Precinorm Protein | 1.12 (11.2, 112) | 0.01 (0.1, 1) | 1.0 |
Precipath Protein | 2.19 (21.9, 219) | 0.01 (0.1, 1) | 0.4 |
Human serum 1 | 0.794 (7.94, 79.4) | 0.010 (0.10, 1.0) | 1.2 |
Human serum 2 | 3.78 (37.8, 378) | 0.02 (0.2, 2) | 0.6 |
Intermediate precision | Mean | SD | CV |
|---|---|---|---|
Precinorm Protein | 1.06 (10.6, 106) | 0.01 (0.1, 1) | 1.0 |
Precipath Protein | 2.13 (21.3, 213) | 0.02 (0.2, 2) | 0.7 |
Human serum 1 | 0.812 (8.12, 81.2) | 0.012 (0.12, 1.2) | 3.0 |
Human serum 2 | 3.64 (36.4, 364) | 0.02 (0.2, 2) | 3.9 |
Haptoglobin values for human serum and plasma samples obtained on a COBAS INTEGRA 400 analyzer using the COBAS INTEGRA Tina-quant Haptoglobin ver.2 reagent (y) were compared with those determined using the same reagent on a COBAS INTEGRA 700 analyzer (x) and with those determined on a commercially available alternative automated system (nephelometric determination) (x).
COBAS INTEGRA 700 analyzer | |
Sample size (n) | 73 |
Correlation coefficient (r) | 0.999 |
Lin. regression | y = 0.99 x + 0.02 g/L |
Passing/Bablok LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790. | y = 0.99 x + 0.02 g/L |
Alternative system | |
Sample size (n) | 60 |
Correlation coefficient (r) | 0.989 |
Lin. regression | y = 0.82 x + 0.06 g/L |
Passing/Bablok LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790. | y = 0.83 x + 0.03 g/L |
The sample concentrations were between 0.1 and 4.1 g/L (1.00 and 41.0 µmol/L or 10.0 and 410 mg/dL).
Haptoglobin is a transport and acute phase protein which is synthesized in hepatocytes. It is a glycoprotein which consists of two light α‑chains and two heavy β‑chains. The genetic polymorphism of the α‑chains leads to three phenotypes Hp 1‑1, Hp 2‑1 and Hp 2‑2 differing in molecular weight.
Haptoglobin binds hemoglobin in a strong haptoglobin-hemoglobin complex (Hp‑Hb), the hemoglobin resulting from pathologically elevated hemolysis. These complexes are deposited in the hepatocytes, the deposition process having a half-life of less than 10 minutes. Hemoglobin is enzymatically metabolized and haptoglobin is liberated after approximately 3 days. Complex formation and the extremely rapid elimination from circulating blood prevents the occurrence of hemoglobinuria with excess loss of iron. A reduction in the level of free haptoglobin is indicative of intravascular hemolysis.
As a strong positive acute phase reactant, a hemolysis mediated reduction or, to a certain extent, an elevation with accompanying acute inflammation can be compensated for. Indications for haptoglobin assays have been published and include the assessment of the severity and stage of intravascular hemolysis,
Various methods including nephelometry, radial immunodiffusion (RID) and turbidimetric methods are available for the determination of haptoglobin. The haptoglobin assay from Roche is based on the principle of immunological agglutination.
R1 | Phosphate buffer: 12.7 mmol/L, pH 7.2; NaCl: 130 mmol/L; PEG: 40 g/L; preservative |
SR | Anti-human haptoglobin antibody (rabbit) > 1.1 g/L; NaCl: 100 mmol/L; preservative |
R1 is in position B and SR is in position C.
For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.
Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.
Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.
Safety data sheet available for professional user on request.
For USA: Caution: Federal law restricts this device to sale by or on the order of a physician.
Reference range | Precinorm Protein or PreciControl ClinChem Multi 1 |
Pathological range | Precipath Protein or PreciControl ClinChem Multi 2 |
Control interval | 24 hours recommended |
Control sequence | User defined |
Control after calibration | Recommended |
For quality control, use control materials as listed in the “Order information” section. In addition, other suitable control material can be used.
The control intervals and limits should be adapted to each laboratory’s individual requirements. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.
Follow the applicable government regulations and local guidelines for quality control.
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Serum
Plasma: Heparin (Li‑, Na‑, NH4+‑) or EDTA (K2‑, K3‑) plasma
The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.
Centrifuge samples containing precipitates before performing the assay.
Samples and controls are automatically prediluted 1:21 (1+20) with NaCl solution by the instrument.
See the limitations and interferences section for details about possible sample interferences.
Stability: LREFTöpfer G, Hornig F, Sauer K, et al. Untersuchungen zur Stabilität von 11 Serumproteinen bei Bestimmung mittels Immunturbidimetrie. J Lab Med 2000;24(3):118-125. | 3 months at 15‑25 °C 8 months at 2‑8 °C |
Sample stability claims were established by experimental data by the manufacturer or based on reference literature and only for the temperatures/time frames as stated in the method sheet. It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory.
Tina-quant Haptoglobin ver.2
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