For in vitro diagnostic use. Others cobas BKV Test 5800-6800-8800 IVD cobas® BKV PID00000145 Quantitative nucleic acid test for use on the cobas® 5800/6800/8800 systems cobas® BKV is an in vitro nucleic acid amplification test for the quantitation of BK virus (BKV) DNA in human EDTA plasma and urine stabilized in cobas® PCR Media on the cobas® 5800/6800/8800 Systems.In EDTA plasma, cobas® BKV is intended for use as an aid in the management of BKV in transplant patients. In patients undergoing monitoring of BKV in EDTA plasma, serial DNA measurements can be used to indicate the need for potential treatment changes and to assess viral response to treatment.In urine stabilized in cobas® PCR Media, cobas® BKV is intended for use as an aid in the management of BKV in transplant patients.The results from cobas® BKV are intended to be read and analyzed by a qualified licensed healthcare professional in conjunction with clinical signs and symptoms and relevant laboratory findings. Test results must not be the sole basis for patient management decisions.cobas® BKV is not intended for use as a screening test for blood or blood products or human cells, tissues, and cellular and tissue-based products (HCT/Ps). en cobas® BKV is based on fully automated sample preparation (nucleic acid extraction and purification) followed by PCR amplification and detection. The cobas® 5800 system is designed as one integrated instrument. The cobas® 6800/8800 systems consist of the sample supply module, the transfer module, the processing module, and the analytic module. Automated data management is performed by the cobas® 5800 or cobas® 6800/8800 system software which assigns test results for all tests as either target not detected, BKV DNA detected < LLoQ (lower limit of quantitation), BKV DNA detected > ULoQ (upper limit of quantitation), or a value in the linear range LLoQ < x < ULoQ. Results can be reviewed directly on the system screen, exported, or printed as a report. Nucleic acid from patient samples and added lambda DNA-QS molecules is simultaneously extracted. In summary, viral nucleic acid is released by addition of proteinase and lysis reagent to the sample. The released nucleic acid binds to the silica surface of the added magnetic glass particles. Unbound substances and impurities, such as denatured protein, cellular debris and potential PCR inhibitors are removed with subsequent wash reagent steps and purified nucleic acid is eluted from the glass particles with elution buffer at elevated temperature. Selective amplification of target nucleic acid from the sample is achieved by the use of a dual target virus specific approach from highly-conserved regions of the BKV located in the BKV small t-antigen region and the BKV VP2 region. Selective amplification of DNA-QS is achieved by the use of sequence-specific forward and reverse primers which are selected to have no homology with the BKV genome. A thermostable DNA polymerase enzyme is used for amplification. The target and DNA-QS sequences are amplified simultaneously utilizing a universal PCR amplification profile with predefined temperature steps and number of cycles. The master mix includes deoxyuridine triphosphate (dUTP), instead of deoxythimidine triphosphate (dTTP), which is incorporated into the newly synthesized DNA (amplicon).10, 11,12 Any contaminating amplicon from previous PCR runs is eliminated by the AmpErase enzyme, which is included in the PCR mix, when heated in the first thermal cycling step. However, newly formed amplicon are not eliminated since the AmpErase enzyme is inactivated once exposed to temperatures above 55°C. The cobas® BKV master mix contains two detection probes specific for BKV target sequences and one for the DNA-QS. The probes are labeled with target-specific fluorescent reporter dyes allowing simultaneous detection of BKV target and DNA-QS in two different target channels.13,14 The fluorescent signal of the intact probes is suppressed by the quencher dye. During the PCR amplification step, hybridization of the probe to the specific single-stranded DNA templates results in cleavage by the 5'-to-3' nuclease activity of the DNA polymerase resulting in separation of the reporter and quencher dyes and the generation of a fluorescent signal. With each PCR cycle, increasing amounts of cleaved probes are generated and the cumulative signal of the reporter dye is concomitantly increased. Real-time detection and discrimination of PCR products are accomplished by measuring the fluorescence of the released reporter dyes for the viral targets and DNA-QS. 10. Longo MC, Berninger MS, Hartley JL. Use of uracil DNA glycosylase to control carry-over contamination in polymerase chain reactions. Gene. 1990;93:125-8. PMID: 2227421. 11. Savva R, McAuley-Hecht K, Brown T, Pearl L. The structural basis of specific base-excision repair by uracil-DNA glycosylase. Nature. 1995;373:487-93. PMID: 7845459. 12. Mol CD, Arvai AS, Slupphaug G, et al. Crystal structure and mutational analysis of human uracil-DNA glycosylase: structural basis for specificity and catalysis. Cell. 1995;80:869-78. PMID: 7697717. 13. Higuchi R, Dollinger G, Walsh PS, Griffith R. Simultaneous amplification and detection of specific DNA sequences. Biotechnology (N Y). 1992;10:413-7. PMID: 1368485. 14. Heid CA, Stevens J, Livak KJ, Williams PM. Real time quantitative PCR. Genome Res. 1996;6:986-94. PMID: 8908518. en

cobas® BKV

Quantitative nucleic acid test for use on the cobas^® 5800/6800/8800 systems

IVD

Setting a new standard for transplant patients

BK Polyomavirus (BKPyV) is a significant pathogen in transplant recipients, where viral reactivation can lead to serious complications such as polyomavirus-associated nephropathy (PVAN) and hemorrhagic cystitis, posing a direct threat to allograft survival and patient health^.1,2 Early detection with best-in-class monitoring tools provides actionable results, enabling informed treatment decisions.

The cobas® BKV test provides reliable, standardized, high-quality results for the improved management of transplant patients at risk of infection. The BKV test is intended for use as an aid in the management of BKV in kidney and hematopoietic stem cell transplant patients, with results used to indicate the need for potential treatment changes.³The test is for use on the automated cobas® 5800/6800/8800 systems.

Product specification	Value
Dual assay targets	VP2 and small t-Antigen
Sample types	EDTA Plasma Urine stabilized in cobas® PCR media
Minimum amount of sample required	EDTA Plasma: 350 µL* Urine: 550 µL**
Sample processing volume	EDTA Plasma: 200 µL Urine: 400 µL
Analytical sensitivity	EDTA Plasma: 21.5 IU/mL (95% CI 16.3 IU/mL - 32.4 IU/mL) Urine: 12.2 IU/mL (95% CI 9.2 IU/mL - 18.3 IU/mL)
Linear range	EDTA Plasma: 21.5 IU/mL to 1E+08 IU/mL Urine: 200 IU/mL to 1E+08 IU/mL
Overall precision	EDTA Plasma: SD 0.03 - 0.13 log₁₀ Urine: SD 0.02 - 0.07 log₁₀
Subtypes detected	Subtypes I (Ia, Ib, and Ic), II, III and IV

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Setting a new standard for transplant patients

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cobas® BKV test for cobas® 5800/6800/8800 systems performance³

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cobas® BKV

Setting a new standard for transplant patients

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Overview

Ordering information

Technical documents

cobas® BKV test for cobas® 5800/6800/8800 systems performance3

cobas® BKV test for cobas® 5800/6800/8800 systems performance3

Compatible products

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Benefits at a glance³

cobas® BKV test for cobas® 5800/6800/8800 systems performance³

cobas® BKV test for cobas® 5800/6800/8800 systems performance³