For Research Use Only. Not for use in diagnostic procedures. Others cobas HBV RNA Test 5800-6800-8800 RUO cobas® HBV RNA PID00000472 Quantitative nucleic acid test for use on the cobas® 5800/6800/8800 Systems 09 700 064 190 9 700 064 190 09700064190 9700064190 09700064190 KIT COBAS 58/68/88 HBV RNA 192T RUO KIT COBAS 58/68/88 HBV RNA 192T RUO 00875197007152 Reagents, kits 1 Kit 192 Tests true cobas® HBV RNA for use on the cobas® 5800/6800/8800 Systems (cobas® HBV RNA) is an automated real-time RT-PCR assay for the in vitro quantitative detection of circulating HBV RNA in EDTA plasma and serum. cobas® HBV RNA is intended for research use only and is not for use in diagnostic procedures. cobas® HBV RNA can be used to support scientific research to understand the potential use of this new biomarker. en The cobas® HBV RNA test 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 Systems software, which assigns test results for all tests. Results can be reviewed directly on the system screen, and printed as a report.Armored RNA quantitation standard (QS) molecules are added during sample preparation. Nucleic acids (from samples and QS) are released by addition of proteinase and lysis reagent to the sample during universal sample preparation steps. The released nucleic acids bind to the silica surface of the added magnetic glass particles. In addition, the test utilizes three external controls: a high titer positive, a low titer positive, and a negative control.Unbound substances and impurities, such as denatured proteins, cellular debris, and potential PCR inhibitors (such as hemoglobin) from the lysed sample are reduced with subsequent wash reagent steps and purified nucleic acids are eluted from the glass particles with elution buffer at elevated temperature.Selective amplification of target nucleic acid HBV RNA from the sample is achieved by the use of organism-specific forward and reverse primers, which were selected to specifically hybridize to highly conserved regions of the HBV pre-genomic nucleic acid. A thermostable DNA polymerase enzyme is used for both reverse-transcription and amplification. The master mix includes deoxyuridine triphosphate (dUTP), instead of deoxythimidin e triphosphate (dTTP), which is incorporated into the newly synthesized DNA (amplicon). Any contaminating amplicons from previous PCR runs are destroyed by the AmpErase enzyme [uracil-N-glycosylase], which is included in the PCR mix, when heated in the first thermal cycling step. However, newly formed amplicons are not destroyed since the AmpErase enzyme is inactivated once exposed to temperatures above 55°C.The cobas® HBV RNA master mix contains detection probes which are specific for HBV and QS nucleic acid. The specific HBV and QS detection probes are each labeled with one of two unique fluorescent dyes which act as a reporter. Each probe also has a second dye which acts as a quencher.The fluorescent signals of the intact probes are suppressed by the quencher dye. During the PCR amplification step, hybridization of the probes to the specific single-stranded DNA template 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. Since the two specific reporter dyes are measured at defined wavelengths, simultaneous detection and discrimination of the amplified HBV RNA target and the QS are possible. en

cobas^® HBV RNA

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

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