On investigation of the patient's immune status, it was established that she was HIV-1-positive as confirmed by HIV-1 DNA Amplicor
assay version 1.5 (Roche Diagnostics, Mannheim, Germany), with an HIV-1 viral load of 2 738 '30 RNA copies/ml by Abbott m2000 Real Time HIV 1 Assay (Abbott Diagnostics, Johannesburg) and a CD4+ lymphocyte percentage of 28%.
For all the subjects found positive for anti HCV antibody, ALT levels were done using kit by MERCK on the serum samples and the stored plasma was used for qualitative PCR for HCV RNA using the second generation 'Amplicor
HCV Assay by Roche Molecular Systems'.
Explain the methodology of the COBAS AMPLICOR
Detection and quantification of HCV-RNA in serum: Detection of HCV-RNA in serum was performed by automated RT-PCR assay (COBAS AMPLICOR
HCV Test, version 2.0; Roche Diagnostics, Molecular Division).
An aliquot of each sample was processed with the Amplicor
PCR test (Roche Molecular Systems, Branchburg, NJ, USA) for detection of C.
We used the Roche COBAS Amplicor
HBV Monitor test (Ver.
The currently available NAATs include PCR (COBAS Roche Amplicor
), strand displacement amplification (SDA) (BD ProbeTec), and transcription-mediated amplification (TMA) (Gen-Probe Aptima).
(10) There are currently 3 commercially available procedures for HCV viral load testing: the Roche (Belleville, NJ) AMPLICOR
manual and COBAS HCV assays; the Bayer (Norwood, Mass) QUANTIPLEX HCV RNA version 3.0 assay (branched DNA [bDNA]); and the Biomerieux (Marcy-l'Etole, France) NucliSens system, which uses nucleic acid sequence-based amplification.
Unfortunately, Roche Diagnostics has stopped making the Amplicor
EV that was used in this study.
Automated quantitative analysis of hepatitis B virus DNA by using the Cobas Amplicor
HBV monitor test.
Blood samples were collected for determination of [CD4.sup.+] T-cell counts (FACSCalibur system, Becton Dickinson, USA) and viral load tests (Roche Cobas Amplicor
HIV-1 RNA monitor version 1.5 assay, Roche Diagnostics, USA).
(9) Plasma HIV-1 RNA (viral load) was measured by polymerase chain reaction using the Amplicor
HIV-1 Monitor (Amplicor
Roche Diagnostics), which has a detection limit of 400 copies/mL; values were expressed as log (10).