Cobas ampliprep

Cobas Ampliprep-Cobas TaqMan is an automated RT-PCR test based on a dual-labeled hybridization probe targeting the precore and core regions associated with an HBV DNA automated extraction based on the affinity of DNA for silica gel-covered magnetic beads. An internal quantitation standard (QS) is added to each sample during the processing step. After HBV DNA extraction with the Cobas Ampliprep instrument, an RT-PCR test is performed by the CTM 48 analyzer with a multiplex TaqMan assay. Two targets, HBV DNA and the internal QS, are amplified.
For ensuring transfusion safety, blood units positive with either of the two tests or both the tests, were marked “positive” and discarded. The algorithm of the present study is shown in Figure 1.

Eligible PWH with a positive HBsAg on rapid test (Core Diagnostics, Birmingham, UK) were enrolled, and scheduled for a study visit. At the visit, demographics, clinical charateristics, existing CD4⁺ cell counts, and HIV viral loads were collected using standardized forms. Self-report adherence using the five-point Likert scale, which employs a 7-day recall, was administered to assess the level of adherence [21 (link)]. Blood samples were collected for laboratory confirmatory HBsAg status, renal and liver function tests. Also, blood samples were collected into two 4 ml EDTA vacutainers and plasma separated, aliquoted and stored for HBV DNA and HIV RNA quantification at the end of enrollment. Plasma HIV viral load was measured by real-time PCR using COBAS AmpliPrep and COBAS TaqMan Analyzer at the Metropolis Healthcare Limited Laboratory, an ISO certified commercial laboratory service provider in Ghana. The lower limit of detection of the TaqMan assay was 20 copies/ml and the linear range 20–10 000 000 copies/ml. HIV RNA of at least 20 copies/ml was considered unsuppressed. Plasma HBV DNA quantification was performed using fully automated COBAS TaqMan Analyzer (Roche Diagnostics GmbH, Mannheim, Germany). The lower limit of detection of the Roche TaqMan assay is 20 IU/ml and the linear range 20–170 000 000 IU/ml. HBV DNA of at least 20 IU/ml was considered unsuppressed.

Before beginning the antiviral treatment, the following variables were collected:
-demographic, anthropometric, and epidemiological: sex, age, body mass index (BMI), human immunodeficiency virus or hepatitis B virus co-infection, presence of diabetes mellitus.
-histological and other pathologies related to liver disease: degree of liver fibrosis (measured by transition electrographic image, FibroScan®), presence of cirrhosis, complications related to liver disease and extrahepatic manifestations.
-laboratory tests: leukocytes, haemoglobin, platelets, glucose, creatinine, albumin, total cholesterol, aspartate-aminotransferase (AST), alanine-aminotransferase (ALT), gamma-glutamyl transferase (GGT), alkaline-phosphatase (ALP), total bilirubin.
-virological: HCV viral load (HCV-RNA amplification was carried out with COBAS-AmpliPrep equipment and the polymerase chain reaction with COBAS-Taqman), HCV genotype.
-pharmacotherapeutics: prescribed pharmacotherapeutics regimen (drugs and duration).
During treatment and until week 24 post-treatment, ADRs that occurred were recorded. In the 12 or 24 week post-treatment, achievement of SVR was recorded.