Cobas taqman system

In total, we recruited 196 patients into our cohort, from different provinces of China, who were diagnosed in Qingchun Hospital of Zhejiang Province from January 2000 to January 2018, and received, or volunteered to rejected HAART. The viral loads of the patients were detected using the Cobas Taqman system (Roche, Basle, Switzerland). The healthy controls (HCs) were randomly selected from hospital clinics and were not suffering from any diseases. We chose two ECs, two HIV-positive infected patients (HPs), and two HCs of similar ages, sex, and traditional risk factors to perform second generation transcriptome sequencing. ECs were defined as HIV-infected patients with an undetectable viral load in the absence of HAART for nearly 10 years.

The Roche CMV assay relies on the Cobas AmpliPrep/Cobas^® TaqMan system, which consists of a Cobas AmpliPrep system for automatic nucleic acid isolation with magnetic beads and Cobas^® TaqMan system for real-time PCR (Roche Molecular Diagnostics, Pleasanton, CA, USA). In the current study, the Roche assay was only used for the analysis of plasma specimens, as per manufacturer’s recommendations.

A minimum of three recordings of liver biochemistry for patients with normal levels and two recordings for those with elevated levels were required. The median number of ALT tests performed pre liver biopsy was 5 (interquartile range [IQR] 4–8), and these were sampled over a median period of 21 months (IQR 8–46). Serum biochemical and haematological values prior to the liver biopsy were recorded. Routine liver biochemical tests were performed using commercially available autoanalysers and hepatitis serological markers were assayed using commercially available enzyme-linked immunoassays. Patients with HBV DNA <20,000 IU/mL were classified as low-replicative while those with HBV DNA ≥20,000 IU/mL were classified as high-replicative. A minimum of three HBV DNA recordings was required and the reference value utilized for analysis was based on the pre biopsy level. Serum HBV DNA level was expressed in IU/mL (1 IU/mL = 5.6 copies/mL) [7 (link)]. Quantitative HBV DNA levels were measured by a COBAS TaqMan System (Roche Diagnostics, Indianapolis, IN, USA), which has a lower detection limit of 15 IU/mL, or Abbott Real-Time HBV assay (Abbott Molecular, Inc., Des Plaines, IL, USA), with a lower detection limit of 10 IU/mL. The median duration between the first and pre liver biopsy HBV DNA was 12 months (IQR 6–20).