title: Performance characteristics of the AmpliScreen HIV-1 test, an assay designed for screening plasma mini-pools.
abstract: This study evaluated the performance characteristics of the AmpliScreen(TM)Human Immunodeficiency Virus-Type 1 (HIV-1) Test, Version 1.5, a test designed for screening pools composed of samples from individual units of blood or plasma. HIV-1, hepatitis C (HCV) and hepatitis B (HBV) virus particles were simultaneously extracted and concentrated from plasma by a multi-prep sample processing procedure. An HIV-1 Internal Control (IC) RNA was added to each sample to serve as an extraction and amplification control. Processed samples were amplified by RT-PCR using HIV-1-specific complementary primers and detected by hybridization of the amplified products to HIV-1- and IC-specific oligonucleotide probes. The analytical sensitivity of the test (concentration that yields >/=95% positive results in a set of replicate tests) was 25 copies of HIV-1 RNA per mL of pooled plasma. Representative strains from all HIV-1 group M subtypes were reproducibly detected (>95% positive results among 22 replicate tests) at concentrations of 30 to 75 viral particles per ml. The test exhibited excellent specificity; it did not cross-react with a set of 30 viral and five bacterial isolates and yielded negative results on a panel of 500 blood samples from HIV-1 seronegative donors. Samples containing abnormally high levels of haemoglobin, albumin, triglycerides or bilirubin in plasma samples did not interfere with the detection of HIV-1 RNA at a concentration of 100 copies of per ml. The test detected HIV-1 RNA 7-17 days prior to anti-HIV-1 antibody seroconversion for all 10 seroconversion panels tested. A fully automated COBAS AmpliScreen(TM)version of this test is being validated. COBAS AmpliScreen tests for HCV and HBV also incorporate the multi-prep specimen processing method, thereby making it possible to use a single processed specimen to screen for all three viruses.
authors.affiliation: Roche Molecular Systems, Inc., Pleasanton, CA 94588, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.forename: M H
authors.initials: MH
authors.lastname: Wisbeski
authors.forename: M
authors.initials: M
authors.lastname: Mendoza
authors.forename: S
authors.initials: S
authors.lastname: Dorf
authors.forename: D
authors.initials: D
authors.lastname: Xu
authors.forename: M
authors.initials: M
authors.lastname: Nguyen
authors.forename: S
authors.initials: S
authors.lastname: Yeh
authors.forename: R
authors.initials: R
authors.lastname: Sun
mesh_terms: D005069:Evaluation Studies as Topic; D006679:HIV Seropositivity / Q000821:virology; D015497:HIV-1 / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008403:Mass Screening / Q000379:methods / Q000592:standards / Q000706:statistics & numerical data; D021141:Nucleic Acid Amplification Techniques; D010949:Plasma / Q000821:virology; D012367:RNA, Viral / Q000097:blood* / Q000235:genetics*; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral