Experience with PCR screening

Dev Biol (Basel). 2000:102:81-91.

Abstract

The COBAS AmpliScreen Hepatitis C (HCV) Test, Version 2.0, which is designed for screening pools composed of samples from individual units of blood or plasma, employs a MultiPrep sample processing procedure that simultaneously extracts and concentrates HCV, HIV-1 and Hepatitis B virus particles from plasma. An HCV Internal Control (IC) RNA serves as an extraction and amplification control for each independently processed sample. Processed samples are amplified by RT-PCR using HCV-specific complementary primers and detected by hybridization of the amplified products to HCV- and IC-specific oligonucleotide probes. The analytical sensitivity of the test is 25 International Units (IU) of HCV per mL of pooled plasma; all HCV genotypes are detected with similar efficiency. The test detected HCV RNA 23 to 32 days prior to anti-HCV antibody seroconversion for four of the five seroconversion panels tested. The test had sufficient sensitivity to reproducibly detect a single infected unit containing 2.4 x 10(3) copies of HCV per mL in a pool with 23 uninfected units. COBAS AmpliScreen tests for HIV-1 and HBV now being validated by Roche Molecular Systems also incorporate the MultiPrep specimen processing method, thereby making it possible to use a single processed specimen to screen for all three viruses.

MeSH terms

  • Genotype
  • HIV Infections / blood
  • HIV Infections / virology
  • Hepacivirus / genetics
  • Hepacivirus / isolation & purification*
  • Hepatitis B / blood
  • Hepatitis B / virology
  • Hepatitis C / blood
  • Hepatitis C / prevention & control*
  • Hepatitis C / virology
  • Humans
  • Mass Screening / methods
  • Polymerase Chain Reaction / methods
  • RNA, Viral / blood*
  • Reagent Kits, Diagnostic / statistics & numerical data*
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • RNA, Viral
  • Reagent Kits, Diagnostic