title: Simultaneous extraction of hepatitis C virus (HCV), hepatitis B virus, and HIV-1 from plasma and detection of HCV RNA by a reverse transcriptase-polymerase chain reaction assay designed for screening pooled units of donated blood.
abstract: BACKGROUND
  Testing for viral nucleic acids should reduce the residual risk of transmitting viral infections by transfusion of blood components. The AmpliScreen Hepatitis C (HCV) Test, Version 2.0, was designed for screening pools composed of samples from individual units of blood or plasma.
  
  
  METHODS
  An ultracentrifugation step during sample processing simultaneously extracts and concentrates HCV, HIV type 1, and hepatitis B virus particles from plasma. An HCV internal control RNA serves as an extraction and amplification control for each processed sample. Processed samples are amplified by reverse transcriptase polymerase chain reaction and detected by hybridization of the amplified products to HCV- and internal-control-specific oligonucleotide probes. Plasma samples containing known quantities of HCV were used to evaluate analytical sensitivity and precision.
  
  
  RESULTS
  The analytical sensitivity of the test was 25 IU of HCV per mL of pooled plasma; all HCV genotypes were detected with similar efficiency. The test did not react with other blood-borne viruses. The within-run and total coefficients of variation were 1.3-13.0 percent and 1.7-22.0 percent, respectively, with low copy samples producing the more variable results. The test performed well using plasma collected in either EDTA, ACD, or PPT Vacutainer tubes. Plasma samples containing elevated levels of hemoglobin, albumin, triglycerides, or bilirubin did not interfere with the test. The test detected HCV RNA 23 to 32 days prior to seroconversion for four of the five seroconversion panels tested.
  
  
  CONCLUSIONS
  The AmpliScreen HCV Test, Version 2.0 provides a reproducible and specific method for screening pooled blood units for HCV. Theoretically, this test has sufficient sensitivity to detect a single infected unit containing 2.4 x 10(3) IU of HCV per mL in a pool with 95 uninfected units and should reduce the window period by at least 20 to 30 days.
authors.affiliation: Roche Molecular Systems, Inc., Somerville, New Jersey 08876, USA.
authors.forename: R
authors.initials: R
authors.lastname: Sun
authors.forename: W
authors.initials: W
authors.lastname: Schilling
authors.forename: H
authors.initials: H
authors.lastname: Jayakar
authors.forename: J
authors.initials: J
authors.lastname: Ku
authors.forename: J
authors.initials: J
authors.lastname: Wang
authors.forename: M
authors.initials: M
authors.lastname: Rosenstraus
authors.forename: J
authors.initials: J
authors.lastname: Spadoro
mesh_terms: D001769:Blood / Q000821:virology*; D001782:Blood Donors; D001800:Blood Specimen Collection / Q000295:instrumentation; D018067:Flaviviridae / Q000302:isolation & purification*; D005838:Genotype; D015497:HIV-1 / Q000302:isolation & purification*; D016174:Hepacivirus / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D008403:Mass Screening / Q000379:methods / Q000592:standards; D012367:RNA, Viral / Q000097:blood*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D012680:Sensitivity and Specificity; D012698:Serologic Tests
chemical_list: D012367:RNA, Viral