Assays for viral sequences and their value in validation of viral elimination

Dev Biol Stand. 1993;81:231-6.


Plasma pools and blood products were analysed for hepatitis C virus (HCV) RNA by the polymerase chain reaction (PCR). Initially, samples were amplified by "nested" PCR using two pairs of primers from the highly-conserved 5' non-coding region of the genome. The method was later modified to amplification with only the outer pair of primers and a "hot-start" to eliminate amplification of non-target sequences. The modified method was as sensitive as "nested" PCR. However, with samples containing low levels of RNA, it is necessary to perform repeat amplifications to avoid false-negative results. Ten out of 17 anti-HCV positive pools from paid donors were positive, while none of the 12% pools from unpaid donors was positive for RNA or anti-HCV antibodies. Four i.v. immunoglobulins made from contaminated pools were virus-free, indicating removal of virus during current manufacturing processes. A quantitative PCR method was established using a positive donation. Probit analysis gave an estimated viral titre of 3.6 x 10(6) genomes/ml.

Publication types

  • Comparative Study

MeSH terms

  • Base Sequence
  • Biological Products / standards*
  • Blood / microbiology
  • Blood Donors
  • Drug Contamination / prevention & control
  • False Negative Reactions
  • Genome, Viral*
  • Hepacivirus / genetics
  • Hepacivirus / isolation & purification*
  • Hepatitis C / prevention & control
  • Hepatitis C / transmission
  • Humans
  • Immunoglobulins, Intravenous / isolation & purification
  • Mass Screening / methods*
  • Molecular Sequence Data
  • Polymerase Chain Reaction*
  • Quality Control
  • RNA, Viral / blood*
  • Sensitivity and Specificity
  • Volunteers


  • Biological Products
  • Immunoglobulins, Intravenous
  • RNA, Viral