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. 1993 Feb;31(2):289–291. doi: 10.1128/jcm.31.2.289-291.1993

Detection of hepatitis C virus RNA: comparison of one-stage polymerase chain reaction (PCR) with nested-set PCR.

D R Gretch 1, J J Wilson 1, R L Carithers Jr 1, C dela Rosa 1, J H Han 1, L Corey 1
PMCID: PMC262751  PMID: 8381810

Abstract

We evaluated a new hepatitis C virus RNA assay based on one-stage PCR followed by liquid hybridization with an oligonucleotide probe and compared it with nested-set PCR. The one-stage and nested-set PCR assays had identical sensitivities in analytical experiments and showed 100% concordance when clinical specimens were used. One-stage PCR may be less prone to contamination than nested-set PCR.

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Selected References

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  1. Brillanti S., Garson J. A., Tuke P. W., Ring C., Briggs M., Masci C., Miglioli M., Barbara L., Tedder R. S. Effect of alpha-interferon therapy on hepatitis C viraemia in community-acquired chronic non-A, non-B hepatitis: a quantitative polymerase chain reaction study. J Med Virol. 1991 Jun;34(2):136–141. doi: 10.1002/jmv.1890340213. [DOI] [PubMed] [Google Scholar]
  2. Bukh J., Purcell R. H., Miller R. H. Importance of primer selection for the detection of hepatitis C virus RNA with the polymerase chain reaction assay. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):187–191. doi: 10.1073/pnas.89.1.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cesarone C. F., Bolognesi C., Santi L. Improved microfluorometric DNA determination in biological material using 33258 Hoechst. Anal Biochem. 1979 Nov 15;100(1):188–197. doi: 10.1016/0003-2697(79)90131-3. [DOI] [PubMed] [Google Scholar]
  4. Cristiano K., Di Bisceglie A. M., Hoofnagle J. H., Feinstone S. M. Hepatitis C viral RNA in serum of patients with chronic non-A, non-B hepatitis: detection by the polymerase chain reaction using multiple primer sets. Hepatology. 1991 Jul;14(1):51–55. doi: 10.1002/hep.1840140109. [DOI] [PubMed] [Google Scholar]
  5. Farci P., Alter H. J., Wong D., Miller R. H., Shih J. W., Jett B., Purcell R. H. A long-term study of hepatitis C virus replication in non-A, non-B hepatitis. N Engl J Med. 1991 Jul 11;325(2):98–104. doi: 10.1056/NEJM199107113250205. [DOI] [PubMed] [Google Scholar]
  6. Garson J. A., Tedder R. S., Briggs M., Tuke P., Glazebrook J. A., Trute A., Parker D., Barbara J. A., Contreras M., Aloysius S. Detection of hepatitis C viral sequences in blood donations by "nested" polymerase chain reaction and prediction of infectivity. Lancet. 1990 Jun 16;335(8703):1419–1422. doi: 10.1016/0140-6736(90)91446-h. [DOI] [PubMed] [Google Scholar]
  7. Gretch D., Lee W., Corey L. Use of aminotransferase, hepatitis C antibody, and hepatitis C polymerase chain reaction RNA assays to establish the diagnosis of hepatitis C virus infection in a diagnostic virology laboratory. J Clin Microbiol. 1992 Aug;30(8):2145–2149. doi: 10.1128/jcm.30.8.2145-2149.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Han J. H., Shyamala V., Richman K. H., Brauer M. J., Irvine B., Urdea M. S., Tekamp-Olson P., Kuo G., Choo Q. L., Houghton M. Characterization of the terminal regions of hepatitis C viral RNA: identification of conserved sequences in the 5' untranslated region and poly(A) tails at the 3' end. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1711–1715. doi: 10.1073/pnas.88.5.1711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Inchauspe G., Abe K., Zebedee S., Nasoff M., Prince A. M. Use of conserved sequences from hepatitis C virus for the detection of viral RNA in infected sera by polymerase chain reaction. Hepatology. 1991 Oct;14(4 Pt 1):595–600. doi: 10.1016/0270-9139(91)90044-v. [DOI] [PubMed] [Google Scholar]
  10. Okamoto H., Okada S., Sugiyama Y., Yotsumoto S., Tanaka T., Yoshizawa H., Tsuda F., Miyakawa Y., Mayumi M. The 5'-terminal sequence of the hepatitis C virus genome. Jpn J Exp Med. 1990 Jun;60(3):167–177. [PubMed] [Google Scholar]
  11. Schlauder G. G., Leverenz G. J., Mattsson L., Weiland O., Mushahwar I. K. Detection of hepatitis C viral RNA by the polymerase chain reaction in serum of patients with post-transfusion non-A, non-B hepatitis. J Virol Methods. 1992 May;37(2):189–199. doi: 10.1016/0166-0934(92)90046-g. [DOI] [PubMed] [Google Scholar]
  12. Shindo M., Di Bisceglie A. M., Cheung L., Shih J. W., Cristiano K., Feinstone S. M., Hoofnagle J. H. Decrease in serum hepatitis C viral RNA during alpha-interferon therapy for chronic hepatitis C. Ann Intern Med. 1991 Nov 1;115(9):700–704. doi: 10.7326/0003-4819-115-9-700. [DOI] [PubMed] [Google Scholar]
  13. Simmonds P., Zhang L. Q., Watson H. G., Rebus S., Ferguson E. D., Balfe P., Leadbetter G. H., Yap P. L., Peutherer J. F., Ludlam C. A. Hepatitis C quantification and sequencing in blood products, haemophiliacs, and drug users. Lancet. 1990 Dec 15;336(8729):1469–1472. doi: 10.1016/0140-6736(90)93179-s. [DOI] [PubMed] [Google Scholar]
  14. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  15. Tait J. F., Frankenberry D. A., Miao C. H., Killary A. M., Adler D. A., Disteche C. M. Chromosomal localization of the human annexin III (ANX3) gene. Genomics. 1991 Jun;10(2):441–448. doi: 10.1016/0888-7543(91)90330-h. [DOI] [PubMed] [Google Scholar]
  16. Ulrich P. P., Romeo J. M., Lane P. K., Kelly I., Daniel L. J., Vyas G. N. Detection, semiquantitation, and genetic variation in hepatitis C virus sequences amplified from the plasma of blood donors with elevated alanine aminotransferase. J Clin Invest. 1990 Nov;86(5):1609–1614. doi: 10.1172/JCI114882. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Weiner A. J., Kuo G., Bradley D. W., Bonino F., Saracco G., Lee C., Rosenblatt J., Choo Q. L., Houghton M. Detection of hepatitis C viral sequences in non-A, non-B hepatitis. Lancet. 1990 Jan 6;335(8680):1–3. doi: 10.1016/0140-6736(90)90134-q. [DOI] [PubMed] [Google Scholar]

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