{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,4]],"date-time":"2025-11-04T22:26:42Z","timestamp":1762295202508},"reference-count":29,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2005,12,7]],"date-time":"2005-12-07T00:00:00Z","timestamp":1133913600000},"content-version":"vor","delay-in-days":4784,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[1992,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>A procedure is described for the detection of hepatitis C virus (HCV) RNA in blood by means of the polymerase chain reaction (PCR) in which the reverse transcription step and two rounds of amplification are carried out in a single tube. This results in fewer manipulations, reduced risk of contamination, and economy of time. The procedures are generally applicable to other assays based on the PCR. We describe the preparation (from 100 \u03bcL serum) of test samples that remain stable for at least 6 days under specified conditions and an assay that employs nested primer pairs homologous to conserved sequences in the 5\u2032 noncoding region. The method was tested on 107 sera from the United States and Japan. Correlation with first\u2010generation anti\u2010HCV was 77%. Two sets of nested primer pairs homologous to sequences in the 5\u2032 noncoding region and one set based on structural region sequences showed differences in their reactivities with serum HCV RNA. The recommended single tube procedure specified a primer for reverse transcription that was conserved in all reported HCV genomes but absent from pestivirus genomic sequences. The effects of preanalytical factors on the detection of HCV RNA were studied. Qualitatively, there was no change in the HCV RNA\u2010positivity of sera that were exposed to room temperature for 24 hours. Quantitative studies showed a decrease in titer in some specimens. Three cycles of freeze\u2010thawing had no detectable effects on the titers of HCV RNA. \u00a9 1992 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/jmv.1890380312","type":"journal-article","created":{"date-parts":[[2007,3,2]],"date-time":"2007-03-02T21:32:54Z","timestamp":1172871174000},"page":"220-225","source":"Crossref","is-referenced-by-count":40,"title":["Polymerase chain reaction assay for hepatitis C virus RNA using a single tube for reverse transcription and serial rounds of amplification with nested primer pairs"],"prefix":"10.1002","volume":"38","author":[{"given":"Hsiang Ju","family":"Lin","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Naiyi","family":"Shi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masashi","family":"Mizokami","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"F. Blaine","family":"Hollinger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,12,7]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199111073251901"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.89.1.187"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(87)90021-2"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/hep.1840140109"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199107113250205"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(91)90269-H"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/0140-6736(90)92384-T"},{"key":"e_1_2_1_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/0140-6736(90)91446-H"},{"key":"e_1_2_1_10_1","first-page":"11","volume-title":"Polymerase Chain Reaction","author":"Gelfand DH","year":"1989"},{"key":"e_1_2_1_11_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.5.1711"},{"key":"e_1_2_1_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/0016-5085(92)90791-V"},{"key":"e_1_2_1_13_1","unstructured":"HoughtonM ChooQL KuoG(1988): Eur. Patent Applic. 88 310 922.5 and Publ. 0 318 216."},{"key":"e_1_2_1_14_1","first-page":"328","volume-title":"Viral Hepatitis and Liver Disease","author":"Houghton M","year":"1991"},{"key":"e_1_2_1_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/hep.1840140504"},{"key":"e_1_2_1_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/hep.1840140404"},{"key":"e_1_2_1_17_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.22.10292"},{"key":"e_1_2_1_18_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.87.24.9524"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/339237a0"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.8.3392"},{"key":"e_1_2_1_21_1","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-72-11-2697"},{"key":"e_1_2_1_22_1","first-page":"215","article-title":"Detection of hepatitis C virus RNA by a two\u2010stage polymerase chain reaction","volume":"60","author":"Okamoto H","year":"1990","journal-title":"Japanese Journal of Experimental Medicine"},{"key":"e_1_2_1_23_1","first-page":"167","article-title":"The 5\u2032 terminal sequences of the hepatitis C virus genome","volume":"60","author":"Okamoto H","year":"1990","journal-title":"Japanese Journal of Experimental Medicine"},{"key":"e_1_2_1_24_1","first-page":"25","volume-title":"Polymerase Chain Reaction","author":"Saiki R","year":"1989"},{"key":"e_1_2_1_25_1","first-page":"8","volume-title":"Molecular Cloning","author":"Sambrook J","year":"1989"},{"key":"e_1_2_1_26_1","doi-asserted-by":"crossref","first-page":"1105","DOI":"10.1128\/jvi.65.3.1105-1113.1991","article-title":"Structure and organization of the hepatitis C virus genome isolated from human carriers","volume":"65","author":"Takamizawa A","year":"1991","journal-title":"Journal of Virology"},{"key":"e_1_2_1_27_1","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-71-12-3027"},{"key":"e_1_2_1_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/0140-6736(90)90134-Q"},{"key":"e_1_2_1_29_1","first-page":"360","volume-title":"Viral Hepatitis and Liver Disease","author":"Weiner AJ","year":"1991"},{"key":"e_1_2_1_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(89)90066-4"}],"container-title":["Journal of Medical Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fjmv.1890380312","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmv.1890380312","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,24]],"date-time":"2023-10-24T02:04:10Z","timestamp":1698113050000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jmv.1890380312"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1992,11]]},"references-count":29,"journal-issue":{"issue":"3","published-print":{"date-parts":[[1992,11]]}},"alternative-id":["10.1002\/jmv.1890380312"],"URL":"https:\/\/doi.org\/10.1002\/jmv.1890380312","archive":["Portico"],"relation":{},"ISSN":["0146-6615","1096-9071"],"issn-type":[{"value":"0146-6615","type":"print"},{"value":"1096-9071","type":"electronic"}],"subject":[],"published":{"date-parts":[[1992,11]]}}}