{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T09:25:48Z","timestamp":1784193948626,"version":"3.55.0"},"reference-count":0,"publisher":"American Society for Microbiology","issue":"3","license":[{"start":{"date-parts":[[1997,3,1]],"date-time":"1997-03-01T00:00:00Z","timestamp":857174400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[1997,3]]},"abstract":"<jats:p>A reverse transcription-nested PCR (RT-nPCR) method for prenatal diagnosis of rubella virus (RV) infection was developed. In the first step of RT-nPCR a synthetic RNA molecule (pRRV) differing from the RV target sequence by having a 21-nucleotide insertion was used as the internal control of amplification for the detection of PCR inhibitors. In addition, comparison of pRRV and RV-specific PCR signals allowed for the semiquantitation of RV input target sequences (range, 10 to &gt; and = 1,000 RV genomes). In parallel, a complete RT-nPCR assay was performed with the same samples in the absence of the internal control to confirm the results of the first step and to detect RV RNA-positive samples containing &lt; 10 RV genomes. Subsequently, the RT-nPCR method was used to examine retrospectively clinical samples (direct RT-nPCR) from eight congenitally infected and eight uninfected fetuses for RV RNA. RT-nPCR was also used to detect RV RNA in cell cultures (culture-RT-nPCR) 96 h after inoculation with the same specimens. With amniotic fluid (AF) samples, direct RT-nPCR identified eight of eight cases of RV transmission (sensitivity, 100%), whereas culture-RT-nPCR and virus isolation detected only six of eight cases (sensitivity, 75%). However, when the culture-RT-nPCR results were positive, culture-RT-nPCR confirmed the direct RT-nPCR results 3 days to 3 weeks earlier than virus isolation. The specificity of direct RT-nPCR was 100%, with eight of eight uninfected fetuses being negative. Semiquantitation showed only small amounts (&lt; and = 100 copies) of viral RNA in clinical samples. In conclusion, direct RT-nPCR with AF samples (i) shows 100% sensitivity and specificity for prenatal diagnosis of RV infection and (ii) is a rapid technique, giving results in 24 to 48 h after sampling.<\/jats:p>","DOI":"10.1128\/jcm.35.3.708-713.1997","type":"journal-article","created":{"date-parts":[[2020,1,6]],"date-time":"2020-01-06T21:14:22Z","timestamp":1578345262000},"page":"708-713","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":52,"title":["Prenatal diagnosis of rubella virus infection by direct detection and semiquantitation of viral RNA in clinical samples by reverse transcription-PCR"],"prefix":"10.1128","volume":"35","author":[{"given":"M G","family":"Revello","sequence":"first","affiliation":[{"name":"Institute of Infectious Diseases, University of Pavia, Italy."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"F","family":"Baldanti","sequence":"additional","affiliation":[{"name":"Institute of Infectious Diseases, University of Pavia, Italy."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A","family":"Sarasini","sequence":"additional","affiliation":[{"name":"Institute of Infectious Diseases, University of Pavia, Italy."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M","family":"Zavattoni","sequence":"additional","affiliation":[{"name":"Institute of Infectious Diseases, University of Pavia, Italy."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M","family":"Torsellini","sequence":"additional","affiliation":[{"name":"Institute of Infectious Diseases, University of Pavia, Italy."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G","family":"Gerna","sequence":"additional","affiliation":[{"name":"Institute of Infectious Diseases, University of Pavia, Italy."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","container-title":["Journal of Clinical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jcm.35.3.708-713.1997","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/jcm.35.3.708-713.1997","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,3,5]],"date-time":"2022-03-05T08:16:51Z","timestamp":1646468211000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/jcm.35.3.708-713.1997"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1997,3]]},"references-count":0,"journal-issue":{"issue":"3","published-print":{"date-parts":[[1997,3]]}},"alternative-id":["10.1128\/jcm.35.3.708-713.1997"],"URL":"https:\/\/doi.org\/10.1128\/jcm.35.3.708-713.1997","relation":{},"ISSN":["0095-1137","1098-660X"],"issn-type":[{"value":"0095-1137","type":"print"},{"value":"1098-660X","type":"electronic"}],"subject":[],"published":{"date-parts":[[1997,3]]},"assertion":[{"value":"1997-03-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}