{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T14:49:54Z","timestamp":1786978194031,"version":"build-2736575974"},"reference-count":28,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2002,7,2]],"date-time":"2002-07-02T00:00:00Z","timestamp":1025568000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2002,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    A two\u2010step reverse transcriptase TaqMan\u2122 duplex PCR (RT\u2010PCR) assay was developed using the ABI 7700 Sequence Detection System for the detection of enterovirus (EV) and parechovirus type 1 and 2 (PEV) RNA from samples of cerebrospinal fluid (CSF) and throat swabs. Using sequence\u2010specific fluorescent dye labeled probes and continuous \u2018real\u2010time\u2019 monitoring, PCR amplified product accumulation was measured. Based on limiting dilutions, the TaqMan\u2122 enterovirus and parechovirus RT\u2010PCR showed an increase of two orders of magnitude compared to cell culture with sensitivity of 100% (7\/7) when assessed using enterovirus cell culture positive samples (CSF, TS). The assays were specific for enterovirus and parechovirus and did not amplify a wide selection of virus and bacterial isolates. RNA was amplified from 22 enterovirus serotypes: coxsackie A7, A9, A21; coxsackie B2, B3, B4, B5; echovirus 2, 4, 6, 7, 9, 11, 13, 17, 18, 19, 30, 31; poliovirus types 1, 2, and 3, and parechovirus types 1 and 2. The assay was used to assess the incidence of enterovirus and parechovirus RNA in cell culture negative CSF and throat swab samples (\n                    <jats:italic>n<\/jats:italic>\n                    \u2009=\u2009200). An additional 33 (15.9%) enterovirus and 2 (1%) parechovirus were identified as positive by RT\u2010PCR. Also, of 100 CSF samples from suspected cases of meningococcal meningitis submitted for meningococcal PCR testing, 59 (59%) were enterovirus and 2 (2%) parechovirus 1 and 2 were positive by RT\u2010PCR. The TaqMan\u2122 duplex assay offers a more rapid and sensitive alternative to conventional cell culture for the diagnosis of enterovirus and parechovirus infection. Closed tube real\u2010time detection using the ABI Sequence Detection System obviates the need for post\u2010PCR manipulation, which reduces hands on time and eliminates the risk of contamination from amplified PCR product. J. Med. Virol. 67:555\u2013562, 2002. \u00a9 2002 Wiley\u2010Liss, Inc.\n                  <\/jats:p>","DOI":"10.1002\/jmv.10138","type":"journal-article","created":{"date-parts":[[2002,9,10]],"date-time":"2002-09-10T22:05:28Z","timestamp":1031695528000},"page":"555-562","source":"Crossref","is-referenced-by-count":94,"title":["Development and evaluation of a \u2018real\u2010time\u2019 RT\u2010PCR for the detection of enterovirus and parechovirus RNA in CSF and throat swab samples"],"prefix":"10.1002","volume":"67","author":[{"given":"Caroline E.","family":"Corless","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Malcolm","family":"Guiver","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raymond","family":"Borrow","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valerie","family":"Edwards\u2010Jones","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew J.","family":"Fox","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edward B.","family":"Kaczmarski","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenneth J.","family":"Mutton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2002,7,2]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1136\/adc.78.4.373"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1055\/s-2007-994182"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.39.4.1553-1558.2001"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.1890290418"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1096-9071(199902)57:2<193::AID-JMV18>3.0.CO;2-N"},{"key":"e_1_2_1_7_1","first-page":"343","article-title":"Rapid detection of enteroviral RNA in cerebrospinal fluid (CSF) from patients with aseptic meningitis by reverse transcription\u2010nested polymerase chain reaction","volume":"21","author":"Furione M","year":"1998","journal-title":"Microbiologica"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-79-11-2641"},{"key":"e_1_2_1_9_1","doi-asserted-by":"publisher","DOI":"10.3109\/00365549309008542"},{"key":"e_1_2_1_10_1","first-page":"1","article-title":"Evaluation of the Applied Biosystems automated TaqMan\u2122 PCR system for the detection of meningococcal DNA","volume":"1208","author":"Guiver M","year":"2000","journal-title":"FEMS Immunol Med Microbiol"},{"key":"e_1_2_1_11_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.89.18.8847"},{"key":"e_1_2_1_12_1","doi-asserted-by":"publisher","DOI":"10.3109\/00365548909167453"},{"key":"e_1_2_1_13_1","doi-asserted-by":"publisher","DOI":"10.1097\/00006454-199711000-00015"},{"key":"e_1_2_1_14_1","first-page":"88","article-title":"Quantification of enterovirus RNA in sludge samples using single tube real\u2010time RT\u2010PCR","volume":"29","author":"Monpoeho S","year":"2000","journal-title":"BioTechniques 2000"},{"key":"e_1_2_1_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1096-9071(199907)58:3<304::AID-JMV18>3.0.CO;2-U"},{"key":"e_1_2_1_16_1","doi-asserted-by":"publisher","DOI":"10.1001\/archpedi.154.8.817"},{"key":"e_1_2_1_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1096-9071(199906)58:2<178::AID-JMV13>3.0.CO;2-Q"},{"key":"e_1_2_1_18_1","first-page":"493","article-title":"A simple method of estimating fifty percent endpoints","volume":"27","author":"Reed LJ","year":"1938","journal-title":"Am J Hygiene"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1096-9071(199610)50:2<204::AID-JMV16>3.0.CO;2-L"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.28.3.438-442.1990"},{"key":"e_1_2_1_21_1","doi-asserted-by":"publisher","DOI":"10.1086\/318452"},{"key":"e_1_2_1_22_1","doi-asserted-by":"publisher","DOI":"10.1097\/00006454-199403000-00002"},{"key":"e_1_2_1_23_1","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1128\/jcm.2.3.183-185.1975","article-title":"Propagation and isolation of group A coxsackie viruses in RD cells","volume":"2","author":"Schmidt NS","year":"1975","journal-title":"J Clin Microbiol"},{"key":"e_1_2_1_24_1","doi-asserted-by":"crossref","first-page":"230","DOI":"10.2144\/98252st02","article-title":"Comparison of three RT\u2010PCR methods","volume":"25","author":"Sellner LN","year":"1998","journal-title":"BioTechniques"},{"key":"e_1_2_1_25_1","doi-asserted-by":"publisher","DOI":"10.3109\/inf.1972.4.suppl-3.01"},{"key":"e_1_2_1_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1099-1654(200001\/02)10:1<57::AID-RMV266>3.0.CO;2-H"},{"key":"e_1_2_1_27_1","doi-asserted-by":"publisher","DOI":"10.1067\/mlc.2001.112508"},{"key":"e_1_2_1_28_1","doi-asserted-by":"publisher","DOI":"10.1002\/1531-8249(20010201)49:2<249::AID-ANA46>3.0.CO;2-3"},{"key":"e_1_2_1_29_1","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1128\/jcm.34.1.199-201.1996","article-title":"Rapid and sensitive detection of enteroviruses in specimens from patients with aseptic meningitis","volume":"34","author":"Yerly S","year":"1996","journal-title":"J Clin Microbiol"}],"container-title":["Journal of Medical Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fjmv.10138","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmv.10138","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,7]],"date-time":"2023-10-07T07:34:38Z","timestamp":1696664078000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jmv.10138"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2002,7,2]]},"references-count":28,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2002,8]]}},"alternative-id":["10.1002\/jmv.10138"],"URL":"https:\/\/doi.org\/10.1002\/jmv.10138","archive":["Portico"],"relation":{},"ISSN":["0146-6615","1096-9071"],"issn-type":[{"value":"0146-6615","type":"print"},{"value":"1096-9071","type":"electronic"}],"subject":[],"published":{"date-parts":[[2002,7,2]]}}}