| Development of SYBR Green real-time RT-PCR for rapid detection, quantitation and diagnosis of unclassified bovine enteric calicivirus. | |
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MedLine Citation:
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PMID: 19442847 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Unclassified bovine enteric calicivirus (BECV) is a newly recognized bovine enteric calicivirus that differs from bovine norovirus, and which causes diarrhea in the small intestines of calves. To date, methods such as real-time reverse transcription-polymerase chain reaction (RT-PCR) have not been developed for the rapid detection, quantitation and diagnosis of BECV. Presently, a BECV-specific SYBR Green real-time RT-PCR assay was evaluated and optimized. Diarrheic specimens (n=118) collected from 2004 to 2005 were subjected to RT-PCR, nested PCR and SYBR Green real-time RT-PCR. By conventional RT-PCR and nested PCR, 9 (7.6%) and 59 (50%) samples tested positive, respectively, whereas the SYBR Green assay detected BECV in 91 (77.1%) samples. Using BECV RNA standards generated by in vitro transcription, the SYBR Green real-time RT-PCR assay sensitively detected BECV RNA to 1.1 x 10(0)copies/microl (correlation coefficiency=0.98). The detection limits of the RT-PCR and nested PCR were 1.1 x 10(5) and 1.1 x 10(2)copies/microl, respectively. These results indicate that the SYBR Green real-time RT-PCR assay is more sensitive than conventional RT-PCR and nested PCR assays, and has potential as a reliable, reproducible, specific, sensitive and rapid tool for the detection, quantitation and diagnosis of unclassified BECV. |
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Authors:
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Sang-Ik Park; Da-Hae Park; Linda J Saif; Young-Ju Jeong; Dong-Jun Shin; Young-Hyun Chun; Su-Jin Park; Hyun-Jeong Kim; Myra Hosmillo; Hyung-Jun Kwon; Mun-Il Kang; Kyoung-Oh Cho |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2009-03-14 |
Journal Detail:
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Title: Journal of virological methods Volume: 159 ISSN: 1879-0984 ISO Abbreviation: J. Virol. Methods Publication Date: 2009 Jul |
Date Detail:
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Created Date: 2009-05-15 Completed Date: 2009-06-24 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8005839 Medline TA: J Virol Methods Country: Netherlands |
Other Details:
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Languages: eng Pagination: 64-8 Citation Subset: IM |
Affiliation:
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Bio-therapy Human Resources Center, College of Veterinary Medicine, Chonnam National University, Gwangju 500-757, South Korea. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Caliciviridae / genetics, isolation & purification* Caliciviridae Infections / diagnosis, veterinary*, virology Cattle Cattle Diseases / diagnosis*, virology Feces / virology Fluorescent Dyes Organic Chemicals* RNA, Viral / isolation & purification Reproducibility of Results Reverse Transcriptase Polymerase Chain Reaction / methods* Sensitivity and Specificity |
| Chemical | |
Reg. No./Substance:
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0/Fluorescent Dyes; 0/Organic Chemicals; 0/RNA, Viral; 163795-75-3/SYBR Green I |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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