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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
2
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pubmed:dateCreated |
1995-5-18
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pubmed:abstractText |
We describe a simple method for the rapid detection of bovine viral diarrhea virus (BVDV) that uses a one-tube reverse transcription PCR (RT-PCR) and total RNA extracted directly from a variety of bovine specimens, including whole blood and tissues. Reagents for both RT and PCR were combined in a one-tube, single-buffer system, and amplification was performed with a single uninterrupted thermal cycling program. Using the novel cationic surfactant tetradecyltrimethylammonium oxalate (Catrimox-14), we consistently extracted RT-PCR-quality RNA from specimens containing blood. Amplification with primers derived from conserved sequences within the BVDV 5'-untranslated region yielded a 244-bp product. Assay specificity was confirmed by ethidium bromide-stained gel electrophoresis and by chemiluminescence-assayed Southern blot hybridizations involving BVDV 5'-untranslated region-specific digoxigenin-labelled cDNA probes. The assay detection level was 0.1 50% tissue culture infectious dose of BVDV when ethidium bromide-stained gel electrophoresis was used and 0.01 50% tissue culture infectious dose of BVDV when Southern blot hybridization was used. Our method is an alternative to the conventional cell culture assays used in a diagnostic laboratory and is an improvement over existing RT-PCR assays for BVDV.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1318605,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1325330,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1329326,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1333126,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1361933,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1381195,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1503745,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1645593,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1655059,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1656820,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1665207,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-167506,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1709950,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1715173,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-1885726,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-2156132,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-2175527,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-2419063,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-2440339,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-2562207,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-271968,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-2838957,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-2844971,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-6184674,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-7678747,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-7680772,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-7685192,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-7689852,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-8011781,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-8136141,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-8290362,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-8292344,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-8384625,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-8385157,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7714180-8431290
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Feb
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pubmed:issn |
0095-1137
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
33
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
287-91
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:7714180-Animals,
pubmed-meshheading:7714180-Base Sequence,
pubmed-meshheading:7714180-Bovine Virus Diarrhea-Mucosal Disease,
pubmed-meshheading:7714180-Cattle,
pubmed-meshheading:7714180-DNA, Viral,
pubmed-meshheading:7714180-DNA Primers,
pubmed-meshheading:7714180-Detergents,
pubmed-meshheading:7714180-Diarrhea Viruses, Bovine Viral,
pubmed-meshheading:7714180-Evaluation Studies as Topic,
pubmed-meshheading:7714180-Molecular Sequence Data,
pubmed-meshheading:7714180-Polymerase Chain Reaction,
pubmed-meshheading:7714180-Quaternary Ammonium Compounds,
pubmed-meshheading:7714180-RNA, Viral,
pubmed-meshheading:7714180-Sensitivity and Specificity,
pubmed-meshheading:7714180-Transcription, Genetic,
pubmed-meshheading:7714180-Trimethyl Ammonium Compounds,
pubmed-meshheading:7714180-Virology
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pubmed:year |
1995
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pubmed:articleTitle |
Rapid detection of bovine viral diarrhea virus by using RNA extracted directly from assorted specimens and a one-tube reverse transcription PCR assay.
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pubmed:affiliation |
Virology Laboratory, Manitoba Agriculture Veterinary Services, Winnipeg, Canada.
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pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, Non-U.S. Gov't
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