| Sustained high-throughput polymerase chain reaction diagnostics during the European epidemic of Bluetongue virus serotype 8. | |
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MedLine Citation:
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PMID: 22529113 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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A real-time reverse transcription polymerase chain reaction assay (PCR test) based on genome segment 10 of Bluetongue virus (BTV) was developed. The PCR test consists of robotized viral RNA isolation from blood samples and an all-in-one method including initial denaturation of genomic double-stranded RNA, reverse transcription polymerase chain reaction (RT-PCR), and real-time detection and analysis. Reference strains of the 24 recognized BTV serotypes, isolates from different years, and geographic origins were detected. Other orbiviruses such as African horse sickness virus, Epizootic hemorrhagic disease virus, and Equine encephalosis virus were not detected. Experimentally infected animals were PCR positive from 2 days postinoculation, which was earlier than fever, other clinical signs, or seroconversion. The diagnostic sensitivity and specificity were very close to or even 100%. The PCR test played a key role in the detection of BTV serotype 8 in August 2006 in The Netherlands. The outbreak in a completely naive ruminant population allowed for further evaluation of the PCR test with field samples. In 2006, the correlation between enzyme-linked immunosorbent assay and PCR results was estimated to be 95%. In the following years, the PCR test was used for diagnosis of diseased animals, for testing of healthy animals for trade purposes, and for detection of BTV RNA in different species of the insect vector, Culicoides. In the autumn of 2008, BTV serotype 6 unexpectedly emerged in northwest Europe and was also detected with the PCR test developed in the current study. The performance in routine use over 5 years has been recorded and evaluated. |
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Authors:
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Piet A van Rijn; René G Heutink; Jan Boonstra; Hans A Kramps; René G P van Gennip |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc Volume: 24 ISSN: 1943-4936 ISO Abbreviation: J. Vet. Diagn. Invest. Publication Date: 2012 May |
Date Detail:
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Created Date: 2012-04-24 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9011490 Medline TA: J Vet Diagn Invest Country: United States |
Other Details:
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Languages: eng Pagination: 469-78 Citation Subset: IM |
Affiliation:
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1Piet A. van Rijn, Central Veterinary Institute of Wageningen UR (CVI), PO Box 65, 8200 AB Lelystad, The Netherlands. piet.vanrijn@wur.nl. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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