| Comparison of methods for improved RNA extraction from blood for early detection of Classical swine fever virus by real-time reverse transcription polymerase chain reaction. | |
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MedLine Citation:
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PMID: 21908315 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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Classical swine fever (CSF) is a highly contagious disease of pigs. Early detection of the Classical swine fever virus (CSFV) in infected animals and routine surveillance is important for a rapid response and control of an outbreak of CSF. The current study investigated whole blood as a clinical specimen for the detection of CSFV by real-time reverse transcription polymerase chain reaction (real-time RT-PCR) in experimentally infected pigs. The virus was detectable in pre-clinical animals in whole blood and in different fractions of blood, including white blood cells, red blood cells (RBC), and serum. Based on an in-vitro binding assay, CSFV is retained in the RBC fraction. Naturally occurring PCR inhibitors of whole blood were shown to inhibit detection, and several commercial RNA extraction kits failed to remove these inhibitors. The commercial blood RNA extraction protocols were modified to achieve optimized single tube and high-throughput 96-well plate RNA extraction that efficiently removed PCR inhibitors from whole blood and enhanced detection of CSFV in experimentally inoculated pigs. This enabled detection 1-2 days earlier than observed using unmodified RNA extraction protocols. The results show effective use of whole blood as a clinical specimen for diagnosis and surveillance of CSF in pre-clinical animals. |
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Authors:
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Amaresh Das; Tammy R Beckham; Michael T McIntosh |
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Publication Detail:
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Type: Journal Article Date: 2011-06-08 |
Journal Detail:
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Title: Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc Volume: 23 ISSN: 1943-4936 ISO Abbreviation: J. Vet. Diagn. Invest. Publication Date: 2011 Jul |
Date Detail:
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Created Date: 2011-09-12 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: 727-35 Citation Subset: IM |
Affiliation:
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1Michael T. McIntosh, PO Box 848, Plum Island Animal Disease Center, Foreign Animal Disease Diagnostic Laboratory, USDA, APHIS, VS, NVSL, Greenport, NY 11944-0848. michael.t.mcintosh@aphis.usda.gov. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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