Source:http://linkedlifedata.com/resource/pubmed/id/11011076
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
2000-11-15
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pubmed:abstractText |
Epidemiological evidence linking the transmission of enteric viral disease to shellfish has been known for a long time. A variety of methods have been described for the detection of viral contaminants in shellfish using RT-PCR. However, these methods generally include numerous, often fastidious and time consuming steps for virus release from shellfish tissues and viral RNA isolation. A simplified procedure based on the enzymatic liquefaction of shellfish digestive tissues without any mechanical homogenisation step, followed by a simple clarification of the lysate using dichloromethane extraction, was developed. Viral RNA is isolated directly from the shellfish extract by a guanidium thiocyanate-silica extraction method, adapted for the use of a vacuum manifold system. Virus-specific RT-PCR assays were set up for detection of genomic sequences of the predominant viral pathogens, HAV, Astrovirus and Norwalk-like viruses (from genogoups I or II). The specificity of the amplicons is confirmed finally by hybridisation with DIG-labelled specific probes. The overall procedure applied to shellfish samples spiked with HAV particles allowed a detection of 20 pfu of HAV per g of hepatopancreas. In addition, up to 20 samples can be tested within 24 h.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
0166-0934
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
90
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1-14
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:11011076-Animals,
pubmed-meshheading:11011076-Bivalvia,
pubmed-meshheading:11011076-Hepatovirus,
pubmed-meshheading:11011076-Immunoblotting,
pubmed-meshheading:11011076-Mamastrovirus,
pubmed-meshheading:11011076-Norwalk virus,
pubmed-meshheading:11011076-Ostreidae,
pubmed-meshheading:11011076-RNA, Viral,
pubmed-meshheading:11011076-RNA Viruses,
pubmed-meshheading:11011076-Reverse Transcriptase Polymerase Chain Reaction,
pubmed-meshheading:11011076-Sensitivity and Specificity,
pubmed-meshheading:11011076-Shellfish
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pubmed:year |
2000
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pubmed:articleTitle |
Simplified procedure for detection of enteric pathogenic viruses in shellfish by RT-PCR.
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pubmed:affiliation |
Service R&D, Institut Pasteur de Lille, 1 rue du Professeur Calmette, BP 245, 59019 Cedex, Lille, France.
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pubmed:publicationType |
Journal Article
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