| Detection of norovirus capsid proteins in faecal and food samples by a real time immuno-PCR method. | |
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MedLine Citation:
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PMID: 16478496 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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AIMS: To develop a sensitive real time immuno-polymerase chain reaction (rtI-PCR) method for detecting norovirus (NV) capsid protein in food samples. METHODS AND RESULTS: The viral antigens were captured by two polyclonal antisera against recombinant Norwalk viral-like particles (rNVLPs). Biotin-conjugated antibodies, avidin and biotin-conjugated DNA reporter were used to convert the protein signals into DNA signals. The reporter DNA was then amplified by addition of primers and PCR. A real time PCR method was used in order to perform a quantitative post-PCR analysis. One hundred rNVLPs (10 fg) and a NV sample containing 660 rNVLPs equivalent particle units (66 fg) could be detected by this method. CONCLUSION: The PCR inhibitors present in the food samples had minimal effect on antigen capture and were removed by multiple wash steps during the rtI-PCR procedure. The sensitivity of rtI-PCR was >1000-fold higher than the standard enzyme-linked immunosorbent assay and approximately 10 times higher than reverse transcription PCR in detection of NV capsid protein in stool and food samples. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of a rtI-PCR method to detect NV in contaminated food samples without concentration or purification of the virus. |
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Authors:
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P Tian; R Mandrell |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of applied microbiology Volume: 100 ISSN: 1364-5072 ISO Abbreviation: J. Appl. Microbiol. Publication Date: 2006 Mar |
Date Detail:
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Created Date: 2006-02-15 Completed Date: 2006-08-23 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9706280 Medline TA: J Appl Microbiol Country: England |
Other Details:
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Languages: eng Pagination: 564-74 Citation Subset: IM |
Affiliation:
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United States Department of Agriculture, Agricultural Research Service, Produce Safety and Microbiology Research Unit, Western Regional Research Center, Albany, CA 94710-1105, USA. ptian@pw.usda.gov |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Antigens, Viral / analysis Capsid Proteins / analysis*, immunology Enzyme-Linked Immunosorbent Assay / methods Feces / chemistry*, virology Food Analysis / methods* Food Contamination / analysis Food Microbiology Fragaria / chemistry, immunology Norovirus / chemistry*, immunology Ostreidae / chemistry, immunology Polymerase Chain Reaction / methods RNA, Viral / analysis Recombinant Proteins / analysis, immunology Reverse Transcriptase Polymerase Chain Reaction / methods Shellfish / analysis |
| Chemical | |
Reg. No./Substance:
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0/Antigens, Viral; 0/Capsid Proteins; 0/Norwalk virus capsid; 0/RNA, Viral; 0/Recombinant Proteins |
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