Study on Reverse Transcription-Polymerase Chain Reaction Assay for Detection of Norwalk-like Virus in Shellfish

Abstract

Abstract: Objective:To explore the methods of both virus enrichment and nucleic acid extraction for establishing a Norwalk-like viruses(NL Vs) detection method by RT-PCR.Method:In this study,with poliovirus as the model to optimize glycine-PEG concentration method;The effects of 4 methods of extracting RNA from NLVs in shellfish were compared;Detection of Norwalk-like viruses in commercial shellfish samples was performed,and positive samples were demonstrated by DNA sequencing.Results:This study conducted virus enrichment with pH 9.5 glycine buffer-16% polyethylene glycol(PEG) and RNA extraction with trizol-isopropanol.The recovery of viruses was 16.8% from seeded shellfish;and the detection limit was 8.1×102 RT-PCR50/5 g shellfish.NLVs in 3 of 25 samples were found by RT-PCR and validated by DNA sequencing.Conclusion:In this study,a NLVs detection method from shellfish is established,which is sensitive and available.

Key words: Norwalk-like viruses, RT-PCR, shellfish, detection

ZHANG Ying, WU Feng-Liang. Study on Reverse Transcription-Polymerase Chain Reaction Assay for Detection of Norwalk-like Virus in Shellfish[J]. FOOD SCIENCE, 2008, 29(5): 347-351.

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Study on Reverse Transcription-Polymerase Chain Reaction Assay for Detection of Norwalk-like Virus in Shellfish

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