Development of fluorescence quantitative PCR for detection of genogroup Ⅰ Sapporo-like viruses in shellfish

ObjectiveTo develop a fluorescence quantitative polymerase chain reaction(FQ-PCR)for the detection of geno group I(GI)Sapporo-like viruses(SLVs)in shellfish.MethodsSLVs were concentrated by PEG 6000.The degenerate primers and probe were designed following large scale SLVs genome consensus analysis and subsequently a FQ-PCR assay for detection of GISLVs was established.ResultsT the assay developed possessed high accuracy and repetition for SLVs detection.The sensitivity of the assay was as low as 10² copies per reaction.The assay was linear within 5-log dynamic range between 10² copies and 10⁶ copies.The correlation coefficient of the standard curve was 0.9988.ConclusionThe detection method of geno group I Sapporo-like viruses was established with FQ-PCR and can be used for rapid detection of SLVs pollution in shellfish and emergency of SLVs.