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曾爱华, 梅寒芳, 杨小蓉, 金小宝, 朱家勇. 贝类GⅠ型扎幌样病毒荧光定量PCR检测[J]. 中国公共卫生, 2009, 25(10): 1227-1228. DOI: 10.11847/zgggws2009-25-10-39
引用本文: 曾爱华, 梅寒芳, 杨小蓉, 金小宝, 朱家勇. 贝类GⅠ型扎幌样病毒荧光定量PCR检测[J]. 中国公共卫生, 2009, 25(10): 1227-1228. DOI: 10.11847/zgggws2009-25-10-39
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ZENG Ai-hua, MEI Han-fang, YANG Xiao-rong, . Development of fluorescence quantitative PCR for detection of genogroup Ⅰ Sapporo-like viruses in shellfish[J]. Chinese Journal of Public Health, 2009, 25(10): 1227-1228. DOI: 10.11847/zgggws2009-25-10-39
Citation: ZENG Ai-hua, MEI Han-fang, YANG Xiao-rong, . Development of fluorescence quantitative PCR for detection of genogroup Ⅰ Sapporo-like viruses in shellfish[J]. Chinese Journal of Public Health, 2009, 25(10): 1227-1228. DOI: 10.11847/zgggws2009-25-10-39
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贝类GⅠ型扎幌样病毒荧光定量PCR检测

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

    摘要
  • 摘要: 目的建立贝类中GⅠ型扎幌样病毒的荧光定量PCR检测方法。方法通过用聚乙二醇6000(PEG6000)对贝类中扎幌样病毒进行浓缩,用序列比对软件设计出针对GⅠ型扎幌样病毒保守序列的通用引物与探针,建立GⅠ型扎幌样病毒荧光定量PCR检测方法。结果浓度为106~104,103,102~101拷贝/μL的GⅠ扎幌样病毒检出率分别为3/3,1/3,0/3,因此,常规逆转聚合酶链反应(RT-PCR)最低检出限为103拷贝/μL。此荧光定量PCR方法对贝类中GⅠ型扎幌样病毒检测高度特异,最低检出限可达102拷贝/μL,比常规RT-PCR的低1个数量级,即灵敏度高10倍,线性范围为102~106拷贝/μL,标准曲线的相关系数为0.998 8。结论本研究建立了GⅠ型扎幌样病毒的荧光定量PCR检测方法,可用于贝类中GⅠ型扎幌样病毒污染状况及其突发事件的快速定量检测。

     

    Abstract: 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.

     

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