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海洋通报  2012 

采用ICC-qPCR法分析渤海湾表层海水中的轮状病毒

DOI: 10.11840/j.issn.1001-6392.2012.3.011, PP. 308-313

Keywords: 轮状病毒,感染性,ICC-qPCR,渤海湾,表层海水

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Abstract:

轮状病毒是引起婴幼儿急性腹泻的重要病原体之一,在水环境中存活时间长,导致人类感染的剂量低,因此寻求一种快速高效的定量检测海水中的轮状病毒方法势在必行。传统的细胞培养技术不但耗时,而且灵敏度低,现代分子生物学技术虽然克服了上述缺点,但是其感染性的信息无从获得。因此,本文建立了细胞培养结合实时定量PCR(ICC-qPCR)的方法,并于2010年冬季对渤海湾天津近岸重点海域表层海水中具有感染性的轮状病毒进行了定量调查。500mL海水经浓缩,48h细胞培养之后,用qPCR方法从7个海水样品中检测出3个样品具有感染性,其测定值范围为1.8×102copies~3.8×103copies,该海域感染性轮状病毒的含量为1~39PFU/L。ICC-qPCR方法快速、灵敏,将细胞培养与实时定量PCR技术相结合,能对感染性病毒进行定量分析,因此有望在今后的水环境胃肠道病毒的监测中得到广泛应用。

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