摘要:目的 了解生蔬食品及相关水源微小隐孢子虫污染情况，并研究其卵囊分离及18S rRNA鉴定的方法。 方法 采用分步离心富集法从78份生蔬食品和相关水源中分离出微小隐孢子虫卵囊并提取DNA模板，利用微小隐孢子虫18S rRNA的基因序列设计特异性引物(446bp)进行多聚酶链反应扩增，扩增产物经回收克隆后进行PCR鉴定、酶切鉴定及序列同源性分析。 结果 从78份样品中检出4份特异性条带样品。限性样品的重组克隆经PCR鉴定可重现446bp的特异性条带，其酶切产物亦与目的基因PCR产物位置相同。重组菌液的测序结果与Geabank数据库中微小隐孢子虫18S rRNA基因序列的同源性为99%。 结论 建立了一种监测生蔬食品及相关水源中微小隐孢子虫污染的基因检测方法。

    关键词:微小隐孢子虫;18S rRNA;聚合酶链反应

    Identification of Cryptosporidium parvum18S rRNA in food and water.

    GENG Yi-jie，GAO Shi-tong，HUANG Da-na，et al.

    (Shenzhen Municipal Center for Disease Control and Prevention，Shenzhen518020，Guangdong，P.R.China)

    Abstract:Objective To isolate Cryptosporidium parvumoocysts from food and water and identify its18S rRNA. Methods 78food or water samples were enriched and Cryptosporidium parvumOocysts was isolated.DNA was extracted and specific primers for the18S rRNA of Cryptosporidium parvumwas used in polymerase chain reaction(PCR).The amplified products were cloned and analyzed by PCR，digested with restriction enzyme and sequenced. Results 446bp PCR products were observed by agarose gel electrophoresis from the78samples.After cloning ......