聚合酶链反应检测隐孢子虫的临床应用探讨

目的　探讨一种更为简便的聚合酶链反应（PCR）检测人体微小隐孢子虫（Cryptosporidiumparvum,C.p.）的模板制备方法。方法　根据LaxerMA等克隆的C.p.核酸序列片段设计并合成一对引物，将含有C.p.卵囊的粪便经浓集、裂解后，直接取上清液作为模板与引物进行PCR反应。　结果　取自于桂林、福州和徐州三地含有C.p.的粪便标本均能扩增出418bp的C.p.目的DNA条带，阴性对照无扩增。结论　应用该引物对人粪便中的C.p.进行PCR检测具有高度特异性,该模板制备方法能缩短PCR检测C.p.时间，较适于临床实际应用。

EXPLORATION ON THECLINICAL DETECTION OF CRYPTOSPORIDIUM PARVUM BY THE POLYMERASE CHAIN REACTION

Objective To explore a more simple and convenient DNA-template-maken method detecting Cryptosporidium parvum with the polymerase chain reaction (PCR)　Methods A pair of primers were designed and synthesized according to the nucleotide sequence fragment of C. parvum cloned by Laxer MA. DNA template obtained directly from childrens fecal samples supernatant with C. parvum oocysts which were concentrated and split, then put the templates and the primers to PCR.　Results　The fecal samples with C. parvum oocysts obtained from Guilin, Fuzhou and Xuzhou respectively could been amplified out 418 bp target DNA fragments of the C. parvum. Negative control showed no amplification.　Conclusion　PCR detected C. parvum of human feces with the pair of primers showed higher sensitivity, the template-make-method could shorten the time which PCR detected C. parvum, and more suited to clinical practice.