摘要
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 (Cryptosporidium parvum,C.p.)的模板制备方法。 方法 根据 L axer MA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 。
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 childrens 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.
出处
《中国寄生虫病防治杂志》
CSCD
2001年第1期15-16,共2页
Chinese Journal of Parasitic Disease Control
基金
广西医药卫生科研课题资助项目! (No.970 6)
