Leng X, Mosier D A, Oberst R D
Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan 66506-5605, USA.
Vet Parasitol. 1996 Mar;62(1-2):1-7. doi: 10.1016/0304-4017(95)00863-2.
We have developed a combined polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assay for the detection and differentiation of Cryptosporidium parvum, Cryptosporidium muris, and Cryptosporidium baileyi. The assay utilizes PCR to amplify an approximately 1750 basepair product of the Cryptosporidium 18S rRNA gene. Following double digestion with restriction endonucleases Dral and Vsp1, this PCR product yields DNA fragments that can be resolved electrophoretically into RFLP profiles that are distinct for C. parvum, C. muris, and C. baileyi. Previous PCR-restriction analyses could not simultaneously differentiate all three species. Future application of this technique could include predicting the disease-causing potential of oocyst-contaminated environmental specimens and helping to determine the source of oocyst contamination.
我们开发了一种聚合酶链反应（PCR）-限制性片段长度多态性（RFLP）联合检测方法，用于检测和区分微小隐孢子虫、鼠隐孢子虫和贝氏隐孢子虫。该检测方法利用PCR扩增隐孢子虫18S rRNA基因约1750个碱基对的产物。用限制性内切酶Dral和Vsp1进行双酶切后，该PCR产物产生的DNA片段可通过电泳分离成RFLP图谱，这些图谱对于微小隐孢子虫、鼠隐孢子虫和贝氏隐孢子虫是不同的。以前的PCR-限制性分析不能同时区分这三种物种。该技术未来的应用可能包括预测受卵囊污染的环境样本的致病潜力，并有助于确定卵囊污染的来源。
