Bonnin A, Fourmaux M N, Dubremetz J F, Nelson R G, Gobet P, Harly G, Buisson M, Puygauthier-Toubas D, Gabriel-Pospisil G, Naciri M, Camerlynck P
Laboratoire de Parasitologie et Mycologie, Centre Hospitalier Universitaire de Dijon, France.
FEMS Microbiol Lett. 1996 Apr 1;137(2-3):207-11. doi: 10.1111/j.1574-6968.1996.tb08107.x.
In order to define transmission routes of cryptosporidiosis and develop markers that distinguish Cryptosporidium parvum isolates, we have identified 2 polymorphic restriction enzyme sites in a C. parvum repetitive DNA sequence. The target sequence was amplified by polymerase chain reaction from 100 to 500 oocysts and the amplified product was subjected to restriction enzyme digestion. Typing of 23 isolates showed that 10/10 calf isolates had the same profile. In contrast, 2 patterns were observed among human isolates: 7/13 displayed the calf profile, and 6/13 presented another pattern. The PCR-RFLP assay described here is a sensitive tool to distinguish C. parvum isolates.
为了确定隐孢子虫病的传播途径并开发区分微小隐孢子虫分离株的标志物，我们在微小隐孢子虫重复DNA序列中鉴定出2个多态性限制性酶切位点。通过聚合酶链反应从100至500个卵囊中扩增目标序列，并对扩增产物进行限制性酶切消化。对23个分离株进行分型显示，10株犊牛分离株具有相同的图谱。相比之下，在人类分离株中观察到2种模式：13株中有7株显示出犊牛图谱，13株中有6株呈现另一种模式。本文所述的PCR-RFLP分析是区分微小隐孢子虫分离株的灵敏工具。
