Leng X, Mosier D A, Oberst R D
Department of Diagnostic Medicine-Pathology, College of Veterinary Medicine, Kansas State University, Manhattan 66506-5606, USA.
Appl Environ Microbiol. 1996 Feb;62(2):643-7. doi: 10.1128/aem.62.2.643-647.1996.
An assay to identify Cryptosporidium DNA in bovine feces has been developed emphasizing standardization of sample preparation and simplification of the DNA recovery process for PCR amplification and DNA hybridization detection. The Cryptosporidium DNA recovery-PCR detection procedure (CR-PCR) can recover DNA suitable for PCR amplification without using or generating hazardous chemicals or wastes. In comparisons with a commercial enzyme-linked immunoassay (Color Vue-Cryptosporidium; Seradyn, Indianapolis, Ind.), the CR-PCR could detect 10(3) to 10(4) times fewer purified oocysts diluted in solution (water or buffered saline) and 10(2) times fewer oocysts from diarrheic feces and showed earlier detectability from solid, nondiarrheic feces in an experimental infection. This assay may prove useful for detecting Cryptosporidium oocysts in feces and in clarifying the role of livestock in waterborne outbreaks of cryptosporidiosis.
已开发出一种用于鉴定牛粪便中隐孢子虫DNA的检测方法，该方法着重于样品制备的标准化以及用于PCR扩增和DNA杂交检测的DNA回收过程的简化。隐孢子虫DNA回收-PCR检测程序（CR-PCR）无需使用或产生有害化学物质或废物，就能回收适合PCR扩增的DNA。与商用酶联免疫分析（Color Vue-隐孢子虫；Seradyn公司，印第安纳波利斯，印第安纳州）相比，CR-PCR能够检测出溶液（水或缓冲盐水）中稀释的纯化卵囊数量少10³至10⁴倍、腹泻粪便中卵囊数量少10²倍，并且在实验性感染中能更早地从固体、非腹泻粪便中检测到。该检测方法可能有助于检测粪便中的隐孢子虫卵囊，并阐明家畜在水源性隐孢子虫病暴发中的作用。
