应用PCR诊断隐孢子虫病  

Diagnosis of Cryptosporidiosis by PCR

作　　者：李建华[1] 张西臣[1] 欧阳红生 尹继刚[1] 杨举[1] 李德昌[1] 

机构地区：[1]解放军农牧大学动物医学系,吉林长春130062

出　　处：《中国兽医学报》1999年第5期476-478,共3页Chinese Journal of Veterinary Science

基　　金：总后卫生部青年基金

摘　　要：应用聚合酶链反应（ Ｐ Ｃ Ｒ）建立了一种诊断人及牛等哺乳动物隐孢子虫病的方法。试验采用甘油漂浮 Ｇ３ 耐酸漏斗过滤法纯化隐孢子虫卵囊，以液氮冻融法制备模板 Ｄ Ｎ Ａ，根据隐孢子虫 １８ Ｓｒ Ｒ Ｎ Ａ 序列设计 Ｐ Ｃ Ｒ 引物建立其诊断方法。该方法特异性强，可检出鼠隐孢子虫（ Ｃｒｙｐｔｏｓｐ ｏｒｉｄｉｕｍ ｍ ｕｒｉｓ）和小球隐孢子虫（ Ｃ．ｐａｒｖｕｍ ）卵囊；敏感性高，每克粪便可检出 ４００ 个卵囊。初步应用结果表明，所建立的 Ｐ Ｃ Ｒ 方法适合于人、牛等哺乳动物隐孢子虫病的临床诊断和流行病学调查。The polymerase chain reaction (PCR)was used to amplify a 586 bp product of Cryptosporidium muris or Cryptosporidium parvum employing a pair of primers which were designed and synthesized based on the published sequence of the 18S rRNA genes in C.muris and C.parvum by computer analysis. The oocysts were purified by 1∶2 glycerine floatation and G 3 funnel filtration. The sensitivity of PCR could detected 400 oocysts per gram of feces. There were no amplification to be observed in any of Giardia lamblia, Eimeria tenella, Trypanosoma evansi and Toxoplasma gondii DNA. The preliminary application indicated that PCR was suitable for the diagnosis and survey of cryptosporidiosis.

关 键 词：隐孢子虫病 PCR 诊断 

分 类 号：R531.5[医药卫生—内科学]