犬源环孢子虫PCR检测方法的建立  

Development of a specific PCR assay for detecting Cyclospora sp. in dogs

作　　者：岳彩玲[1] 李国清[1] 程家林[1] 高振永[1] 刘霞[1] 朱海波[1] 张萍[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽医科学》2010年第11期1128-1131,共4页Chinese Veterinary Science

基　　金：国家自然科学基金项目(30671577)

摘　　要：为了建立犬源环孢子虫的PCR检测方法,根据该环孢子虫18SrDNA基因序列,设计1对特异性引物;通过阳性参照摸索出该检测方法的最佳反应条件,进行了特异性和敏感性试验;并对临床样品进行了检测。结果显示,该PCR检测方法能特异性地扩增出犬源环孢子虫18SrDNA中379bp的片段,与牛源环孢子虫、柔嫩艾美耳球虫、人隐孢子虫、蓝氏贾第虫、刚第弓形虫、毛首线虫、大片吸虫、犬弓首蛔虫等8种虫体基因组DNA无交叉反应;最低能检测到84fg的阳性参照DNA,检测的157份临床样品中8份为阳性,比传统镜检方法多出3份。结果表明,建立的PCR检测方法具有较高的特异性和敏感性,可用于犬环孢子虫的临床检测。To establish a specific PCR assay for detecting Cyclospora sp. in dogs,a pair of primers was designed to amplify a 379bp DNA fragment according to the 18S rDNA sequence of the dog-derived Cyclospora sp..With positive template,PCR reaction condition was optimized.A series of experiments on specificity,sensitivity and clinical application were conducted.The PCR assay was specific and there was no cross-reaction with other parasites,such as cattle-derived Cyclospora sp,Eimeria tenella,Cryptosporidium hominis,Giardia lamblia,Toxoplasma gondii,Trichuris sp.,Fasciola gigantica and Toxocara canis.The assay was able to detect as low as 84fg control positive DNA.The results of clinical samples detection revealed that the positive rate of the PCR assay was higher than that of the traditional method.It was concluded that the PCR assay was specific and sensitive,and it was an effective tool for detecting Cyclospora sp. in canine feces.

关 键 词：环孢子虫 犬 特异PCR 

分 类 号：S852.723[农业科学—基础兽医学]