敏感性PCR检测猪肉弓形虫方法的建立与应用  

Establishment and application on a sensitive PCR assay for detection of Toxoplasma gondii in retail pork

作　　者：余莉[1] 朱新伊[1] 胡元生 张俊玲[1] 罗庆礼[1] 沈继龙[1] 

机构地区：[1]安徽医科大学病原生物学教研室,人兽共患病安徽省重点实验室,合肥230032 [2]安徽医科大学第三附属医院检验科,合肥230031

出　　处：《中国人兽共患病学报》2010年第10期895-899,共5页Chinese Journal of Zoonoses

基　　金：国家自然科学基金青年基金(No.30800965);安徽省教育厅高校自然科学基金(No.KJ2008B291)联合资助

摘　　要：目的建立一种敏感快速的猪肉弓形虫PCR检测方法,并用来检测市售猪肉弓形虫的感染情况。方法采用不同数量的弓形虫模拟感染猪肉样本,分别以529 bp重复序列、B1基因、SAG1、SAG2、SAG3作为目标检测基因,对其敏感性进行评价,在此基础上运用敏感的靶标检测100份市售猪肉样本中弓形虫的感染情况。结果 529 bp重复序列的敏感性最高,用此靶标检测合肥市售猪肉,弓形虫阳性率为17%。结论 529 bp重复序列基因是一个可用于肉制品中弓形虫PCR检测的理想靶标。A sensitive PCR assay for detection of Toxoplasma gondii（T.gondii） was developed and evaluated in retail pork infeeted with T.gondii.Pork samples were contaminated artificially by different amounts of T.gondii tachyzoite（0-10 000）,and the 529 bp repetitive sequence（Toxo-529）,B1 gene,SAG1,SAG2,and SAG3 of T.gondii were used as detecting targets respectively.Sensitivity of target genes was evaluated,and results indicated that the 529 bp repetitive sequence was the most sensitive target among these candidate genes.Then,100 retail pork samples were collected to analyze T.gondii contamination and the positive rate for T.gondii DNA fragments was about 17%.These results demonstrate that the 529 bp repetitive sequence is a sensitive target gene for PCR detection of T.gondii in retail pork.

关 键 词：弓形虫 PCR 猪肉 

分 类 号：R382[医药卫生—医学寄生虫学]