多重PCR检测3种鸡球虫方法的建立  

Establishment of a multiplex PCR assay to detect Eimerim maxima,E. tenella and E. acervulina

作　　者：辛玲[1] 许金俊[1] 陶建平[1] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009

出　　处：《中国兽医杂志》2008年第3期6-8,共3页Chinese Journal of Veterinary Medicine

基　　金：江苏省教育厅课题(NK0310078);江苏省高校重点实验室开放课题(KJS01056)

摘　　要：根据GenBank中发表的巨型艾美耳球虫、柔嫩艾美耳球虫和堆型艾美耳球虫TS-1序列,设计了3对引物,建立了这3种球虫的单一PCR和多重PCR检测方法,并分别对单一PCR和多重PCR方法的特异性和敏感性进行了研究,对巨型艾美耳球虫、柔嫩艾美耳球虫和堆型艾美耳球虫的混合卵囊进行了初步应用。结果显示:单一PCR和多重PCR均能扩增出巨型艾美耳球虫、柔嫩艾美耳球虫和堆型艾美耳球虫特异性条带,其大小分别为151 bp、463 bp、303 bp,其最小检测浓度为0.5ng,对水牛梭形肉孢子虫、有毒艾美耳球虫、猪源弓形虫汤山株及其田间分离株均不起反应,表明建立的方法具有很强的特异性和较高的敏感性,可望用于巨型艾美耳球虫、柔嫩艾美耳球虫和堆型艾美耳球虫的诊断和田间种类调查。Three sets of spectific primers were designed according to the previously published ITS-1 sequences of Eimeria maxima, E. tenella and E. acervulina in the GenBank. A single PCR and multiplex PCR methods were established, and used to detect the mixed samples which contained E. maxima, E. tenella and E. acervulina. The results showed that both methods amplified the special fragments with 151 bp, 463 bp and 303 bp from E. maxima, E. tenella and E. acervulina, respectively, and there were no cross reactions with Sarcocystis fusiformis, Toxoplasma gondii and E. noces. The lowest concentration of DNA that could be detected was 0.5 ng. These results indicated that the multiplex PCR was a rapid, sensitive and specific method for detecting Eimeria spp. in the field.

关 键 词：鸡球虫 多重PCR检测 

分 类 号：S855.9[农业科学—临床兽医学]