摘要
兔球虫病由艾美耳属球虫寄生于消化道而引起，对养兔业的危害极大。由于兔艾美耳球虫的11个虫种在致病性上有较大差异，因此进行准确的虫种鉴定对兔球虫病的诊断和防治有十分重要的意义。为提高和扩大PCR方法检测粪便样品中艾美耳球虫的灵敏度和应用范围，我们利用全基因组扩增技术对提取自粪便样品的球虫基因组进行预扩增，提高PCR反应初始模板含量，再利用兔艾美耳球虫ITS-1序列种特异性引物对WGA产物进行PCR扩增，以鉴定虫种。结果显示，经全基因组扩增后的样品的PCR检测能够在单个卵囊的水平上进行虫种鉴定。利用该方法，我们对10份卵囊含量较少的田间兔粪样品进行了PCR检测，对其中的虫种进行了的鉴定。检测结果显示，与卵囊形态鉴定相比，该方法不仅与能确定形态的形态学检测结果一致，还能够检测出形态学不易判断的虫种，体现了更高的准确性。该技术的应用可提高田间样品检测的敏感性和准确度，为兔球虫临床感染情况提供实践指导。
Eimeria spp. of rabbits which cause great economic losses in rabbit industry belong to Phylum Apicomplexa. Since rabbit coccidia contain 11 species, which have different pathogenicity between each other, it is necessary to identify the exact species in the feild samples to control rabbit coccidiosis. In order to improve the sensitivity and expand the application of PCR detection, we introduced the Whole Genome Amplification technique to preamplify the DNA template extracted from oocysts, then WGA product were used to amplify the ITS-1 sequence to identify species. The results showed that the optimized method guaranteed higher sensitivity which can identify the species based on single oocyst. Detection of 10 filed samples showed excellent specificity and consistency with morphological identification. By this method, the species that hard to discriminate under microscope can be accurately identified. Application of this method which provides higher sensitivity and specificity could make great effort to the rabbit coccidiosis investigation in practice.
出处
《寄生虫与医学昆虫学报》
CAS
2014年第4期268-272,共5页
Acta Parasitologica et Medica Entomologica Sinica
基金
国家兔产业技术体系专项基金（CARS44）
