鸡组织滴虫病PCR病原学诊断方法的建立  被引量:7

Development of PCR assay for detection of Histomonas meleagridis infection in chickens

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作  者:曲昌宝 王尚尚 侯照峰 宿世杰 刘丹丹 曹李琴 陶建平 许金俊 

机构地区:[1]扬州大学禽类预防医学教育部重点实验室,江苏扬州225009

出  处:《中国预防兽医学报》2013年第2期160-163,共4页Chinese Journal of Preventive Veterinary Medicine

基  金:教育部创新团队计划(IRT0978);江苏省动物预防医学省级重点实验室开放课题(K100014)

摘  要:为建立鸡组织滴虫(H.meleagridis)的PCR检测方法,本研究以确诊的感染火鸡组织滴虫的阳性鸡的肝组织DNA为模板,设计针对18S rRNA基因高度保守的特异性引物,建立组织滴虫病的PCR诊断方法。敏感性试验显示阳性样品组织DNA的最低检出限为4 ng/μL。特异性试验表明与其他常见鸡寄生虫的DNA样品均无交叉反应。该PCR方法对临床样品的检出率为100%,与组织病理学诊断结果符合率为100%。该诊断方法敏感、特异,可以用于组织滴虫病的临床诊断。To establish a rapid protocol for detecting Histomonas meleagridis infection in chickens A PCR assay was developed with a pair of specific primers designed according to 18S rRNA gene of H.meleagridis. Under the optimized reaction conditions, the specific DNA fragment (550 bp) was amplified and verified by sequence analysis from the clinical liver samples. Results of sensitivity test indicated that the detection limit of PCR was 4 ng/IxL DNA in the infected liver. The specificity test resluts showed no cross reaction with orther common parasites in chickens. Tested on the 14 suspicious clinical samples of chick livers collected from Jiangsu province, 100% were compared with histopathological diagnosis results. In routine diagnosis of H.meleagridis in poultry positive detected by the PCR assay, and the coincidence rate was 100% conclusion, the PCR assay is specific and sensitive, and could be useful for

关 键 词:火鸡组织滴虫 18S核糖体RNA基因 PCR 诊断 

分 类 号:S852.72[农业科学—基础兽医学]

 

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