卵形巴贝斯虫荧光定量PCR方法的建立及初步应用

发布时间：2018-03-09 17:57

本文选题：卵形巴贝斯虫　切入点：荧光定量PCR　出处：《中国兽医科学》2017年05期 　论文类型：期刊论文

【摘要】：为建立一种检测感染牛的卵形巴贝斯虫的敏感、快速方法,本研究根据GenBank中登录的卵形巴贝斯虫18S rRNA保守区设计1对特异引物,通过优化反应条件,建立了检测卵形巴贝斯虫荧光定量PCR方法。结果显示,该方法可以特异地检测出卵形巴贝斯虫DNA,对瑟氏泰勒虫、牛巴贝斯虫和双芽巴贝斯虫DNA的检测均为阴性;该方法的灵敏度可达1.26 copies/μL,比普通PCR灵敏度高100倍。组内及组间重复试验的变异系数均小于3%。对60份临床牛血液DNA样本进行检测,荧光定量PCR和常规PCR的阳性检出率分别为41.67%和33.33%。结果表明,本试验建立的荧光定量PCR方法可用于对卵形巴贝斯虫定量分析和卵形巴贝斯虫病的分子流行病学调查。
[Abstract]:In order to establish a sensitive and rapid method for the detection of Babes ovale in infected cattle, a pair of specific primers were designed according to the 18s rRNA conservative region of Babes ovum registered in GenBank, and the reaction conditions were optimized. A fluorescent quantitative PCR method was established for the detection of Babes ovale. The results showed that the method could specifically detect the DNA of Babes ovum, but the detection of DNA of Taylor serovar, Babes and Babes was negative. The sensitivity of this method can reach 1.26 copies/ 渭 L, which is 100 times higher than that of ordinary PCR. The coefficient of variation of repeated tests within and between groups is less than 3. 60 clinical bovine blood DNA samples were detected. The positive rates of fluorescent quantitative PCR and routine PCR were 41.67% and 33.33, respectively. The results showed that the fluorescent quantitative PCR method established in this study could be used for quantitative analysis of Babes ovale and molecular epidemiological investigation of oval Babes's disease.
【作者单位】：吉林农业科技学院动物科技学院;预防兽医学吉林省重点实验室;延边大学农学院;
【基金】：吉林省科技发展计划项目(20150623004TC) 吉林省重点学科培育项目(吉农院合字[2015]第x030号) 吉林农业科技学院种子基金项目(吉农院合字[2014]第Z07号)
【分类号】：S858.23

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