中国生物防治学报2017，Vol.33Issue(6)：796-802,7.DOI:10.16409/j.cnki.2095-039x.2017.06.012
蝗虫微孢子TaqMan探针Real-time PCR检测新方法
Establishment of TaqMan Real-time PCR Method for Detection of Paranosema locustae
摘要
Abstract
Paranosema locustae,an entomopathogen of locusts,is widely used in biocontrol of locusts.The small subunit ribosomal RNA gene of P.locustae was used as the target molecule for designing specific primers and TaqMan probe,pMD18-T,the constructed recombinant plasmid was used to prepare standard agent for fluorescent quantitative PCR amplification.Through optimization of PCR reaction system,linear range of the detection method was determined,and sensitivity and repeatability of the method were evaluated.Finally,a fluorescent quantitative PCR detection method was established successfully.The detectable sensitivity of this method reached 1.29 × 105 copies/L.Three repeated detection trials showed that the variation coefficients within batches and between batches were both less than 1％.These results indicated that the established fluorescent quantitative PCR based on TaqMan probe technology is an accurate,sensitive and easy to operate method for P.locustae detection.关键词
蝗虫微孢子/Real-time PCR/TaqMan探针Key words
Paranosema locustae/fluorescent quantitative PCR/TaqMan probe分类
农业科技引用本文复制引用
扈鸿霞,郑秋英,叶小芳,赵贝,季荣..蝗虫微孢子TaqMan探针Real-time PCR检测新方法[J].中国生物防治学报,2017,33(6):796-802,7.基金项目
新疆维吾尔自治区高校科研计划科学研究重点项目(20140705135643406) （20140705135643406）
新疆维吾尔自治区教育厅重点实验室项目(XJKJT1322-016-01) （XJKJT1322-016-01）
新疆维吾尔自治区天山创新团队计划 （）
