西花蓟马qRT-PCR与常规PCR快速检测比较  

Comparation of real-time fluorescent quantitative PCR and normal PCR for the rapid identification of Frankliniella occidentalis

作　　者：汤云霞[1] 严丹侃[1,2] 范加勤[1] 

机构地区：[1]南京农业大学植物保护学院,江苏南京210095 [2]安徽省农业科学院

出　　处：《植物检疫》2012年第6期23-26,共4页Plant Quarantine

基　　金：国家公益性行业(农业)科研专项经费项目(201103026)

摘　　要：西花蓟马是一种世界性害虫,其体型小、隐匿性强,因形态学鉴定必需成虫,存在不足。为此本研究以西花蓟马线粒体DNA的COI基因为模板,设计了1对引物,建立了西花蓟马qRT-PCR检测方法。该引物具有特异性,只有西花蓟马的荧光信号能被检测;检测灵敏度达到1/10000头成虫,靶标DNA片段的稀释限为11.63拷贝/μL,而常规PCR法的检测限点为1/120头成虫。该检测体系与常规PCR相比,灵敏度高、快速准确,对进出口检验检疫以及产品调运中入侵物种的检测具有重要意义。The western flower thrips （Frankliniella occidentalis） is an important pest in agriculrue worldwidely. For the small body type and secluded behaviour of this pest, the methods of traditional morphological identification cannot do anything with thrlps in nonaduh stages. In this study, based on the COI gene of mtDNA in F. occidentalis, we designed a pair of specific primers, and set up a rapid detection method for the western flower thrips using real - time fluorescent quantitative PCR. The results showed that only the fluorescence signal of western flower thrips could be detected by this pair of primers, with which 1/10000 of this thrips adult was be catched, and the content of target DNA is 11.63 copies/μL. But the normal PCR could only detect 1/120 adult of this pest. In conclusion, this detection system has more high sensitivity than normal PCR method, it is important in the inspection and quarantine and the detections of invasive species during transportation of products.

关 键 词：西花蓟马 实时荧光定量PCR 常规PCR 检测 

分 类 号：S433[农业科学—农业昆虫与害虫防治]