当前位置: 首页 > 文章 > 实时荧光PCR法快速检测地中海实蝇的研究 新疆农业科学 2016,53 (6) 1107-1113
Position: Home > Articles > Study on Rapid Detection Method of Mediterranean Fruit Fly by Real- Time PCR Assay Xinjiang Agricultural Sciences 2016,53 (6) 1107-1113

实时荧光PCR法快速检测地中海实蝇的研究

作  者:
李京;张祥林;罗明;李亚伟;王翀;张小菊
单  位:
新疆出入境检验检疫局;新疆农业大学农学院
关键词:
地中海实蝇;COI基因;实时荧光PCR;快速检测
摘  要:
【目的】设计用于扩增地中海实蝇的特异性引物,建立快速、灵敏检测地中海实蝇的实时荧光PCR方法。【方法】采用实蝇科昆虫线粒体DNA COI基因的通用引物扩增地中海实蝇以及其它三种供试实蝇的DNA片段,经测序、比对分析,设计出扩增地中海实蝇的特异性引物,用SYBR Green实时荧光PCR方法验证该引物的特异性与灵敏度。【结果】设计的引物对DZH-F/DZH-R能特异性检测出地中海实蝇,扩增产物的溶解曲线峰值Tm为79.2。该对引物检测地中海实蝇的灵敏度为10-3ng/μL。【结论】所建立的SYBR Green实时荧光PCR方法检测地中海实蝇特异性强、灵敏度高。
译  名:
Study on Rapid Detection Method of Mediterranean Fruit Fly by Real- Time PCR Assay
作  者:
LI Jing;ZHANG Xiang-lin;LUO Ming;WANG Chong;LI Ya-wei;ZHANG Xiao-ju;College of Agronomy,Xinjiang Agricultural University;Xinjiang Entry and Exit Inspection and Quarantine Bureau;
关键词:
Mediterranean fruit fly;;mtDNA;;SYBR Green real-time PCR;;rapid detection
摘  要:
【Objective】Fruit flies( Tephritidae) described all over the world are more than 450 insect species and 4,300 kinds,there were more than 70 kinds including the Mediterranean fruit fly( Ceratitis capitate) in Xinjiang,which endanger agriculture greatly. It is of significance in plant quarantine to set up quarantine fruit fly,especially immature worm state fast and reliable identification method. 【Method】Using the fruit flies general primer amplification of mitochondrial DNA COI gene to amplify targeted fragments,sequence,blast,then design specific primers which verified its specificity and sensitivity using SYBR Green real- time PCR.【Result】The primers DZH- F / DZH- R could specific amplify ceratitis capitate. The production peaks was 79. 2. This primer could detect 10- 3ng / μL of ceratitis capitate template DNA.【Conclusion】A assay of SYBR green real- time PCR has been established to detect the Mediterranean fruit fly,which has strong specificity and high sensitivity.

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