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【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。
【Objective】 The objective of this study was to design a real-time PCR method for the rapid and sensitive detection of the fruit flies in the Mediterranean. 【Method】 The DNA fragments of the fruit fly and the other three fruit flies were amplified by the universal primers of mitochondrial DNA COI gene of fruit flies, and the specific primers were designed to amplify the fruit flies , SYBR Green real-time PCR method to verify the specificity and sensitivity of the primer. 【Result】 The designed primer pairs DZH-F / DZH-R were able to detect the Mediterranean fruit flies. The peak Tm of the amplification curve was 79.2. The sensitivity of the pair of primers for detection of Mediterranean fruit flies was 10-3 ng / μL. 【Conclusion】 SYBR Green real-time fluorescence PCR method was established to detect Mediterranean fruit fly with high specificity and high sensitivity.