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在设计的枣实蝇15对引物中,利用种特异引物PCR(SS-PCR)和琼脂糖凝胶电泳技术筛选出1对枣实蝇的特异性引物,引物的特异性用桔小实蝇、瓜实蝇、南瓜实蝇、番石榴实蝇和桃果实蝇等5种实蝇来验证。SS-PCR方法的检测灵敏度用40,20,10,1,0.1,0.01,0.001 ng·μL-1等7个不同浓度系列的枣实蝇DNA模板来检测。结果表明,SS-PCR方法的检测限度达0.01 ng·μL-1以下,最适模板DNA浓度为1~20 ng·μL-1。所设计的引物对枣实蝇不同虫态均能进行特异性扩增,结果准确可靠。该技术体系可用于枣实蝇的鉴定识别与检测监测,对阻止其进一步扩散蔓延具有重要意义。
A total of 15 pairs of primers were designed to identify the specific primers of fruit jujube, using specific primers PCR (SS-PCR) and agarose gel electrophoresis, Melon fruit fly, pumpkin fruit fly, guava fruit fly and peach fruit flies and other 5 kinds of fruit flies to verify. The detection sensitivity of SS-PCR method was tested with DNA template of jujube fruit fly at the concentrations of 40, 20, 10, 1, 0.1, 0.01 and 0.001 ng · μL-1. The results showed that the detection limit of SS-PCR method was less than 0.01 ng · μL-1, and the optimal template DNA concentration was 1 ~ 20 ng · μL-1. The designed primers can specifically amplify the different insect states of jujube fruit fly with accurate and reliable results. The technology system can be used for identifying, detecting, monitoring and monitoring Jujube fruit fly, and is of great significance for preventing further spread of Jujube fruit fly.