美洲幼虫腐臭病二温式PCR诊断方法的建立及应用  

Establishment of a Two- temperature PCR Assay for Detection of American Foulbrood

作　　者：何晓杰[1] 阿曼吐尔.阿黑哈提 叶尔保勒 史秀丽 齐鑫 皮志媛[1] 王振宝 

机构地区：[1]伊犁职业技术学院,新疆伊宁835000 [2]伊犁出入境检验检疫局,新疆伊宁835000 [3]新疆维吾尔自治区蜂业发展中心,乌鲁木齐830054

出　　处：《新疆农业科学》2015年第2期288-291,共4页Xinjiang Agricultural Sciences

基　　金：国家质检总局科技计划项目(2012IK023);伊犁职业技术学院科研计划项目(yzyxm2012003);新疆维吾尔自治区青年科学基金项目(201318101-19)

摘　　要：【目的】为建立蜂及蜂产品中美洲幼虫腐臭病(AFB)二温式PCR诊断方法。【方法】选取幼虫芽孢杆菌基因保守序列设计一对特异性引物,优化反应参数,进行特异性、敏感性和重复性试验验证,建立了诊断AFB的二温式PCR方法。【结果】扩增出130 bp的特异性片段,与参考株序列相似性为100%,DNA灵敏度检测幼虫芽孢杆菌达到33 fg/PCR体系,重复性良好。应用该方法对40份实验室模拟样本和50份临床样本进行了检测,与预期结果一致。【结论】该方法具有特异、灵敏、准确等优点,可用于蜂及蜂产品中AFB的快速诊断。【Objective】To establish a method for American foulbrood detection in bees and bee products by using two- temperature PCR.【Method】A pair of primers was designed according to the conservative gene sequences of Paenibacillus larvae.【Result】Under the optimized reaction parameter,experiments verified that the PCR assay was highly specific to amplify a 130 bp fragment from Paenibacillus larvae genomic DNA with a detection limit of 33 fg / PCR and good repeatability. The two- temperature PCR was applied to detect 40 simulation samples and 50 clinical samples,the result was consistent with the prediction in the theory. 【Conclusion】The result showed that the two- temperature PCR technique provided a useful method for American foulbrood detection in bees and bee products.

关 键 词：美洲幼虫腐臭病 二温式PCR 建立 

分 类 号：S89[农业科学—特种经济动物饲养]