当前位置: 首页 > 文章 > 美洲幼虫腐臭病二温式PCR诊断方法的建立及应用 新疆农业科学 2015 (2) 288-291
Position: Home > Articles > Establishment of a Two- temperature PCR Assay for Detection of American Foulbrood Xinjiang Agricultural Sciences 2015 (2) 288-291

美洲幼虫腐臭病二温式PCR诊断方法的建立及应用

作  者:
何晓杰;阿曼吐尔·阿黑哈提;叶尔保勒;史秀丽;齐鑫;皮志媛;王振宝
单  位:
伊犁职业技术学院;新疆维吾尔自治区蜂业发展中心;伊犁出入境检验检疫局
关键词:
美洲幼虫腐臭病;二温式PCR;建立
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。
译  名:
Establishment of a Two- temperature PCR Assay for Detection of American Foulbrood
作  者:
HE Xiao-jie;Amantuer Aheihati;Yeerbaole;SHI Xiu-li;QI Xin;PI Zhi-yuan;WANG Zhen-bao;Yili Vocational and Technical College;Yili Entry - Exit Inspection and Quarantine Bureau;Apiculture Development Center,Xinjiang Uygur Autonomous Region;
关键词:
American foulbrood;;two-temperature PCR;;establishment
摘  要:
【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.

相似文章

计量
文章访问数: 8
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊