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以色列急性麻痹病毒(IAPV)RT-PCR检测方法的建立及应用

Establishment and application of RT-PCR method for detection of IAPV
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摘要 为建立以色列急性麻痹病毒(Israeli acute paralysis virus,IAPV)特异性核酸检测方法,本研究以IAPV polvmerase polyprotein基因为靶基因设计合成1对特异性引物,建立了IAPV的RTPCR检测方法。结果显示,对蜜蜂的5种不同病毒进行RT-PCR检测,只有IAPV病毒检测结果为阳性,具有良好的特异性;对IAPV的最低检测限为10^3拷贝/μL,具有较高的灵敏度;利用该检测方法对来自福建省不同养蜂场的8份蜜蜂样品进行检测,3份样品为阳性,通过序列比对分析,同源性超过99%,进一步证实为IAPV。结果表明,所建立的IAPV RT-PCR检测方法灵敏度高、特异性强、重复性好,具有良好的实用性。 To establish a rapid assay for Israeli acute paralysis virus(IAPV), RT-PCR method was developed targeting polymerase polyprotein gene of IAPV. The results showed that the test for 5 different viruses of bees with RT-PCR method, only IAPV test was positive, indicating that the method had high specificity. In addition, the sensitivity of RT-PCR was 103 copies/μL. Furthermore,a total 3 positive samples for IAPV were detected from 8 bees samples of different apiary by the RT-PCR method, at the same time, sequence alignment analysis showed the homology was more than 99%, which showed that the method had good practicability. Therefore, the RT-PCR method provided a novel rapid, sensitive and good repeatability detection method for IAPV infection.
作者 张体银 李丹丹 郑腾 白泉阳 张志灯 王武军 于师宇 ZHANG Ti-yin1 , LI Dan-dan2 , ZHENG Teng1 , BAI Quan-yang1 , ZHANG Zhi-deng1 , WANG Wu-jun1 ,YU Shi-yu1 (1. Fujian Provincial Key Laboratory of Inspection and Quarantine Technology Research, Fujian Entry-Exit Inspection and Quarantine Bureau, Fuzhou 350001, China;2. Technical Center of Hainan Entry-Exit Inspection and Quarantine Bureau, Haikou 570311, China)
出处 《中国兽医学报》 CSCD 北大核心 2018年第6期1105-1108,共4页 Chinese Journal of Veterinary Science
基金 福建省科技计划重点资助项目(2014N0001) 福建检疫局科技计划资助项目(FK2015-JS001) 海南省社会发展科技专项资助项目(2015SF29)
关键词 以色列急性麻痹病毒 RT-PCR POLYMERASE polyprotein基因 蜜蜂 IAPV RT-PCR ploymerase ployprotein gene bee
作者简介 张体银(1979-),男,高级工程师,硕士。通讯作者,E-mail:zhty5337@163.com
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