中国动物传染病学报2018，Vol.26Issue(1)：25-31,7.
环介导等温扩增技术与横向流动试纸条法快速检测鸡贫血病毒的研究
RAPID AND SENSITIVE DETECTION OF CHICKEN ANEMIA VIRUS BY LOOP-MEDIATED ISOTHERMAL AMPLIFICATION COMBINED WITH LATERAL-FLOW DIPSTICK
李秀梅 1石瑜 1朱雅宁 1袁雪涛 1杨爱华 1杨颖1
作者信息
- 1. 天津市动物疫病预防控制中心,天津 300402
- 折叠
摘要
Abstract
In the present study, we established a rapid Chicken anemia virus (CAV) detection method by combining loop-mediated isothermal amplification (LAMP) with lateral flow dipstick (LFD) examination. The method utilized four specific primers targeting CAV Cux-1 VP2 gene for LAMP and fluorescein isothiocyanate (FITC)-labeled probe to specifically hybrid to the biotin-labeled LAMP product for LFD examination. The optimized amplification temperature and time were 63℃ and 50 min, respectively and whole procedure time was only 80 min including sample process. The LAMP-LFD method developed in this study specifically detected CAV and showed no reaction with other 6 immunosuppressive viruses. The detection sensitivity was 10 fg, i.e. 100-fold lower than that of conventional PCR using the outer primer set. This method had good repeatability and stability. The LAMP-LFD method was used to test 30 specimens and compared with virus isolation and identification method. As a result, both methods had 100% agreement. Overall, the LAMP-LFD method rapidly and specifically detected CAV and did not required other auxiliary equipment, thus suitable for basic laboratory testing, emergency use and on-site monitoring.关键词
鸡贫血病毒/VP2基因/环介导等温扩增/横向流动试纸条/检测Key words
Chicken anemia virus/VP2 gene/loop-mediated isothermal amplification/lateral flow dipstick test/detection分类
农业科技引用本文复制引用
李秀梅,石瑜,朱雅宁,袁雪涛,杨爱华,杨颖..环介导等温扩增技术与横向流动试纸条法快速检测鸡贫血病毒的研究[J].中国动物传染病学报,2018,26(1):25-31,7.
