鸽新城疫病毒可视化LAMP检测方法的建立与应用

Establishment and Application of Visualization LAMP for Detection of Pigeon Newcastle Disease Virus

为建立一种简便、快速,适用于基层检测鸽Ⅰ型副黏病毒的方法,研究针对鸽Ⅰ型副黏病毒全基因组保守序列,设计了1组可检测基因Ⅵ、Ⅶ、ⅩⅩ、ⅩⅪ型分离株的环介导等温扩增(LAMP)引物,通过优化反应条件及体系,建立鸽Ⅰ型副黏病毒可视化LAMP检测方法。结果显示:建立的鸽Ⅰ型副黏病毒可视化LAMP检测方法可检测基因Ⅵ、Ⅶ、ⅩⅩ、ⅩⅪ型鸽新城疫病毒,与H9亚型禽流感病毒、马立克病病毒、禽网状内皮组织增殖病病毒、小鹅瘟病毒以及禽白血病病毒以及新城疫病毒La Sota疫苗株均无交叉反应,最低检测限为10^(1)copies/μL。研究表明所建立的鸽Ⅰ型副黏病毒可视化LAMP检测方法快速、简便,特异性强,灵敏度高,可应用于基层快速检测鸽Ⅰ型副黏病毒。

To establish a simple and rapid method for the detection of pigeon paramyxovirus serotype-1 in pigeons at the grass-roots level,a set of reverse transcription loop-mediated isothermal amplification(RT-LAMP)primers were designed ac⁃cording to the conservative sequences of pigeon paramyxovirus serotype-1 whole genome,includingⅥ,Ⅶ,ⅩⅩandⅩⅪgeno⁃type isolated strains.The visual LAMP detection method was developed by optimizing the reaction conditions and system.The re⁃sults showed that the established visual LAMP detection method could detectⅥ,Ⅶ,ⅩⅩ,andⅩⅪgenotype pigeon paramyxo⁃virus,and had no cross reactivity with subtype H9 avian influenza virus,Marek's disease virus,reticuloendotheliosis virus,gos⁃ling plague virus,avian leukosis virus and La Sota strain of Newcastle disease virus.Besides,the minimum detection level was 10^(1)copies/μL.The visual LAMP detection method established in this study was proved to be fast,simple,specific,and sensitive and which could play an important role in the rapid detection of Newcastle disease virusDV in pigeons at the grassroots level.