畜牧兽医学报2024，Vol.55Issue(1)：406-412,7.DOI:10.11843/j.issn.0366-6964.2024.01.039
牛诺瓦病毒和牛轮状病毒双重RAA-LFD快速检测方法的建立及应用
Establishment and Application of Rapid Detection Method for Bovine Norovirus and Bovine Rotavirus Dual RAA-LFD
摘要
Abstract
This study aimed to establish a recombinase aided amplification-lateral flow dipstick(RAA-LFD)method for simultaneous detection of bovine norovirus(BNoV)and Bovine rotavirus(BRV).According to the principle of RAA primer probe design,this test first designed primers and probes targeted the conserved regions of BNoV RdRp gene and BRV VP7 gene,second prepared the standard recombinant plasmids,then established and optimized the RAA-LFD reaction system,finally the specificity,sensitivity and repeatability of the detection method were also evaluated.At the same time,168 clinical samples of calves with diarrhea were tested in parallel by the RAA-LFD,PCR and qPCR method.The results showed that the method could amplify the target fragment in 20 minutes at 39 ℃.The sensitivity of this method is 103 copies·μL-1 for both BNoV and BRV,and is basically consistent with the detection results of PCR.It has no cross-reaction with bovine coronavirus(BCoV),bovine viral diarrhea virus(BVDV)and infectious bovine rhinotracheitis virus(IBRV).Although the test strip result may be lower than that of the qPCR test,it can be easily observed with the naked eye.In summary,the RAA-LFD method has high sensitivity,and is simple,fast,and does not require specialized equipment or operators,making it ideal for on-site detection.关键词
牛诺瓦病毒/牛轮状病毒/重组酶介导扩增/侧向流试纸条Key words
bovine norovirus/bovine rotavirus/recombinant enzyme-mediated amplification/lateral flow test strips分类
农业科技引用本文复制引用
李惠惠,董可儿,刘馨博,张春晓,马超,陈利苹,钟旗,姚刚,马雪连..牛诺瓦病毒和牛轮状病毒双重RAA-LFD快速检测方法的建立及应用[J].畜牧兽医学报,2024,55(1):406-412,7.基金项目
国家自然科学基金(32002251) （32002251）
肉生高效健康饲养技术集成研究与示范(2021A02003-1) （2021A02003-1）
