摘要
【目的】针对PRRSV Nsp2基因建立反转录重组酶介导等温扩增(RT-RAA)荧光法快速检测方法。【方法】设计经典毒株和高致病毒株引物及探针,检测RT-RAA的敏感性及特异性等。【结果】方法与猪瘟病毒(CSFV)、伪狂犬病毒(PRV Bartha-K61)、猪流行性腹泻病毒(PEDV Purdue)、猪传染性胃肠炎病毒(TGEV LJX)的核酸均无交叉反应;方法对高致病性毒株最低检出限为1.71×10^(1) copies/μL,对经典株最低检出限为2.19×10^(3) copies/μL。高致病毒株方法敏感性为95.5%,特异性为100%;经典株方法敏感性为93.0%,特异性为100%,2种方法具有高度的一致性。【结论】建立的PRRSV RT-RAA荧光法可以区分2种不同的毒株并且特异性良好、灵敏度高。
【Objective】The objective of this study is to establish a rapid detection method for PRRSV Nsp2 gene by reverse transcription recombinase-mediated isothermal amplification(RT-RAA)fluorescence.【Methods】Primers and probes of classical strains and highly virogenic strains were designed to detect the sensitivity and specificity of RT-RAA.【Results】The results showed that the proposed method had no cross-reactivity with the nucleic acids of swine fever virus(CSFV),pseudorabies virus(PRV Bartha-K61),porcine epidemic diarrhea virus(PEDV Purdue)and porcine infectious gastroenteritis virus(TGEV LJX).The minimum detection limit of this method was 1.71×10^(1) copies/μL for highly pathogenic strains and 2.19×10^(3) copies/μL for classical strains.Compared with the PCR method,the sensitivity and specificity of RT-RAA fluorescence quantification method were 95.5% and 100%,respectively.The sensitivity and specificity of RT-RAA classical strain fluorescence quantification method were 93.0%and 100%,respectively and the two methods were highly consistent.【Conclusion】The PRRSV RT-RAA fluorescence method established in this study can distinguish two different strains with good specificity and high sensitivity.
作者
于娜
赵爱云
黄春媛
马佳镁
张紫薇
范悦轩
郑佳馨
张艳
刘光亮
齐萌
曹宗喜
YU Na;ZHAO Aiyun;HUANG Chunyuan;MA Jiamei;ZHANG Ziwei;FAN Yuexuan;ZHENG Jiaxin;ZHANG Yan;LIU Guangliang;QI Meng;CAO Zongxi(Tarim Animal Disease Diagnosis and Prevention Engineering Laboratory of Xinjiang Production and Construction Corps/College of Animal Science and Technology,Tarim University,Aral Xinjiang 843300,China;Hainan Provincial Key Laboratory of Tropical Animal Breeding and Disease Research,Institute of Animal Science and Veterinary Medicine/Hainan Academy of Agricultural Sciences,Haikou 571100,China;College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China;Suzhou Mars Biotechnology Co,Ltd.,Suzhou Jiangsu 215021,China;Hainan Experimental Animal Research Center,Sanya Research Institute,Hainan Academy of Agricultural Sciences,Sanya Hainan 572000,China)
出处
《新疆农业科学》
北大核心
2025年第3期748-753,共6页
Xinjiang Agricultural Sciences
基金
国家重点研发计划项目(2023YFC3404302,2021YFA0805905)
海南省重点研发计划项目(ZDYF2023XDNY038)
国家自然科学基金项目(32060796)
海南省重大科技计划项目(ZDKJ2021030)
