猪圆环病毒2型实时荧光RAA检测方法的建立及初步应用

Establishment and Preliminary Application of Fluorescent RAA Detection Method for PCV2

根据猪圆环病毒2型(PCV2)Rep基因序列,设计引物及探针,建立了PCV2实时荧光重组酶介导等温扩增(recombinase-aid amplification, RAA)检测方法。该方法能在40℃恒温条件下20 min内对病原进行快速检测,与猪瘟病毒、伪狂犬病病毒、猪细小病毒、猪传染性胃肠炎病毒、繁殖与呼吸综合征病毒、猪圆环病毒3型和口蹄疫病毒均无交叉反应;对病毒核酸的最低检测限为21.2 copies/μL。用荧光PCR检测方法与建立的荧光RAA检测方法对66份临床样品进行检测,两种方法符合率为90.09%。结果表明,建立的PCV2实时荧光RAA方法适用于PCV2的快速检测。

Primers and probes were designed based on the porcine circovirus type 2(PCV2)Rep gene sequence.A fluorescent RAA detection method for PCV2 was established.This method can quickly detect pathogens within 20 minutes at a constant temperature of 40℃and had no cross-reactivity with CSFV,PRV,PPV,TGEV,PRRSV,PCV3 and FMDV.The minimum detection limit of viral nucleic acid was 21.2 copies/μL.Compared with fluorescent PCR for PCV2,the coincidence rate of the two methods was 90.09%for 66 clinical simples.The results showed that the established RAA method is suitable for the rapid diagnosis of PCV2.