非洲猪瘟病毒NanoPCR检测方法的建立

Establishment of NanoPCR detection method for African swine fever virus

为建立一种快速、灵敏、适用于大批量样品检测非洲猪瘟病毒(ASFV)的方法,本研究使用金纳米颗粒与传统PCR技术相结合,针对ASFV中P72保守序列,建立了一种新的ASFV的NanoPCR分子检测方法,并对其特异性和敏感性进行了研究。结果表明,对于相关病毒,包括猪繁殖与呼吸综合征病毒(PRRSV)、猪圆环病毒2型(PCV2)和猪瘟病毒(CSFV),NanoPCR分析均未发现交叉反应。与常规PCR相比,NanoPCR灵敏度更高,每个反应的检出限为2.86×10~3copies/μL,而常规PCR检出限为2.86×10~4copies/μL。用建立的方法对23份临床样品进行检测,结果 ASFV的阳性检出率为60.86%。综上所述,本研究开发的灵敏、特异性的NanoPCR方法可广泛应用于ASFV感染的临床诊断和现场监测。

In order to establish a rapid,sensitive,and suitable method for the detection of African swine fever virus(ASFV) in large quantities of samples,this study uses gold nanoparticles combined with traditional PCR technology to establish a new method for the conservative sequence of P72 in ASFV.Nano PCR molecular detection method of ASFV,and its specificity and sensitivity have been studied.The results showed that the Nano PCR has good specificity and did not cross-react with porcine reproductive and respiratory syndrome virus,porcine circovirus 2,infection with classical swine fever virus.Compared with conventional PCR,Nano PCR has higher sensitivity.The detection limit of each reaction is 2.86×10^3 copies/μL,while the detection limit of conventional PCR analysis is 2.86×10^4 copies/μL.Using the method to detect 23 clinical samples,the positive rate of ASFV was 60.86%.In summary,the sensitive and specific Nano PCR detection method developed in this research can be widely used in clinical diagnosis and field monitoring of ASFV infection.