微流控芯片法在非洲猪瘟病毒核酸快速检测中的性能分析

Performance of the microfluidic chip in rapid detection of African swine fever virus nucleic acid

为评估微流控芯片法在非洲猪瘟病毒(ASFV)核酸快速检测中的各项性能,验证该方法的敏感性、特异性、重复性和临床效果,本研究将ASFV VP72质粒标准物质、猪瘟病毒(CSFV)、猪流行性腹泻病毒(PEDV)、猪传染性胃肠炎病毒(TGEV)、猪圆环病毒2型(PCV2)和伪狂犬病病毒(PRV)培养物各1份以及临床样本43份进行核酸提取,用ASFV微流控芯片快速检测试剂盒进行测试研究,并与市场上某品牌ASFV荧光定量PCR检测试剂盒进行比对。结果表明:微流控芯片法可以在15~30 min内,实现2.5 copies/μL的最低检出限,与荧光定量PCR具有相同的敏感性和重复性;同时该方法对CSFV、PEDV、TGEV、PCV2和PRV病毒培养物均无交叉反应,特异性良好;通过对43份临床样本测试显示,微流控芯片法与荧光定量PCR的临床符合率为97.67%(42/43)。研究表明,微流控芯片法在ASFV核酸检测上性能良好。

In order to evaluate the performance of the microfluidic chip in rapid detection of African swine fever virus(ASFV)nucleic acid,the sensitivity,repeatability,specificity and clinical effect of the method were analyzed in this study.One share of culture for each of the nucleic acids of the ASFV VP72 plasmid reference material,classical swine fever virus(CSFV),porcine epidemic diarrhea virus(PEDV),transmissible gastroenteritis virus(TGEV),porcine circovirus type 2(PCV2),pseudorabies virus(PRV)and 43 corresponding clinical samples were extracted,and tested with an ASFV microfluidic chip rapid detection kit,and the tested results were compared with a commercial ASFV fluorescence quantitative PCR detection kit.The results showed that the detection was finished within 15-30 minutes and the limitation of the microfluidic chip was 2.5 copies/μL,which was of the same sensitivity and repeatability as those of the fluorescence quantitative PCR.Meanwhile,the specificity of the studied method was good,and there was no cross reaction to CSFV,PEDV,TGEV,PCV2 and PRV virus cultures in the test.The results of the 43 clinical samples showed that the clinical coincidence rate of the microfluidic chip method with the fluorescent quantitative PCR was 97.67%(42/43).It suggests that microfluidic chip has a good performance in detection of ASFV nucleic acid.