Microfluidic Chip Loop-mediated Isothermal Amplification for Detecting Three Porcine Circoviruses
In order to rapidly distinguish three types of porcine circoviruses(PCV2,PCV3 and PCV4)at field level,clinical positive samples infected with the three circoviruses were extracted for nucleic acids for microfluidic chip loop-mediated isothermal amplification,the results of which were synchronously compared with those of three commercial fluorescent probe kits in the aspects of sensitivity,specificity and repeatability.The results indicated that the microfluidic chip loop-mediated isothermal amplification method was highly sensitive in joint-detection of the three viruses,with the sensitivity no lower than fluorescent quantification PCR within 30 minutes,failed to crossly react with clinical positive samples infected with African swine fever virus(ASFV),classical swine fever virus(CSFV),porcine reproductive and respiratory syndrome virus(PRRSV),porcine pseudorabies virus(PRV)and porcine parvovirus(PPV),indicating good specificity;and all the coefficients of variation(CVs)of the repeated Ct values after detection of the three viruses were below 2%,showing good stability.In conclusion,the microfluidic chip method for joint-detection of the three viruses was characterized with the advantages of good specificity,high sensitivity,strong repeatability,rapid detection and low requirements for environment,and appropriate for rapid on-site detection of porcine circoviruses on swine farms and relevant premises.The established method could be used as a strong tool for rapid on-site detection of relevant pig pathogens.
porcine circovirusmicrofluidic chiploop-mediated isothermal amplificationrapid detectionnucleic acid test
国家科技期刊平台
NSTL
万方数据
维普
