Establishment and preliminary application of nano-PCR method for detecting porcine Kubuvirus
In order to establish a method for rapid diagnosis of porcine kobuvirus and provide a more accurate and convenient method for clinical detection of the virus,this experiment designed a pair of specific primers to amplify the VPO region of the PKV genome.Nano-gold particle materials were added to the traditional PCR reaction system,and after optimization,a convenient,fast and specific nano-PCR method for detecting PKV was established.The test results show that this method can specifi-cally amplify porcine Kubuvirus and has no cross-reactivity with TGEV,PEDV,PDCoV,PRV and PSaV.The nano-PCR showed a higher sensitivity compared to the normal PCR,with a 1 000-fold increased sensitivi-ty.The lowest concentration of nucleic acid detection for ordinary PCR was 5.61 × 10-7 ng/μL,while the lowest nucleic acid detection concentration for nano-PCR was only 5.61 × 10-10 ng/μL.Thirty feces and blood clinical samples from pigs with diarrhea from different pig farms were tested.The results showed that the positive detection rate of nano-PCR was 23.33％(7/30).The positive detection rate of ordinary PCR is 16.66％(5/30).Therefore,the nano-PCR method established in this trial showed higher specifici-ty and sensitivity in the detection of porcine Kubuvirus,which has important clinical significance for the rapid diagnosis and detection of porcine Kubuvirus.
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