Development and Application of Triple PCR for Detection and Differentiation of PCV-2,PCV-3 and PCV-4
To develop a rapid method for detection and differentiation of PCV-2,PCV-3 and PCV-4,the specific primers of PCV-2,PCV-3 and PCV-4 were designed and synthesized,for a triple PCR method.The specific bands of 216,415,678,216 bp/415 bp/678 bp were amplified from PCV-2,PCV-3,PCV-4 and PCV-2/PCV-3/PCV-4.The triple PCR method did not amplify PRV,PPV,PRRSV,CSFV,PEDV and TGEV.The minimum detection limits were 1.0×102 copies/µL for PCV-2,1.0×103 copies/µL for PCV-3 and 1.0×103 copies/µL for PCV-4.The coincidence rates of the single PCR method for PCV-2,PCV-3 and PCV-4 were all 100%.Then,85 clinical samples were tested by this method and the positive rates of PCV-2,PCV-3 and PCV-4 were 21.17%,14.12%and 4.71%,respectively.These results showed that the developed triple PCR method for differential detection of PCV-2,PCV-3 and PCV-4 had good specificity,sensitivity,accuracy and clinical applicability,thus provided a simple and rapid molecular biological diagnostic technology for PCV-2,PCV-3 and PCV-4.
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