猪圆环病毒3型套式PCR检测方法的建立及应用

Establishment and application of nested PCR for detection of porcine circovirus type 3

为了建立一种准确、快速检测猪圆环病毒3型(PCV-3)的方法,试验根据GenBank中PCV-3基因序列设计合成了内、外两对引物,建立了套式PCR方法,并检验了该方法的特异性、敏感性、重复性,同时用该方法对124份临床疑似病料进行检测。结果表明:用建立的套式PCR方法检测PCV-2、PRV、PPV、PRRSV、CSFV、TGEV、PEDV,均为阴性,外引物PCR扩增的检测灵敏度为17 ng DNA,内引物PCR扩增的检测灵敏度为0. 17 ng DNA。用该方法检测124份临床病料样品,外引物共检测出阳性样品22份,内引物共检测出阳性样品28份,青海省部分地区PCV-3的感染率达22. 58%(28/124)。说明该套式PCR具有良好的特异性、敏感性、重复性和临床适用性,可以用于临床病料中低含量PCV-3的快速检测。

In order to establish an accurate and rapid detection method for porcine circovirus type 3( PCV-3),based on the sequence of PCV-3 gene in GenBank,two pairs of primers were designed and synthesized,and the nested PCR method was established. The specificity,sensitivity and repeatability of the method were tested,and 124 clinical suspected samples were detected by this method.The results showed that PCV-2,PRV,PPV,PRRSV,CSFV,TGEV and PEDV were all negative by nested PCR. The sensitivity of external primer and internal primer was 17 ng DNA and 0. 17 ng DNA respectively. 124 clinical samples were detected by this method,22 positive samples were detected by external primers and 28 positive samples were detected by internal primers. The infection rate of PCV-3 in some areas of Qinghai province was22. 58%( 28/124). It indicats that the nested PCR has good specificity,sensitivity,repeatability,accuracy and clinical applicability,and it can be used for rapid and low-content detection of PCV-3 in clinical materials.