为建立同时检测猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus，PRRSV)、猪流感病毒(Swine influenza virus，SIV)、伪狂犬病病毒(Pseudorabies virus，PRV)和猪圆环病毒2 型(Porcine circovirus 2，PCV2)4种病毒的多重PCR，分别针对PRRSV、SIV、PCV2 和PRV 保守序列设计4对特异性引物，建立同时检测这4种病毒的多重PCR，并对采自呼吸障碍病猪的病料进行检测。结果显示，PRRSV、SIV、PCV2 、PRV 扩增产物分别是275，650，482，359 bp，检测最小DNA量分别是1.5，7.5，4.3，2.7 pg；检测采自河南省的100份样品，总阳性率为95%(95/100)，其中PRRSV、SIV、PCV2 和PRV 阳性率分别是62.1%(59/95)，33.7%(32/95)，66.3%(63/95)，26.3%(25/95)，混合感染率达69.5%(66/95)。表明建立的多重PCR方法敏感、特异、快速，可用于猪呼吸道疾病综合征常见病毒的检测。
赵朴
,
郑玉姝
,
赵坤
,
李任峰
,
胡建和
,
王三虎
,
刘兴友
,
陈金山
,
刘俊伟
. 猪呼吸道疾病综合征常见病毒多重PCR方法的建立及应用[J]. 华北农学报, 2014
, 29(3)
: 64
-67
.
DOI: 10.7668/hbnxb.2014.03.013
To detect porcine reproductive and respiratory syndrome virus(PRRSV), swine influenza virus(SIV), pseudorabies virus(PRV) and porcine circovirus type 2(PCV2) simultaneously, four pairs of primers specific to PRRSV, SIV, PCV2 and PRV are designed and a multiplex PCR was developed.The samples from pigs of respiratory failure were detected by the multiplex PCR.The results showed that product for each virus produce four distinct fragments 275, 650, 482 and 359 bp, representing PRRSV, SIV, PCV2 and PRV, respectively.The DNA detection limit of the multiplex PCR was 1.5, 7.5, 4.3 and 2.7 pg for PRRSV, SIV, PCV2 and PRV, respectively.Among 100 pig specimens from Henan province, the total positive rate was 95%(95/100) and within them, the positive rate for PRRSV, SIV, PCV2 and PRV was 62.1%(59/95), 33.7%(32/95), 66.3%(63/95) and 26.3%(25/95) respectively.And the co-infection rate was 69.5%(66/95).This multiplex PCR is sensitive, specific and rapid method for the detection and identification of major viruses involved in porcine respiratory disease complex.
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