猪博卡病毒PCR检测方法的建立及初步应用  

Development and preliminary application of the PCR assay for detection of porcine bocavirus

作　　者：周宇[1] 李春萍[2] 朱事康[2] 宋斯伟[3] 徐鹏[4] 陈燕忠[2] 栾维民[1] 

机构地区：[1]吉林农业大学,吉林长春130118 [2]惠州出入境检验检疫局,广东惠州516001 [3]吉林大学动物医学学院,吉林长春130062 [4]辽宁医学院,辽宁锦州121001

出　　处：《中国预防兽医学报》2016年第9期715-718,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家质检总局项目(2014IK243)

摘　　要：为建立能够检测猪博卡病毒（PBoV）所有基因型的检测方法，本研究根据PBoV每个基因群（G1、G2和G3）的保守区域设计特异性引物，建立了检测PBoV的PCR方法。特异性试验结果显示除PBoV外，其它猪病病原核酸扩增均呈阴性，具有较强的特异性；对pMD-GD6、pMD-GD18和pMD-GD4的最低检测限分别为4.64×10^3拷贝/μL、 5.29×10^3拷贝/μL和1.16×10^3拷贝/μL，敏感性较高；采用该方法对39头份猪的肝脏、肺脏、脾脏和小肠内容物进行检测，阳性率为74.4 % （29/39）。其中G1群PBoV的检出率最高，G2群检出率最低；G1群和G3群在4种样品中均有检出，G2群只在小肠内容物中有检出；同时本研究首次在肝脏中检出该病毒。本研究结果表明，PBoV感染情况复杂，存在多基因型的混合感染。本研究为进一步对病毒进行基因分型和致病性研究提供检测依据。In order to establish a method to detect porcine bocavirus （PboV） of all genotypes, a PCR assay was developed with the specific primers designed according to the conserved regions in gene group G1, G2 and G3 of PboV. The test results showed that the PCR assay was specific for PBoV detetion, but had no cross-reaction with other related animal viruses, and the detection limit for recombinant plasmids of pMD-GD6 （group G1）, pMD-GD18 （group G2） and pMD-GD4 （group G3） were 4.64×10^3 copies/μL, 5.29×10^3 copies /μL and 1.16×10^3 copies/μL, respectively. The clinical samples of livers, lungs, spleens and contents of small intestine were from 39 affected pigs and detected by the PCR, and the PboV positive rate was 74.4%. The positive rate of PBoV group G2 was the highest, and the G1 group was the lowest. G1 and G3 were detected in four kinds of samples, but G2 was detected only in the contents of the small intestine. The results showed that PBoV infection was complex and there were mixed infection.

关 键 词：猪博卡病毒 PCR 检测 感染 

分 类 号：S852.65[农业科学—基础兽医学]