猪博卡病毒 N P1 基因特异性的PCR 方法建立及应用

叶星宇,聂奎,聂福平

doi:10.13718/j.cnki.xdzk.2015.05.003

西南大学学报（自然科学版）

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猪博卡病毒 N P1 基因特异性的PCR 方法建立及应用

叶星宇,聂奎,聂福平. 猪博卡病毒 N P1 基因特异性的PCR 方法建立及应用[J]. 西南大学学报（自然科学版）, 2015, 37(5). doi: 10.13718/j.cnki.xdzk.2015.05.003

引用本文:

叶星宇,聂奎,聂福平. 猪博卡病毒 N P1 基因特异性的PCR 方法建立及应用[J]. 西南大学学报（自然科学版）, 2015, 37(5). doi: 10.13718/j.cnki.xdzk.2015.05.003

YE Xing-yu,NIE Kui,NIE Fu-ping. Establishment of a PCR Assay Specifically for PBoV Based on NP1 and Its Preliminary Application[J]. Journal of Southwest University Natural Science Edition, 2015, 37(5). doi: 10.13718/j.cnki.xdzk.2015.05.003

Citation:

YE Xing-yu,NIE Kui,NIE Fu-ping. Establishment of a PCR Assay Specifically for PBoV Based on NP1 and Its Preliminary Application[J]. Journal of Southwest University Natural Science Edition, 2015, 37(5). doi: 10.13718/j.cnki.xdzk.2015.05.003

猪博卡病毒 N P1 基因特异性的PCR 方法建立及应用

叶星宇,聂奎,聂福平

西南大学动物科技学院 ,重庆400715;四川省广元市动物疫病预防控制中心 ,四川广元628000 西南大学动物科技学院 ,重庆,400715 重庆市出入境检验检疫局 ,重庆,400020

Establishment of a PCR Assay Specifically for PBoV Based on NP1 and Its Preliminary Application

YE Xing-yu,NIE Kui,NIE Fu-ping

摘要: 该研究根据Genbank公布的猪博卡病毒Porcine bocavirus ,PBoV的核苷酸序列 ,在其NP1保守区域设计一对特异性引物 ,建立一种能扩增多种猪博卡病毒亚型的PCR检测方法 ,并进行特异性试验 ,敏感性试验 ,重复性试验及可靠性检测 .结果显示 :所建立的PCR检测方法对猪瘟病毒、猪细小病毒、猪繁殖与呼吸综合征病毒、猪圆环病毒2型无交叉反应 ;敏感度可以检测到58 pg/μL ;经3次重复检测 ,结果一致 ,重复性好 ;对阳性产物测序 ,与Genbank公布的猪博卡病毒NP1序列同源性在97.3% 以上 ,可靠性强 .用建立的PCR方法对2011年重庆部分地区猪场的266份育肥猪血清样本进行检测 ,结果45份为猪博卡病毒阳性 ,证明了在重庆地区育肥猪中存在PBoV感染的情况 .
- 猪博卡病毒 /
- NP1基因 /
- PCR检测方法
Abstract: According to the published PBoV nucleotide sequence in GenBank ,we designed a pair of specific primers in the conserved region of NP1 to establish a PCR method that can amplify a variety of PBoV sub-types and tested its specificity ,sensibility ,repeatability and reliability .The results showed that the meth-od had no cross reaction for classical swine fever virus ,porcine parvovirus ,porcine reproductive and re-spiratory syndrome virus and porcine circovirus type II ,indicating that its specificity is high .Its sensibility was high ,with 58 ng ,and its repeatablity was good ,as the results were consistent in 3 repeative tests .We sequenced the positive products and the similarity with PBoV-NP1 published by GenBank was more than 97.3% ,suggesting that the method was reliable .We applied the established method to test 266 pig serum samples collected in Chongqing area in 2011 .Forty-five samples were shown to contain PBoV ,proving that the infection of PBoV existed among fattening pigs in Chongqing area .
- Porcine bocavirus (PBoV)，NP1 gene，PCR method /

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猪博卡病毒 N P1 基因特异性的PCR 方法建立及应用

叶星宇,聂奎,聂福平

西南大学动物科技学院 ,重庆400715;四川省广元市动物疫病预防控制中心 ,四川广元628000 西南大学动物科技学院 ,重庆,400715 重庆市出入境检验检疫局 ,重庆,400020

关键词:

摘要: 该研究根据Genbank公布的猪博卡病毒Porcine bocavirus ,PBoV的核苷酸序列 ,在其NP1保守区域设计一对特异性引物 ,建立一种能扩增多种猪博卡病毒亚型的PCR检测方法 ,并进行特异性试验 ,敏感性试验 ,重复性试验及可靠性检测 .结果显示 :所建立的PCR检测方法对猪瘟病毒、猪细小病毒、猪繁殖与呼吸综合征病毒、猪圆环病毒2型无交叉反应 ;敏感度可以检测到58 pg/μL ;经3次重复检测 ,结果一致 ,重复性好 ;对阳性产物测序 ,与Genbank公布的猪博卡病毒NP1序列同源性在97.3% 以上 ,可靠性强 .用建立的PCR方法对2011年重庆部分地区猪场的266份育肥猪血清样本进行检测 ,结果45份为猪博卡病毒阳性 ,证明了在重庆地区育肥猪中存在PBoV感染的情况 .

English Abstract

Establishment of a PCR Assay Specifically for PBoV Based on NP1 and Its Preliminary Application

西南大学动物科技学院 ,重庆400715;四川省广元市动物疫病预防控制中心 ,四川广元628000 西南大学动物科技学院 ,重庆,400715 重庆市出入境检验检疫局 ,重庆,400020

Keywords:

Abstract: According to the published PBoV nucleotide sequence in GenBank ,we designed a pair of specific primers in the conserved region of NP1 to establish a PCR method that can amplify a variety of PBoV sub-types and tested its specificity ,sensibility ,repeatability and reliability .The results showed that the meth-od had no cross reaction for classical swine fever virus ,porcine parvovirus ,porcine reproductive and re-spiratory syndrome virus and porcine circovirus type II ,indicating that its specificity is high .Its sensibility was high ,with 58 ng ,and its repeatablity was good ,as the results were consistent in 3 repeative tests .We sequenced the positive products and the similarity with PBoV-NP1 published by GenBank was more than 97.3% ,suggesting that the method was reliable .We applied the established method to test 266 pig serum samples collected in Chongqing area in 2011 .Forty-five samples were shown to contain PBoV ,proving that the infection of PBoV existed among fattening pigs in Chongqing area .