摘要
为建立快速检测猪博卡病毒G1基因群(PBoV G1)的PCR方法,参照GenBank中最新收录的PBoV G1的VP1基因保守序列,设计了1对特异性引物,通过优化PCR扩增条件和体系,建立了可检测PBoV G1的PCR方法。针对该检测方法进行特异性和敏感性的验证。结果显示,该方法仅对PBoV G1具有特异性,可扩增出特异性目的条带,对其他猪常见肠道病毒均无交叉反应;PBoV G1标准阳性质粒最低检出量为5.46×10^(3)拷贝/μL。应用建立的检测方法对463份猪腹泻样品进行检测,结果显示PBoV G1的阳性率为25.05%(116/463)。建立的PCR方法特异性强、灵敏度高,对猪群中PBoV G1的病原检测、疫情防控及流行病学调查等具有重要意义。
To establish a PCR assay for rapid detection of Porcine Bocavirus Genogroup G1(PBoV G1),a pair of specific primers were designed based on the conserved sequence of VP1 gene from the newly available PBoV G1 strains in GenBank.By optimizing the PCR amplification conditions and system,a PCR assay for the detection of PBoV G1 was established.The specificity and sensitivity of this detection method were verified,and preliminary clinical application was carried out.The results of the specificity test show that this assay is exclusively specific to PBoV G1,amplifying a distinct target band,while showing no cross-reactivity with other common porcine enteric viruses.The results of the sensitivity test show that the limit of detection for the PBoV G1 standard positive plasmid is 5.46×10^(3) copies/μL.Among 463 porcine diarrhea samples tested by the developed assay,PBoV G1 is detected in 116 cases,yielding a positive rate of 25.05%.It indicates that the PCR assay established by this research institute has strong specificity and high sensitivity,and is of great significance for the pathogen detection,epidemic prevention and control,and epidemiological investigation of PBoV G1 in pigs.
作者
温悦
张卓威
高定焯
赵宇琛
张亚
刘琪
刘昱涵
付朋飞
WEN Yue;ZHANG Zhuo-wei;GAO Ding-zhuo;ZHAO Yu-chen;ZHANG Ya;LIU Qi;LIU Yu-han;FU Peng-fei(School of Sciences and Engineering,Henan University of Urban Construction,Pingdingshan 467036,China)
出处
《河南城建学院学报》
2025年第2期114-120,共7页
Journal of Henan University of Urban Construction
基金
河南城建学院2024年大学生创新创业训练计划资助项目(202411765035)。
作者简介
通信作者:付朋飞(1985—),男,河南平顶山人,博士,讲师,研究方向为分子病毒学和分子免疫学。E-mail:fpfwdm@126.com。
