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6种猪病毒病微流控芯片多重检测方法的建立

梅力 冯小宇 王培 刘海莹 李志军 张启龙 高晓龙 陈会玲 赵建晴 马银平

中国动物检疫2025,Vol.42Issue(5):111-119,9.
中国动物检疫2025,Vol.42Issue(5):111-119,9.DOI:10.3969/j.issn.1005-944X.2025.05.018

6种猪病毒病微流控芯片多重检测方法的建立

Establishment of a Microfluidic Chip Method for Multiple Detection of 6 Porcine Viral Diseases

梅力 1冯小宇 1王培 1刘海莹 1李志军 1张启龙 1高晓龙 1陈会玲 1赵建晴 2马银平2

作者信息

  • 1. 北京市动物疫病预防控制中心,北京 102629
  • 2. 北京博奥晶典生物技术有限公司,北京 101111
  • 折叠

摘要

Abstract

In order to simultaneously detect the classical strain of porcine reproductive and respiratory syndrome virus(PRRSV),PRRSV highly pathogenic strain(HP-PRRSV),classical swine fever virus(CSFV),pseudorabies virus(PRV),porcine circovirus type 2(PCV2)and porcine parvovirus(PPV),primers were designed according to the genetically conserved regions of the 6 viruses,respectively,and embedded in reaction pools of isothermal amplification microfluidic chip,a microfluidic chip method was established to simultaneously detect all the above viruses through optimizing the concentration of primers,reaction temperature and time.The results showed that the lowest detection limits of the established method for the plasmid templates of the 6 viruses were all 103 copies/µL;specificity analysis showed that viruses or samples presented an"S"-shaped amplification curve only in the reaction pool corresponding to their specific detection indicators;good reproducibility was observed,the intra-and inter-batch coefficients of variation were less than 5%;for 40 clinical samples,the compliance rate of the method with commercial quantitative fluorescence detection kits reached above 95%.In conclusion,a microfluidic chip method was successfully established for 6 porcine viral diseases,which presented high sensitivity,strong specificity and good reproducibility,achieving joint detection for multiple animal diseases.

关键词

猪繁殖与呼吸综合征病毒/猪瘟病毒/猪伪狂犬病病毒/猪圆环病毒2型/猪细小病毒/微流控芯片

Key words

PRRSV/CSFV/PRV/PCV2/PPV/microfluidic chip

分类

农业科技

引用本文复制引用

梅力,冯小宇,王培,刘海莹,李志军,张启龙,高晓龙,陈会玲,赵建晴,马银平..6种猪病毒病微流控芯片多重检测方法的建立[J].中国动物检疫,2025,42(5):111-119,9.

基金项目

北京市动物疫病预防控制中心综合保障项目(11000024Y000002862075) (11000024Y000002862075)

中国动物检疫

1005-944X

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