鸡细小病毒荧光LAMP检测方法的建立

本试验旨在建立一种可鉴别鸡细小病毒(ChPV)的环介导等温扩增(LAMP)检测方法。根据鸡细小病毒NS1基因的保守序列,设计了1套特异性引物,在该套引物的F3与B3区域中设计1个探针,其探针用FAM标记,优化建立了能鉴别ChPV的荧光LAMP检测方法,并使用所建立的方法对21份临床样品进行检测。结果显示,该方法特异性好,能检测到ChPV,并与其他禽类病毒无交叉反应;灵敏度高,最低能够检测1.02×10^-5ng/μL的ChPV DNA模板;对临床可疑样品的检出率为24%。表明,本研究建立的ChPV荧光LAMP方法具有简便快速、特异性好、敏感性高等优点,可用于检测ChPV。

Establishment of fluorescence LAMP for detection of chicken parvovirus

The purpose of this study was to establish a loop-mediated isothermal amplification(LAMP) method for the identification of chicken parvovirus(ChPV).According to the conserved sequence of ChPV NS1 gene, a set of specific primers was designed. And a probe was designed in the F3 and B3 regions of this set of primers.The probe was labelled with FAM to optimize the establishment of fluorescent LAMP detection method for ChPV identification.The established method was used to detect 21 clinical samples.The results showed that the detection method was specific,could detect ChPV,had no cross-reaction with other avian pathogens.The method had high sensitivity and capable of detecting a minimum of 1.02×10^-5 ng/μl-1 ChPV virus DNA template.The detection rate of clinical suspicious samples was 24%.The fluorescence LAMP method of chicken parvovirus established in this study has the advantages of simplicity,rapidity,specificity and high sensitivity.It can effectively prevent the epidemic caused by ChPV.