犬博卡病毒PCR检测方法的建立及其应用

Establishment and Application of a PCR Detection Method for Canine Bocavirus

[目的]建立犬博卡病毒(Canine bocavirus,CBo V)的PCR检测方法,了解其流行情况。[方法]根据Gen Bank数据库上CBo V的VP2基因序列合成1对引物,建立PCR检测方法,并运用该方法对临床样品进行检测。[结果]特异性试验中,除CBo V有目的条带出现外,犬细小病毒(CPV)、犬瘟热病毒(CDV)、犬传染性肝炎(ICHV)、犬副流感病毒(CPIV)、猫瘟热(FDV)等均无条带出现。敏感性试验表明,此检测方法对CBo V的最低检测量为4.806 2 pg/μL。重复性试验表明,多次重复试验结果一致,表明此方法重复性好。425份样品中仅有1份样品扩增出目的条带,经测序比对鉴定为CBo V。初步的流行病学调查结果表明,CBo V在犬中的流行率与感染率极低。[结论]建立的PCR检测方法具有特异性强、敏感性高、可重复性强等特点。

[Objective] The aim was to establish canine bocavirus PCR detection method and study its prevalence.[Method] A pair of primers were synthesized according to the sequence of VP2 gene of CBo V on Genbank database,and the PCR detection method was established and was applied for detection of clinical sample.[Result]In specific tests,except for CBo V with the emergence of the band,CPV,CDV,ICHV,CPIV,FDV and so on were not brought to appear. Sensitivity experiments showed that the minimum detection amount of CBo V was 4. 806 2 pg / μL. The results of repeated experiments were consistent,indicating that the method was reproducible. Among 425 samples,1 sample was amplified with the target bands,and the result was determined as CBo V. Preliminary epidemiological survey showed that the prevalence rate and infection rate of CBo V in dogs was very low.[Conclusion] The established detection method has characteristics of strong strong specificity,high sensitivity and strong repeatability.