犬细环病毒SYBR GreenⅠ荧光定量PCR检测方法的建立
发布时间：2018-12-13 10:58
【摘要】：为建立犬细环病毒(TTCV)的快速检测方法,本研究根据GenBank中登录的TTCV ORF7基因设计合成一对特异性引物,并对反应条件和反应体系进行优化,建立了检测TTCV的SYBR Green I荧光定量PCR方法。结果显示:建立的荧光定量PCR方法 Ct值与标准品模板在5.82×10~9拷贝/μL~5.82×10~3拷贝/μL范围内呈良好的线性关系。该检测方法仅对TTCV检测为阳性,而对狂犬病病毒、犬冠状病毒、犬细小病毒、犬瘟热病毒和犬副流感病毒均无特异扩增;该检测方法灵敏度可达5.82×103拷贝/μL。对27份犬血清样品检测结果显示,建立的荧光定量PCR阳性检测率为11.11%,而普通PCR方法检测阳性率为3.7%。本实验建立了TTCV SYBR Green Ⅰ荧光定量PCR检测方法,对TTCV诊断及致病机制的研究提供了高通量的定量检测方法。
[Abstract]:In order to establish a rapid detection method for (TTCV) of canine fine loop virus, a pair of specific primers were designed and synthesized according to the TTCV ORF7 gene registered in GenBank, and the reaction conditions and reaction system were optimized. A SYBR Green I fluorescence quantitative PCR method for the detection of TTCV was established. The results showed that there was a good linear relationship between the Ct value and the standard sample template in the range of 5.82 脳 10 ~ (9) copies / 渭 L ~ 5.82 脳 10 ~ (-3) copies / 渭 L. The sensitivity of this method was 5.82 脳 10 ~ 3 copies / 渭 L for rabies virus, canine coronavirus, canine parvovirus, canine distemper virus and canine parainfluenza virus. The positive rate of fluorescent quantitative PCR was 11.11% in 27 canine serum samples, while the positive rate of normal PCR method was 3.7%. In this study, a fluorescence quantitative PCR assay for the detection of TTCV SYBR Green 鈪，
本文编号：2376426
[Abstract]:In order to establish a rapid detection method for (TTCV) of canine fine loop virus, a pair of specific primers were designed and synthesized according to the TTCV ORF7 gene registered in GenBank, and the reaction conditions and reaction system were optimized. A SYBR Green I fluorescence quantitative PCR method for the detection of TTCV was established. The results showed that there was a good linear relationship between the Ct value and the standard sample template in the range of 5.82 脳 10 ~ (9) copies / 渭 L ~ 5.82 脳 10 ~ (-3) copies / 渭 L. The sensitivity of this method was 5.82 脳 10 ~ 3 copies / 渭 L for rabies virus, canine coronavirus, canine parvovirus, canine distemper virus and canine parainfluenza virus. The positive rate of fluorescent quantitative PCR was 11.11% in 27 canine serum samples, while the positive rate of normal PCR method was 3.7%. In this study, a fluorescence quantitative PCR assay for the detection of TTCV SYBR Green 鈪，
本文编号：2376426
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