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3种猫疱疹病毒Ⅰ型检测方法的比较
萧晟,熊冉,苑文涛,王广操,徐敏,陈文华
0
(芜湖职业技术学院生物工程学院,芜湖241003;南京灵越检测技术有限公司,南京 210000)
摘要:
比较传统PCR、荧光定量PCR以及胶体金速测卡对于猫疱疹病毒Ⅰ型(FHV-1)检测的敏感性及准确性。设计引物,优化反应条件并扩增目的片段,定向插入pMD-19T克隆载体,最后用建立的2种PCR检测方法及猫疱疹病毒1型快速检测卡对21份临床疑似猫疱疹病毒感染的分泌物进行检测。荧光定量PCR和传统PCR检测的最低拷贝数分别为20拷贝和207拷贝;荧光定量PCR、传统PCR以及胶体金快速检测卡的检出率分别为71%、57%和29%。传统PCR及荧光定量PCR检测方法均具有良好的特异性,其中荧光定量PCR特异性显著,相比猫疱疹病毒1型快速检测卡敏感性好,准确率更高。
关键词:  猫疱疹病毒1型  传统PCR  荧光定量PCR  特异性  敏感性
DOI:10.13610/j.cnki.1672-352x.20190716.004
基金项目:安徽省高校学科(专业)拔尖人才学术资助重点项目(gxbjZD2016120), 安徽省特色(品牌)专业(2016tszy087)、安徽省高校科学研究项目(KJ2018A0693)共同资助。
Comparison of detection on feline herpesvirus-1 by three methods
XIAO Sheng,XIONG Ran,YUAN Wentao,WANG Guangcao,XU Min,CHEN Wenhua
(Wuhu Institute of Technology, Wuhu 241003;Lingyue?Technology?Laboratory?Co.,Ltd., Nanjing 210000)
Abstract:
This study compared the sensitivity and accuracy of detection on feline herpesvirus-1 by using three methods of traditional PCR, Real-time?PCR and colloidal gold detection card. Two methods of traditional ?PCR and Real-time PCR based on Taq Man probe technology were established to detect feline herpesvirus-1 in?our lab. The minimum of copies detected by fluorogenic quantitative PCR and traditional PCR were 20 and 207, respectively. The two well?established?methods?in?our?lab?and?the?rapid?detection?card?of Feline herpesvirus-1 were used to detect 21 secretion?suspected?Feline?herpesvirus?infection?clinically. It?showed that the detection rates of Real- time PC, traditional?PCR?and?quick?test?card?were?71%,57%?and?29%, respectively. The result indicated that compared with the rapid detection card of cat herpesvirus type 1, the detection method of traditional PCR and Real- time PCR is?more?sensitive?and?accurate, especially?Real-time?PCR which can provide more accurate ?results for clinical diagnosis.
Key words:  FHV-1  traditional PCR  fluorogenic quantitative PCR  accuracy  sensitivity

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