鸽疱疹病毒gB基因SYBR GreenⅠ荧光定量PCR检测方法的建立与应用  

Establishment and Application of SYBR GreenⅠFluorescent Quantitative PCR Based on gB Gene for Detection of Pigeon Herpesvirus

作　　者：李鹏 苏和 韩慧敏 孟海 王昊 王凤雪 温永俊[1] LI Peng;SU He;HAN Huimin;MENG Hai;WANG Hao;WANG Fengxue;WEN Yongjun(Key Laboratory for Clinical Diagnosis and Treatment of Animal Diseases of Ministry of Agriculture and Rural Affairs,College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot,Inner Mongolia 010018;Animal Disease Prevention and Control Center of Hohhot City,Hohhot,Inner Mongolia 010018)

机构地区：[1]内蒙古农业大学兽医学院,农业农村部动物疾病临床诊断与治疗重点实验室,内蒙古呼和浩特010018 [2]呼和浩特市动物疫病预防控制中心,内蒙古呼和浩特010018

出　　处：《中国家禽》2025年第3期93-99,共7页China Poultry

基　　金：国家自然科学基金地区项目(32260894);内蒙古自治区教育厅“高校青年科技英才”项目(NJYT22043)。

摘　　要：为建立检测鸽疱疹病毒(PiHV)的SYBR GreenⅠ荧光定量PCR(qPCR)方法,研究根据PiHV gB基因的高度保守区域设计特异性引物,构建质粒作为标准品建立PiHV qPCR检测方法,并进行敏感性和特异性试验。结果显示:建立的PiHV qPCR方法最低DNA检出浓度为8.62×10^(2)拷贝/μL,敏感性比普通PCR方法高100倍,可特异性区分鸽疱疹病毒与其他多种常见感染鸽的细菌和病毒,其批内变异系数小于1%,批间变异系数小于2%;应用该方法对呼和浩特市及周边地区赛鸽公棚和鸽养殖场采集的30份疑似病鸽肝脏样品进行检测,可快速准确地检出样品中的PiHV,并且检出率比普通PCR方法高。研究表明,建立的PiHV qPCR检测方法特异性强,敏感性较高,能够应用于赛鸽、肉鸽等的鸽疱疹病毒快速检测。In order to establish a SYBR GreenⅠfluorescent quantitative PCR(qPCR)method for pigeon herpesvirus(PiHV),the study was designed specific primers according to the highly conserved region of gB gene of PiHV,and the plasmids were constructed as standard materials to establishh the SYBR GreenⅠfluorescent quantitative polymerase chain reaction(qPCR)method for PiHV,and the sensitivity and specificity of the method were analyzed.The results showed that the minimum DNA detection limit of the established PiHV qPCR method was 8.62×10^(2) copies/μL,and the sensitivity was 100 times higher than that of the common PCR method.At the same time,the method could specifically distinguish pigeon herpesvirus and other common pigeon susceptible pathogens,and the intra-batch coefficient of variation was less than 1%,and inter-batch coefficient of variation was less than 2%.The established method could quickly and accurately detect PiHV in 30 liver samples of suspected diseased pigeons collected from racing pigeon stables and pigeon farms in Hohhot and surrounding areas,the detection rate of which was higher than that of the common PCR method.The results indicated that the established PiHV qPCR method had strong specificity and high sensitivity,and could be applied to the rapid detection of PiHV in racing pigeons,meat pigeons and other groups.

关 键 词：鸽疱疹病毒 SYBR Green 荧光定量PCR 聚合酶链式反应 GB基因 

分 类 号：S855.3[农业科学—临床兽医学]