表达鸡毒支原体TM1蛋白基因重组新城疫病毒的构建及其免疫原性研究  

Construction of recombinant Newcastle disease virus expressing TM1 gene of Mycoplasma gallisepticum and its immune response in chickens

作　　者：冯秋霖[1] 刘茂军[2] 祁芳[1] 陈曦[1] 邵国青[2] 葛金英[1] 步志高[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部兽医公共卫生重点开放实验室,黑龙江哈尔滨150001 [2]江苏省农业科学院兽医研究所,江苏南京210095

出　　处：《中国预防兽医学报》2013年第10期779-782,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30800816)

摘　　要：为研制安全、有效的新型鸡毒支原体(MG)疫苗,本研究利用新城疫病毒(NDV)LaSota弱毒疫苗株反向遗传操作系统,构建出表达MG TM1蛋白的重组病毒rLa-TM1,并对其生物学特性进行鉴定,评估其作为MG活载体疫苗的安全性和有效性。通过免疫荧光试验表明TM1蛋白获得正确表达;经测定,重组病毒平均鸡胚致死时间(MDT)为132 h,脑内致死指数(ICPI)和静脉内致病指数(IVPI)均为0,保持了LaSota亲本株的低致病性;rLa-TM1接种鸡胚所收集尿囊液中的病毒滴度可高达每毫升109.5EID50以上,证明重组病毒保持了LaSota亲本株高滴度鸡胚生长特性;重组病毒rLa-TM1接种1周龄的雏鸡后,可以诱导显著的MG抗体反应;攻毒保护试验证明其保护率可以达90%,而对照组全部发病(10/10)。本研究表明重组病毒rLa-TM1可以作为MG重组病毒活载体疫苗的候选疫苗。In order to develop a safe and efficient recombinant Mycoplasma gallisepticum （MG） vaccine based on Newcastle disease virus （NDV） LaSota vaccine strain as viral vector, we constructed and rescued a recombinant NDV （rLa-TM1） which expressed TM1 protein of MG by reverse genetic techniques. The expression of MG TM1 protein from recombinant virus was confirmed in the infected BHK cells by indirect immunofluorescence assay. Pathogenic test shown that the rLa-TMI retained NDV high-titer growth property and low pathogenicity in embryonated chicken eggs. Moreover, the rLa-TM1 induced high titer of antibody against MG in one week post vaccination in chickens which provided up to 90% of protection against MG challenge. These results demonstrated that the rLa-TM1 had the potential of being used as a novel MG vaccine.

关 键 词：鸡毒支原体 反向遗传操作系统 重组新城疫病毒 

分 类 号：S852.62[农业科学—基础兽医学]