核酸斑点杂交法与ELISA在SPF鸡检测弱毒疫苗中ALV污染的应用与比较  

Application and Comparison of Nucleic Acid Spot Hybridization and ELISA Detecting ALV within the Blood of SPF Chickens via Injection of Contaminated Attenuated Vaccine

作　　者：栾怀彪 赵颖洁[1] 吕艳艳[1] 徐颖 崔治中[1] 常爽[1] 赵鹏[1] 

机构地区：[1]山东农业大学动物医学院,山东泰安271018 [2]青岛出入境检验检疫局,山东青岛266001

出　　处：《中国家禽》2016年第4期10-13,共4页China Poultry

基　　金：公益性行业(农业)科研专项经费项目(201203055)

摘　　要：本研究对SPF鸡检查法在检测弱毒疫苗中低剂量禽白血病病毒(ALV)污染时的核酸斑点杂交法和ELISA进行了比较。分别以10 TCID_(50)/羽份和5 TCID50/羽份ALV-A人为污染一批商品化新城疫(ND)活疫苗,将污染ND疫苗各接种10只SPF鸡,以ELISA定期检测ALV抗体并利用PCR结合核酸斑点杂交法检测血液中ALV核酸。结果显示,在免疫污染疫苗后连续3周内ALV抗体全部为阴性,而核酸斑点杂交法检测表明自免疫后两周开始就出现ALV阳性。结果提示在利用经典的SPF鸡检查法检测弱毒疫苗中ALV污染时,结合对病原核酸的检测有助于节省检测时间和降低漏检率。In this study,nucleic acid spot hybridization（NASH）and ELISA method were used to detect low-dose avian leukosis virus（ALV）in SPF chickens inoculated contaminated attenuated vaccines. The quantified ALV-A with10 TCID_（50） and 5 TCID_（50） was diluted respectively in a batch of commercial attenuated Newcastle disease（ND）vaccine.The contaminated vaccine with two different doses of ALV-A was inoculated to 10 SPF chickens respectively. Antibody against ALV-A and nucleic acid of ALV-A were regularly detected by ELISA and NASH. The results showed that all antibody against ALV were negative after inoculation of the contaminated vaccine for three consecutive weeks,but positive after two weeks with NASH,which suggested that when detecting low-dose ALV in SPF chickens inoculated contaminated attenuated vaccine,the testing time was saved and misdetection rate was reduced through combined with NASH.

关 键 词：禽白血病病毒 弱毒疫苗 污染 SPF鸡 

分 类 号：S855.3[农业科学—临床兽医学]