不同检测材料对ELISA检测禽白血病病毒-p27抗原结果的影响  

Effect of Different Test Materials on the Detected Results of Avian Leukosis Virus-p27 Antigen by ELISA

作　　者：沈学怀[1] 张丹俊[1] 潘孝成[1] 赵瑞宏[1] 胡晓苗[1] 戴银[1] 侯红艳[1] 朱传民[1] 

机构地区：[1]安徽省农业科学院畜牧兽医研究所,合肥230031

出　　处：《中国畜牧兽医》2015年第5期1294-1300,共7页China Animal Husbandry & Veterinary Medicine

基　　金：国家现代农业产业技术体系项目(CARS-41);安徽省家禽产业技术体系专项;安徽省农科院科技创新团队项目(11C0404);国家自然科学基金"MHCⅠ分子靶向性疫苗载体的构建及其免疫作用的机理"(31302044)

摘　　要：为探讨不同检测材料对ELISA检测禽白血病病毒(ALV)-p27抗原结果的影响,试验首先对180日龄地方品种母鸡的泄殖腔棉拭子ALV-p27抗原进行ELISA检测,选取部分阳性鸡和阴性鸡进行分组试验,采用ELISA对试验鸡血清、蛋清和病毒分离的细胞培养液进行ALV-p27抗原检测,采用RT-PCR方法检测血液ALV特异性基因。结果显示,阳性组中以血清、蛋清和细胞培养液作为ELISA ALV-p27抗原检测材料,其检测阳性率均低于泄殖腔棉拭子,血清检测的阳性个体包含全部蛋清和细胞培养液ELISA检测的阳性个体。ELISA检测数据的相关性分析显示,只有血清和细胞培养液检测数据间存在显著性相关,线性关系方程为Y(细胞上清)=1.8439X(血清)-0.1469,R2=0.937;阳性组中ALV-p27基因检测阳性率低于泄殖腔棉拭子,但高于血清、蛋清和细胞培养液,其包含所有血清ELISA检测的阳性样品;外源性ALV-J gp85基因阳性率仅为29.17%,且阳性样品均属于血清ELISA阳性样品。综上所述,成年鸡以泄殖腔棉拭子作为ELISA ALV-p27抗原检测材料存在假阳性结果,蛋清和细胞培养液作为检测材料存在漏检的可能,血清作为ELISA检测材能够更准确地反映成年鸡群ALV感染状态。In order to study the effect of different test materials on the detected results of avian leukosis virus （ALV）-p27 antigen by ELISA,the cloacal swabs ALV-p27 antigen were detected by ELISA in 180-day local breeds hen,and the parts of the positive and negative chicken was se- lected to group test, the serum, egg whites and cell supernatant ALV-p27 antigen were detected by ELISA, the specific genes of ALV were detected by RT-PCR in blood. The results showed that serum, egg white and cell supernatant as ELISA test materials, the positive rate lower of than clo- acal swabs,and serum ALV-p27 positive samples include all egg whites and cell supernatant posi- tive samples in positive group. It was a significant correlation between ELISAs with serum and cell supernatant （linear equation：Y= 1. 8439X--0. 1469 ,the correl was 0. 937）. In positive group, ALV-p27 gene positive rate lower than cloacal swabs ELISA,but higher than the serum,egg and cell culture medium, and ALV-p27 gene positive samples include serum positive samples byELISA. ALV-J gp85 gene positive rate of 29.17% ,and all positive samples were included in the serum ALV-p27 positive samples. The results suggested that the cloacal swabs as test material may occur false positive results, and egg whites and cell supernatant may occur undetected by ELISA ALV-p27 antigen assay in adult chicken, serum as test material in ELISA could more accurately reflect the state of adult chickens infected with ALV.

关 键 词：禽白血病病毒 ALV-p27抗原 ELISA 检测材料 

分 类 号：S858.31[农业科学—临床兽医学]