›› 2015, Vol. 42 ›› Issue (5): 1294-1300.doi: 10.16431/j.cnki.1671-7236.2015.05.039

• 疾病防治 • 上一篇    下一篇

不同检测材料对ELISA检测禽白血病病毒-p27抗原结果的影响

沈学怀, 张丹俊, 潘孝成, 赵瑞宏, 胡晓苗, 戴银, 侯红艳, 朱传民   

  1. 安徽省农业科学院畜牧兽医研究所, 合肥 230031
  • 收稿日期:2014-09-10 出版日期:2015-05-20 发布日期:2015-06-02
  • 通讯作者: 张丹俊 E-mail:zhangdj694@sina.com
  • 作者简介:沈学怀(1986-),男,安徽凤阳人,硕士,研究方向:畜禽传染病,E-mail:xuehuaishen1986@163.com
  • 基金资助:
    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)

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

SHEN Xue-huai, ZHANG Dan-jun, PAN Xiao-cheng, ZHAO Rui-hong, HU Xiao-miao, DAI Yin, HOU Hong-yan, ZHU Chuan-min   

  1. Institute of Animal and Veterinary Science, Anhui Academy of Agricultural Sciences, Hefei 230031, China
  • Received:2014-09-10 Online:2015-05-20 Published:2015-06-02

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。

关键词: 禽白血病病毒; ALV-p27抗原; ELISA; 检测材料

Abstract: 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 selected 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 cloacal swabs, and serum ALV-p27 positive samples include all egg whites and cell supernatant positive 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 by ELISA.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.

Key words: avian leukosis virus; ALV-p27 antigen; ELISA; test materials

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