摘要
为探讨肺组织中猪生殖与呼吸综合征病毒(PRRSV)抗原的定位诊断方法,制备了兔抗PRRSV抗体,采用免疫组织化学Envision法对疑似PRRS病猪肺组织中的病毒抗原进行了定位和半定量检测。SDS-PAGE和Western-blotting检测结果表明,制得的兔抗PRRSV IgG纯度较高,具有良好的特异性,可用于原位检测病猪肺组织中PRRSV抗原的分布;Envision法的检测结果显示,PRRSV抗原的阳性表达产物主要出现在肺巨噬细胞胞浆内,其次为肺泡上皮细胞和细支气管黏膜上皮细胞;通过对5份经RT-PCR检测确认的PRRS病猪肺组织进行检测,均有较高的阳性细胞表达,平均阳性细胞率为46.5%。证明Envi-sion法具有高敏感、低背景、快速简便的特点,可用于肺组织中PRRSV抗原的快速检测。
To establish a method for locating diagnosis of porcine reproductive and respiratory syn- drome virus（PRRSV） antigen in lung tissues,rabbit IgGs against PRRSV were prepared and used as primary antibodies in the method of immunohistochemistry for detection of PRRSV antigens in the infected lung tissues from pigs. The antibodies were purified and the purity and specificity were identified by SDS-PAGE and Western-blot,respectively. Lung tissues from five pigs with PRRS were tested with the estab-lished method, which clearly demonstrated the abundant presence of viral antigens in the cytoplasm of alveolar macrophages,and to less extent in alveolar and bronchiole epithelial cells. The results showed that the Envision immunohistochemistry was a rapid and convenient method for detection of PRRSV with high sensitivity and specificity,and would be used for the rapid detection of PRRSV antigen in the lung of swine.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2009年第3期237-240,共4页
Chinese Veterinary Science
基金
广东省佛山市科技发展专项资金项目(2005020022)
国家“十一五”科技支撑计划项目(2006BAD06A12)
