CAJ | 학술논문

以灭活的狂犬病病毒CVS株细胞毒免疫BALB/c小鼠，通过间接ELISA法和Western-blot筛选获得针对磷蛋白的单克隆抗体4株：1C9、4B10、2G12、4G5，其中1C9针对氨基端保守表位。以亲和层析法纯化1C9单抗腹水，异硫氰酸荧光黄标记制备荧光抗体。以1C9磷蛋白荧光抗体与本实验室研制的狂犬病核蛋白免疫荧光抗原检测试剂盒，对本实验室收集的501份疑似狂犬病鼬獾、蝙蝠、犬和黄鼬的脑组织样品进行直接免疫荧光平行检测。结果显示，两种检测手段对基因1型狂犬病毒的检出结果完全一致，而蝙蝠源Irkut病毒仅能以磷蛋白单抗1C9检出。本研究成功获得了与我国现有不同基因型狂犬病毒良好反应的抗狂犬病磷蛋白单抗，并应用于狂犬病的直接免疫荧光检测，为狂犬病诊断提供了敏感性和可靠性良好的诊断试剂。
이멸활적광견병병독CVS주세포독면역BALB/c소서，통과간접ELISA법화Western-blot사선획득침대린단백적단극륭항체4주：1C9、4B10、2G12、4G5，기중1C9침대안기단보수표위。이친화층석법순화1C9단항복수，이류청산형광황표기제비형광항체。이1C9린단백형광항체여본실험실연제적광견병핵단백면역형광항원검측시제합，대본실험실수집적501빈의사광견병유환、편복、견화황유적뇌조직양품진행직접면역형광평행검측。결과현시，량충검측수단대기인1형광견병독적검출결과완전일치，이편복원Irkut병독부능이린단백단항1C9검출。본연구성공획득료여아국현유불동기인형광견병독량호반응적항광견병린단백단항，병응용우광견병적직접면역형광검측，위광견병진단제공료민감성화가고성량호적진단시제。
Four monoclonal antibodies against Rabies virus phosphoprotein were produced by immunization with Rabies virus strain CVS in BALB/c mice. After fusion, antibodies in the supernatant of the hybrid cells were respectively detected by ELISA test and western-blot method. Four hybrid cells secreting antibody binding to rabies virus , i.e. 1C9, 4B10, 2G12 and4G5 strains. The 1C9 monoclonal lgG antibody was purified by affinity chromatography method and then labeled with FITC. 501 brain samples collected from ferret badgers, bats, dogs and weasels were detected using the labeled 1C9 antibody and FITC-labeled rabies virus N protein-specific monoclonal antibody (made in our laboratory). The results showed that the labeled 1C9 antibody can detect rabies-related lyssaviruses isolated in China including Rabies virus and Irkut virus, however, the N protein-specific monoclonal antibody only can detect Rabies virus. It suggests that the labeled 1C9 antibody can be used as a rabies diagnostic reagent in China.