摘要
本研究建立了检测狂犬病病毒抗原的夹心间接斑点酶联免疫吸附试验(SI-Dot-ELISA).本法检出狂犬病病毒抗原的最低浓度为:0.01IU/mL的标准抗原,1:100000 (相当于20 LD_(50))的狂犬病病毒CVS株和鹿8202株的鼠脑悬液,1:400(相当于7906TCID_(50)/mL)的狂犬病病毒SAG株的细胞培养物.对多种健康动物的组织、健康细胞培养物以及犬瘟热病毒、犬腺病毒等检测均为阴性.用SI—Dot—ELISA、夹心间接ELISA(SI—ELISA)和小鼠脑内接种3种方法,检测了388份材料,检出的阳性份数分别为173、171和179,经统计学处理,3种方法在狂犬病病毒抗原的检测上,可以相互代替.甲醛灭活病毒不影响本方法的检测,而加入氢氧化铝胶后的病毒悬液则不能用本法检测.本法适用于狂犬病的诊断、流行病学调查、疫苗效价的测定和实验研究中病毒抗原的测定.
Sandwich indirect Dot-ELISA (SI-Dot-ELISA) was developed to detect the rabies virus antigen. The standard rabies virus antigen (0. 01 IU/mL), 1 : 100000 (about 20 LD50) of the mouse brain antigen of CVS rabies virus strain and 8202 strain of deer rabies virus might be successfully detected. Normal tissue samples from some kinds of animals, the normal cell culture and some other viruses did not reveal the reactions. 388 samples were tested by SI-Dot-ELISA, SI-ELISA and mouse intracerebral inoculation, 173,171 and 179 positive results were obtained respectively. The results were analysed statistically and it was shown that they could be substituted each other. The test was not affected by the inactivation process of the virus preparation with formalin, but it failed to perform when aluminium hydroxide was added into the vitus suspension. The method is suitable for diagnosis and epidemiological surveys of rabies and determination of the efficacy of rabies vaccine and antigen titers of rabies virus.
