RDS-ELISA的建立及其在检测JEV抗原和抗体中的应用

Establishment of RDS-ELISA and It's a Pplication in the Detection of Antigens and Antibodies of JEV

通过用鞣酸处理微板、在稀释液中加入4％PEG、提高NaC1浓度,用鸡蛋清代替鸡血清、TMBS代替OPD底物、自来水代替PBS洗液、提高试验温度、缩短反应时间等措施,建立了一种快速酶联免疫吸附试验双夹心法(RDS-ELISA),并用该法对日本脑炎病毒(JEV)及其单抗进行了测定。不仅整个试验可在1小时内完成,而且所得结果与常规双夹心法相同,比血凝试验约高4倍;抗体效价与常规法相同或高10倍,比间接法高10～10,000倍。该法是一种快速、简便、特异、敏感的方法。

By Protreating the microplate with tannie acid, adding 4% PEG to the dilution fluid to increase NaCl concentration in it, substituting egg white for chicken serum, TMBS for OPD, normal saline solution for PBS as the washing solution, raising reaction temperature shorting reaction time, we have established a rapid double sandwish ELISA, and applied this method in the detection of Japanese B - encephalitis virus antigens and McAbs. Beginning from the microplate coating, the whole test could be finished within one hour, wfth antigenic titre being four times higher than that of hemagglutination test, or as high as that of conventional double sandwish ELISA, and McAb titre ten to ten thousend times higher than that of indirect ELISA, or as high as ten times higher than that of conventional double sandwish ELISA. It showed that this method was rapid, simple, specific and sensitive.