SDS-PAGE法检测产气荚膜梭菌毒素型研究

Research on contrast of SDS-PAGE identifying toxin types of Clostridium perfringens

为了探索一种快速、准确检测产气荚膜梭菌毒素型的方法，以便有效预防控制该病的流行和发生，利用SDS-PAGE方法，对通过EusA鉴定的20株（A型16株、C型3株和D型1株）德国牛舍环境气源性产气荚膜梭菌分离株，以及通过Multiplex-PCR鉴定的57株山东省疫区产气荚膜梭菌分离株外毒素蛋白的相对分子质量进行测定，以确定毒素的种类，进而确定菌株的毒素型。结果表明，产气荚膜梭菌标准株、德国牛舍环境气源性产气荚膜梭菌分离株、山东省疫区产气荚膜梭菌分离株粗提外毒素的质量浓度分别为3．261，2．238～3．333，1．451～3．109mg／mL，精提外毒素的质量浓度分别为0．803，0．521～O．999，0．530～0．812mg／mL。SDS-PAGE法检测的20株德国牛舍环境气源性产气荚膜梭菌分离株中A型17株、C型2株、D型1株，与EuSA检测结果的符合率为95．0％；SDS-PAGE检测的57株山东省疫区产气荚膜梭菌分离株全部是A型，与Multiplex-PCR检测结果的符合率为100％。说明应用SDS-PAGE方法对产气荚膜梭菌的检测结果与ELISA方法的检测结果有一定的差异性，与Multi-plex-PCR方法的检测结果一致。

This experiment was carried out by using SDS-PAGE method to determine the molecular weight of exotoxin protein produced by 20 （16 of type A,3 of type C and 1 of type D） airborne Clostridium perfringens strains from cow stable that the toxin types were examined by ELISA and 57 Clostridium perfringens strains selected from Shandong Province had been identified by multi-PCR to confirm the kind of toxin,then their toxin types were obtained. The detecting results showed that the quality concentrations of exotoxin by coarse distill of Clostridiurn perfringens standard strains, airborne Clostridiurn perfringens samples from cow stable in German and Clostridium perfringens strains separated from Shandong Prov- ince were 3. 261,2. 238-3. 333 and 1. 451-3. 109 mg/mL,respectively;the quality concentrations of exo- toxin by extractive distill were 0. 803,0. 521-0. 999 and 0. 530-0. 812 mg/mL,respectively. The detecting results of 20 airborne Clostridiurn perfringens strains from cow stable were 17 of type A,2 of type C and 1 of type D,and the accordance with ELISA was 95.0~;the detecting results of 57 Clostridiurn perfrin- gens strains separated from Shandong Province were all of type A,and the accordance with multi-PCR was 100%. The results showed that the difference existed between ELISA method and SDS-PAGE method andthere was coherence between multi-PCR method and SDS-PAGE method.