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目的研究和比较中国艰难梭菌A-B+高分离株BJ08和欧洲流行A+B+高毒株(BI/NAP1/027)UK1的tcdA、tcdB转录和毒素表达。方法每隔3h提取艰难梭菌和培养上清,用实时荧光定量PCR的方法测定各个时间段BJ08和UK1菌株tcdA、tcdB基因的表达;用ELISA的方法检测BJ08和UK1菌株各个时间段的细胞和培养上清液中A毒素、B毒素的含量。结果 BJ08的生长速率显著大于UK1;BJ08不表达A毒素,但是检测到tcdA基因的转录,tcdA基因表达量显著低于UK1;tcdB的表达以及B毒素在两种菌株的培养上清液中和声波处理的细胞内的差异都不大。结论中国艰难梭菌A-B+高分离株BJ08和欧洲流行A+B+高毒株UK1的B毒素的转录和表达没有明显差异,BJ08菌株在中国有爆发流行的可能。
Objective To study and compare the tcdA, tcdB transcription and toxin expression of the Chinese Clostridium difficile A-B + highly isolates BJ08 and the European popular A + B + highly virulent strain (BI / NAP1 / 027) UK1. Methods Clostridium difficile and culture supernatant were collected every 3 hours. The expression of tcdA and tcdB genes in BJ08 and UK1 strains were detected by real-time fluorescence quantitative PCR. The expression of tcdA and tcdB genes in BJ08 and UK1 strains were detected by ELISA. The culture supernatant A toxin, B toxin content. Results The growth rate of BJ08 was significantly higher than that of UK1; BJ08 did not express A toxin, but tcdA gene transcription was detected, tcdA gene expression was significantly lower than that of UK1; tcdB expression and B toxin were neutralized in culture supernatants of both strains Differences within the treated cells were not significant. Conclusions There was no significant difference in the transcription and expression of B toxin between the strains of B.cit diffusin A and BJ08 in China and those of the European strain A + B +. The BJ08 strain was prevalent in China.