Uemura T, Sakaguchi G, Riemann H P
Appl Microbiol. 1973 Sep;26(3):381-5. doi: 10.1128/am.26.3.381-385.1973.
The reversed passive hemagglutination (RPHA) test yielded a positive reaction in 2 h with as little as 0.5 ng of purified Clostridium perfringens enterotoxin (CPE) per ml as well as with cultures of some C. perfringens grown in Duncan-Strong (DS) medium. This method is the most sensitive, the simplest, and the fastest among all reported. The time course of CPE production of Clostridium perfringens NCTC 8798 in DS was investigated by RPHA. CPE in culture was detectable at 4 h, increased gradually, reached a maximum at 12 to 14 h, and remained at a high level of 20 mug/ml through 48 h of incubation. CPE synthesized within cells is released easily by sonic disruption of young cultures and by aging the cultures 20 h or more. Heat shock of the cell inoculum was essential for CPE production by C. perfringens in DS.
反向被动血凝试验(RPHA)在2小时内对每毫升低至0.5纳克的纯化产气荚膜梭菌肠毒素(CPE)以及在邓肯-斯特朗(DS)培养基中生长的一些产气荚膜梭菌培养物产生阳性反应。该方法是所有报道中最灵敏、最简单且最快的。通过RPHA研究了产气荚膜梭菌NCTC 8798在DS中产生CPE的时间进程。培养物中的CPE在4小时时可检测到,逐渐增加,在12至14小时达到最大值,并在48小时的孵育过程中保持在20微克/毫升的高水平。细胞内合成的CPE通过对年轻培养物进行超声破碎以及使培养物老化20小时或更长时间可轻易释放。细胞接种物的热休克对于产气荚膜梭菌在DS中产生CPE至关重要。