生孢梭菌CFU计数培养基的研制  

Development of CFU Counting Culture Medium for Clostridium sporogenes

作　　者：高尚先[1] 孙彬裕[1] 康国华[1] 曲守方[1] 

机构地区：[1]中国药品生物制品检定所,北京100050

出　　处：《药物分析杂志》2005年第1期11-13,共3页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:研制一种适宜生孢子梭状芽孢杆菌(简称生孢梭菌)菌落形成单位(Colony Forming Units,CFU)计数的微生物培养基(简称培养基),使其CFU计数能够接近最大活菌数。方法:将生孢梭菌接种于硫乙醇酸盐培养基(不含琼脂),置35℃培养24-48h;将生孢梭菌的新鲜培养物置无菌离心管中,500r·min^(-1)离心5min。取上层菌液,用0.9%无菌氯化钠溶液稀释至与标准比浊管相同浓度,并作10倍系列稀释至1×10^(-6)备用。将上述菌液分别浇注混匀于琼脂浓度为0.5%,0.6%和0.7%的3个生孢梭菌CFU计数研究培养基(简称3个生孢梭菌CFU计数研究培养基),置厌氧条件下35℃培养。优选18h进行CFU/mL计数并观察菌落形态,优选琼脂浓度为0.5%的上述培养基作为生孢梭菌CFU计数培养基并与其他5个培养基进行生孢梭菌CFU计数的比较试验。将上述菌液采用浇注法和L棒涂抹法接种于上述6个不同的培养基,置不同培养条件及培养时间进行生孢梭菌的CFU计数。结果:生孢梭菌CFU计数培养基,浇注法接种,在厌氧条件下35℃培养18h,CFU计数最多,且菌落形态良好,可能最接近最大活菌数。结论:新研制的生孢梭菌CFU计数培养基是目前最适于生孢梭菌CFU计数的培养基。Objective:To develop a kind of appropriate CFU (Colony Forming Units) counting culture medium for Clostridium sporogenes, make CFU Count be approximate to the most amount of practical alive bacteria. Methods:Inoculated Clostridium sporogenes on fluid thioglycollate culture medium(without agar). Incubated the medium inoculated with Clostridium sporogenes for 24-48 h at 35℃;put the fresh medium with Clostridium sporogenes into centrifugal tube,at 500 r·min^(-1) for 5 min. Took the upper bacterial suspension and diluted it with 0.9% sterilized NaCl solution to a concentration against the standard opacity, and then carried out 10-fold serial dilutions to the dilution of 1×10^(-6). Put the bacterial suspension into 3 CFU counting researching culture media (with 0.5% ,0.6% and 0.7% agar respectively)for Clostridium sporogenes with pouling method under anaerobic condition at 35℃. Preferred to choose 18 h for CFU Count and colony observation, and to choose the researching medium with 0.5% agar as the CFU counting culture medium for Clostridium sporogenes compared with other 5 media. Inoculated the above bacterial suspension into the 6 different media with pouling method and L stick daubing method for CFU Count under different culture conditions and culture temperature. Results:When inoculated Clostridium sporogenes with pouring method into the CFU counting culture medium for Clostridium sporogene for 18 h at 35℃ under anaerobic condition, CFU Count was the most and colony configuration was good. We thought it may be the most approximate to the most amount of practical alive bacteria. Conclusions:In the present, the new developing medium may be the most appropriate medium for Clostridium sporogenes to count CFU.

关 键 词：梭菌 培养基 CFU 琼脂浓度 菌液 接种 稀释 计数 无菌 离心管 

分 类 号：R978.1[医药卫生—药品] S858[医药卫生—药学]