香港海鸥菌不同保存方法效果比较  

Comparison of methods for Laribacter hongkongensis strain storage

作　　者：刘优招 冯梅[1] 王丽[1] 陈炜聪 韩宇宇 王祉蕴 陈清[1] 刘志华[4] 胡静[1] LIU You-zhao FENG Mei WANG Li et al(Department of Epidemiology, School of Public Health, Southern Medical University, Guangzhou , Guangdong Province 510515, China)

机构地区：[1]南方医科大学流行病学系,广东广州510515 [2]南方医科大学中医药学院 [3]深圳罗湖区疾病预防与控制中心 [4]南方医院感染内科

出　　处：《中国公共卫生》2017年第3期471-474,共4页Chinese Journal of Public Health

基　　金：国家自然科学基金(81373052)

摘　　要：目的比较不同方法对香港海鸥菌的短期和长期保存效果,以期建立该菌的简单、实用和经济的短期和长期保存方法。方法采用半固体石蜡封存法,磁珠菌种保存管、10%甘油、15%甘油肉汤和30%甘油肉汤等5种方法保存香港海鸥菌,其中,半固体石蜡封存法的保存温度为室温和4℃,其他方法均在-20℃和-80℃进行菌种保存。定期进行菌种复苏,采用活菌菌落计数比较不同方法的保存效果,观察期为12个月。结果半固体石蜡封存法在室温环境下保存的活菌数在6个月内呈缓慢下降趋势(活菌数的对数值下降幅度<1),但6个月以后活菌数急速下降且菌落变小,至第9个月时为0 CFU/mL;而4℃保存的第1个月活菌数下降幅度小,但到第3个月时活菌数为0 CFU/mL;在12个月内,磁株及3种不同甘油浓度保存的活菌数在-80℃下保持相对稳定,但-20℃下保存时间短、效果差。结论室温条件下的半固体石蜡封存法适合短期保存香港海鸥菌,而-80℃低温条件下的磁株或甘油适合长期保存。Objective To compare different methods for preserving Laribacter hongkongensis （LH） strains and to develop a simple and practicable preservation protocol for short- and long-term storage and reactivation LH strain.Methods The five methods for LH strain storage included：（i） semisolid with paraffin storage,（ii） Microbank vial,（iii） 10% glycerol,（iv） 15% glycerol/85% Luria-Bertani （LB） broth culture,and （v） 30% glycerol/70% LB broth culture.The LH strains were put at 4 ℃ and room temperature when using methods i,while the stains were disposed at conventional freezing temperatures of -20 ℃ and -80 ℃ when using other methods,respectively.All storage techniques were evaluated periodically by counting with colony-forming unit over 12 months.Results For semisolid with paraffin storage,the viable strains were reduced gently up to 6 months at room temperature,then reduced rapidly and LH colonies became pinpoint size,contrasting with no colony growth on MacConkey agar at 3-month at 4 ℃.Preservation of LH strains in Microbank vial,10% glycerol,15% glycerol broth,and 30% glycerol broth stored at -80 ℃ were proved to be stable for up to 12 months.However,recovery of the strains from these methods stored at -20 ℃ showed poor growth after one month.Conclusion Semisolid with paraffin storage at room temperature is an effective way for short-term preservation of LH strain.Preservation in glycerol or Microbank vial at -80 ℃ is recommended for prolonged periods of LH strain storage.

关 键 词：香港海鸥菌 保存 活菌菌落计数 甘油 

分 类 号：R181.24[医药卫生—流行病学]