Jiang Zhi-Dong, Alexander Ashley, Ke Shi, Valilis Evangelia M, Hu Shaofan, Li Bingjie, DuPont Herbert L
Center for Infectious Diseases, Department of Epidemiology, Human Genetics & Environmental Sciences, School of Public Health, UT Health Houston, TX, USA.
Kelsey Research Foundation, Houston, TX, USA.
Anaerobe. 2017 Dec;48:110-114. doi: 10.1016/j.anaerobe.2017.08.003. Epub 2017 Aug 9.
Freezing donor fecal microbiota has simplified fecal microbiota transplantation (FMT) in the treatment of recurrent C. difficile infection (CDI). However, the optimal storage time for the frozen FMT products remains unknown. Using an established murine model of CDI, stability and efficacy of frozen and lyophilized FMT product was studied at time points from 2 months to 15 months. DNA was extracted from fecal samples from the mice with identification of specific bacterial species by real-time quantitative PCR (qPCR). FMT product stability and efficacy were measured by occurrence of diarrhea in the challenged mice together with stability of the microbiota composition. The results were analyzed and compared by SAS statistical software. All mice treated with only C. difficile developed diarrhea within 72 h. Mice treated with frozen (n = 5/group), lyophilized (n = 5/group) products stored for ≤ 7-month or fresh FMT product (n = 22) were protected from post C. difficile challenge diarrhea. There was no difference between frozen and lyophilized products (n = 5/group) stored for ≤ 7 months 95% CI 1.00 (0.38-2.64) and 1.00 (0.38-2.64), respectively. Prevention if CDI by frozen and lyophilized product was not different for storage of 9-, 11- and 15-months. qPCR results demonstrated there were no significant quantitative change in Bacteroides and Clostridium species during any of the storage times (P > 0.05). In the present study, frozen and lyophilized FMT products were stored up to 7 months without losing microbiota composition and therapeutic efficacy. The animal model described may be useful to study stability of human microbiota designed for FMT.
冷冻供体粪便微生物群简化了粪便微生物群移植（FMT）在复发性艰难梭菌感染（CDI）治疗中的应用。然而，冷冻FMT产品的最佳储存时间仍不清楚。利用已建立的CDI小鼠模型，研究了冷冻和冻干FMT产品在2个月至15个月时间点的稳定性和疗效。从小鼠粪便样本中提取DNA，通过实时定量PCR（qPCR）鉴定特定细菌种类。通过受攻击小鼠腹泻的发生情况以及微生物群组成的稳定性来衡量FMT产品的稳定性和疗效。结果采用SAS统计软件进行分析和比较。所有仅用艰难梭菌治疗的小鼠在72小时内出现腹泻。用储存≤7个月的冷冻（每组n = 5）、冻干（每组n = 5）产品或新鲜FMT产品（n = 22）治疗的小鼠在艰难梭菌攻击后可预防腹泻。储存≤7个月的冷冻和冻干产品（每组n = 5）之间没有差异，95%置信区间分别为1.00（0.38 - 2.64）和1.00（0.38 - 2.64）。冷冻和冻干产品在储存9个月、11个月和15个月时预防CDI的效果没有差异。qPCR结果表明，在任何储存时间内，拟杆菌属和梭菌属的数量均无显著变化（P > 0.05）。在本研究中，冷冻和冻干FMT产品储存长达7个月，微生物群组成和治疗效果均未丧失。所描述的动物模型可能有助于研究用于FMT的人类微生物群的稳定性。
