Optimizing a high-sensitivity NanoLuc-based bioluminescence system for in vivo evaluation of antimicrobial treatment  

作　　者：Weilong Shang Zhen Hu Mengyang Li Yuting Wang Yifan Rao Li Tan Juan Chen Xiaonan Huang Lu Liu He Liu Zuwen Guo Huagang Peng Yi Yang Qiwen Hu Shu Li Xiaomei Hu Jiao Zou Xiancai Rao 

机构地区：[1]Department of Microbiology,College of Basic Medical Sciences,Key Laboratory of Microbial Engineering Under the Educational Committee in Chongqing,Army Medical University(Third Military Medical University),Chongqing,China [2]Department of Microbiology,School of Medicine,Chongqing University,Chongqing,China [3]Department of Emergency Medicine,Xinqiao Hospital,Army Medical University(Third Military Medical University),Chongqing,China [4]Department of Pharmacy,Xinqiao Hospital,Army Medical University(Third Military Medical University),Chongqing,China [5]Department of Military Cognitive Psychology,School of Psychology,Army Medical University(Third Military Medical University),Chongqing,China.

出　　处：《mLife》2023年第4期462-478,共17页微生物（英文）

基　　金：supported by the National Natural Science Foundation of China (No.82072238 to W.S.and 82272341 to X.R.).

摘　　要：Focal and systemic infections are serious threats to human health.Preclinical models enable the development of new drugs and therapeutic regimens.In vivo,animal bioluminescence(BL)imaging has been used with bacterial reporter strains to evaluate antimicrobial treatment effects.However,high-sensitivity bioluminescent systems are required because of the limited tissue penetration and low brightness of the BL signals of existing approaches.Here,we report that NanoLuc(Nluc)showed better performance than LuxCDABE in bacteria.However,the retention rate of plasmid constructs in bacteria was low.To construct stable Staphylococcus aureus reporter strains,a partner protein enolase(Eno)was identified by screening of S.aureus strain USA300 for fusion expression of Nluc-based luciferases,including Nluc,Teluc,and Antares2.Different substrates,such as hydrofurimazine(HFZ),furimazine(FUR),and diphenylterazine(DTZ),were used to optimize a stable reporter strain/substrate pair for BL imaging.S.aureus USA300/Eno-Antares2/HFZ produced the highest number of photons of orange-red light in vitro and enabled sensitive BL tracking of S.aureus in vivo,with sensitivities of approximately 10 CFU from mouse skin and 750 CFU from mouse kidneys.USA300/Eno-Antares2/HFZ was a powerful combination based on the longitudinal evaluation of the therapeutic efficacy of antibiotics.The optimized S.aureus Eno-Antares2/HFZ pair provides a technological advancement for the in vivo evaluation of antimicrobial treatment.

关 键 词：antimicrobial evaluation bioluminescence imaging Nluc-based luciferases Staphylococcus aureus therapeutic efficacy 

分 类 号：R37[医药卫生—病原生物学]