2. Academic Validation
  3. Real-time bioluminescence imaging of mycobacteria with Akaluc: a novel method for monitoring drug efficacy

Real-time bioluminescence imaging of mycobacteria with Akaluc: a novel method for monitoring drug efficacy

  • Sci Rep. 2026 May 15;16(1):15193. doi: 10.1038/s41598-026-44744-6.
Md Shafiul Islam 1,  Atsuki Takeishi 2,  Yoshitaka Tateishi 3,  Akihito Nishiyama 2,  Yuriko Ozeki 2,  Yutaka Yoshida 2,  Amina Kaboso Shaban 2,  Takeshi Annoura 4 5,  Satoshi Iwano 6,  Takasuke Fukuhara 7,  Sohkichi Matsumoto 8 9 10 11
Affiliations

Affiliations

  • 1 Department of Bacteriology, Graduate School of Medical and Dental Sciences, Niigata University, 757 Ichibancho, Asahimachi-dori, Chuo-Ku, Niigata, 951-8510, Japan. [email protected].
  • 2 Department of Bacteriology, Graduate School of Medical and Dental Sciences, Niigata University, 757 Ichibancho, Asahimachi-dori, Chuo-Ku, Niigata, 951-8510, Japan.
  • 3 Department of Microbiology, Fukushima Medical University, 1Hikariga-oka, , Fukushima City, 960-1295, Japan.
  • 4 Department of Parasitology, National Institute of Infectious Diseases (NIID), Japan Institute for Health Security (JIHS), Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan.
  • 5 Cooperative Division of Veterinary Sciences, Graduate School of Agriculture, Tokyo University of Agriculture and Technology (TUAT), Tokyo, 183-8538, Japan.
  • 6 Institute for Tenure Track Promotion, University of Miyazaki, Miyazaki, Miyazaki, 889-2192, Japan.
  • 7 Department of Virology, Faculty of Medical Sciences, Kyushu University, Fukuoka, Japan.
  • 8 Department of Bacteriology, Graduate School of Medical and Dental Sciences, Niigata University, 757 Ichibancho, Asahimachi-dori, Chuo-Ku, Niigata, 951-8510, Japan. [email protected].
  • 9 Department of Bacteriology, Osaka Metropolitan University Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-ku, Osaka, 545-8586, Japan. [email protected].
  • 10 Division of Research Aids, Hokkaido University Institute for Vaccine Research and Development, Kita 20, Nishi 10, Kita-Ku, Sapporo, Hokkaido, 001-0020, Japan. [email protected].
  • 11 Laboratory of Tuberculosis, Institute of Tropical Disease, Universitas Airlangga, Kampus C JI. Mulyorejo, Surabaya, 60113, Indonesia. [email protected].
PMID: 42151204 DOI: 10.1038/s41598-026-44744-6
Abstract

Tuberculosis (TB) still threatens human life despite the availability of childhood vaccination and modern treatment regimens due to the emergence of tuberculosis with extended resistance profiles. The mouse active TB model remains the gold standard for evaluating anti-TB drugs and vaccines. However, in vivo experiments raise ethical concerns and are time-consuming. We established an Akaluc-based bioluminescence platform to enable rapid drug screening in culture media and THP‑1 cells. Codon-optimized Akaluc expression in mycobacteria was accomplished by assortment of vectors and promoters. Bioluminescence kinetics were evaluated in culture media and THP‑1 cells with or without drug treatment, and optimized by adjusting time points and substrate concentrations. TokeOni at a concentration of 10 nM/100 µL produced the highest bioluminescence compared to Other tested concentrations and substrates. Among the tested promoter-plasmid constructs, the Ag85B promoter in pMV261 generated the strongest bioluminescence in Mycobacterium smegmatis and Mycobacterium bovis. Bioluminescence fluctuated with Bacterial growth, peaking during the log phase and gradually declining during the stationary phase. A positive correlation was observed between bioluminescence and CFU reduction in vitro upon treatment with sensitive drugs.

Keywords

Akaluc; Antigen 85B (Ag85B); Bioluminescence imaging; Drug efficacy; TokeOni; Tuberculosis.

Figures
Products