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Published on: September 14, 2011
Application of BactTiter-Glo ATP bioluminescence assay for Mycobacterium tuberculosis detection
Mariana Abou Mourad Ferreira1, Laura Candeias Dos Santos1, Luiz Guilherme Schmidt Castellani1
1Núcleo de Doenças Infecciosas, Centro de Ciências da Saúde (Health Sciences Center), Universidade Federal do Espírito Santo, Av. Marechal Campos, 1468, 29040-090 Vitória, Espírito Santo, Brazil.
Background:
Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), remains a global health threat, necessitating faster and more accessible diagnostic methods. This study investigates critical parameters in the application of a commercial ATP bioluminescence assay for the detection of MTB.
Method:
Our objective was to optimize the ATP bioluminescence protocol using BacTiter-Glo™ for MTB, investigating the impact of varying volumes of MTB suspension and reagent on assay sensitivity, evaluating ATP extraction methods, establishing calibration curves, and elucidating strain-specific responses to antimicrobial agents.
Results:
ATP extraction methods showed no significant improvement over controls. Calibration curves revealed a linear correlation between relative light units (RLU) and colony-forming units (CFU/mL), establishing low detection limits. Antimicrobial testing demonstrated strain-specific responses aligning with susceptibility and resistance patterns.
Conclusion:
Our findings contribute to refining ATP bioluminescence protocols for enhanced MTB detection and susceptibility testing. Further refinements and validation efforts are warranted, holding promise for more efficient diagnostic platforms in the future.
