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Related Concept Videos

Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence10:06

Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence

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We describe the optical imaging of mice infected with Mycobacterium tuberculosis (M. tuberculosis) using reporter enzyme fluorescence (REF). This protocol facilitates the sensitive and specific detection of M. tuberculosis in pre-clinical animal models for pathogenesis, therapeutics and vaccine...
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The MODS method for diagnosis of tuberculosis and multidrug resistant tuberculosis23:06

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This video demonstrates the reporter enzyme fluorescence (REF) imaging for in vivo detection of Mycobacterium tuberculosis infection. The infected mice are intraperitoneally injected with a substrate for REF imaging. Upon being hydrolyzed by bacterial β-lactamase enzyme, the substrate emits fluorescence, the detection of which aids evaluation of...
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Measuring Drug Binding to Caseum from Tuberculosis Lesions via the Rapid Equilibrium Dialysis Method03:00

Measuring Drug Binding to Caseum from Tuberculosis Lesions via the Rapid Equilibrium Dialysis Method

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Source: Sarathy, J. P. et al. An In Vitro Caseum Binding Assay that Predicts Drug Penetration in Tuberculosis Lesions. J. Vis. Exp. (2017)This video demonstrates an in vitro assay for measuring drug binding to caseum from tuberculosis lesions using rapid equilibrium dialysis. It outlines the steps for preparing caseum suspensions, performing dialysis, and calculating the unbound drug fraction to predict drug penetration into...
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Related Experiment Video

Updated: Jan 19, 2026

Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence
10:06

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Published on: February 26, 2018

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Rapid Tuberculosis Diagnosis Using Reporter Enzyme Fluorescence.

Preeti Sule1, Ronak Tilvawala1, Toriq Mustapha1

  • 1Department of Microbial Pathogenesis and Immunology, Texas A&M Health Science Center, Bryan, Texas, USA.

Journal of Clinical Microbiology
|September 13, 2019
PubMed
Summary

A new rapid diagnostic test for tuberculosis (TB) offers quick results in 10 minutes. This reporter enzyme fluorescence assay (REFtb) shows high sensitivity and specificity, potentially improving early diagnosis and reducing disease spread.

Keywords:
BlaCMycobacterium tuberculosisbiomarkerlactamasepoint of care

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Last Updated: Jan 19, 2026

Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence
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The MODS method for diagnosis of tuberculosis and multidrug resistant tuberculosis
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Area of Science:

  • Microbiology
  • Infectious Diseases
  • Biotechnology

Background:

  • Tuberculosis (TB) remains a leading cause of infectious disease mortality worldwide.
  • Current diagnostic methods often lead to delayed diagnosis due to low bacterial loads in early stages.
  • Delayed diagnosis contributes to the continued spread of TB.

Purpose of the Study:

  • To evaluate a novel, rapid diagnostic assay for tuberculosis (TB).
  • To assess the performance of the reporter enzyme fluorescence assay (REFtb) using clinical specimens.
  • To compare REFtb assay results against established TB diagnostic standards.

Main Methods:

  • Development of a rapid diagnostic assay, REFtb, utilizing the Mycobacterium tuberculosis biomarker BlaC.
  • Optimization of the assay with a specific fluorogenic substrate, CDG-3, for clinical samples.
  • Evaluation of REFtb assay performance on clinical specimens from 160 patients across 16 countries, comparing with culture and smear microscopy.

Main Results:

  • The REFtb assay demonstrated a sensitivity of 89% for smear-positive, culture-positive TB samples and 88% for smear-negative, culture-positive samples.
  • The specificity of the REFtb assay was determined to be 82%.
  • The assay achieved a negative predictive value of 93% and a positive predictive value of 79% for TB infection.

Conclusions:

  • The REFtb assay shows promise as a rapid and sensitive diagnostic tool for tuberculosis.
  • Further independent validation studies are recommended to confirm the accuracy and clinical utility of a commercial REFtb kit.
  • Successful implementation of REFtb could significantly improve early case identification and reduce TB transmission.