A cytoderm metabolic labeling TPAPy-Tre for real-time detection of vitality of Mycobacterium tuberculosis in sputum

ABSTRACT Early bactericidal activity is a vital measure in developing new tuberculosis (TB) drugs. Traditional methods, including colony-forming units (CFUs) and time to positivity (TTP), have limitations. This study aimed to evaluate the efficacy of TPAPy-Tre fluorescence microscopy in monitoring e...

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Main Authors: Mengru Yang, Guiqin Dai, Dan Li, Pengfei Zhao, Senlin Zhan, Hongjuan Qin, Hongzhou Lu, Mingbin Zheng, Peize Zhang
Format: Article
Language:English
Published: American Society for Microbiology 2025-07-01
Series:Microbiology Spectrum
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Online Access:https://journals.asm.org/doi/10.1128/spectrum.02457-24
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author Mengru Yang
Guiqin Dai
Dan Li
Pengfei Zhao
Senlin Zhan
Hongjuan Qin
Hongzhou Lu
Mingbin Zheng
Peize Zhang
author_facet Mengru Yang
Guiqin Dai
Dan Li
Pengfei Zhao
Senlin Zhan
Hongjuan Qin
Hongzhou Lu
Mingbin Zheng
Peize Zhang
author_sort Mengru Yang
collection DOAJ
description ABSTRACT Early bactericidal activity is a vital measure in developing new tuberculosis (TB) drugs. Traditional methods, including colony-forming units (CFUs) and time to positivity (TTP), have limitations. This study aimed to evaluate the efficacy of TPAPy-Tre fluorescence microscopy in monitoring early therapeutic responses compared with conventional culture methods in patients with pulmonary multidrug-resistant/rifampicin-resistant TB. In an open-label clinical trial at the Third People’s Hospital of Shenzhen, China, seven sputum smear-positive patients aged ≥18 years were enrolled. Sputum samples were consecutively analyzed using solid and liquid cultures and TPAPy-Tre microscopy. The study found that TPAPy-Tre fluorescence intensity significantly decreased from 271.5 (95% confidence interval [CI], 177.4–365.7) before treatment to 142.8 (95% CI, 104.7–180.9) after treatment (P < 0.05). TPAPy-Tre results strongly correlated with CFU (Spearman ρ = 0.60; 95% CI, 0.35–0.77; P < 0.001) and TTP (Spearman ρ = −0.33; 95% CI, −0.56 to −0.04; P < 0.05). Among selected participants, the median fluorescence intensity decreased from 51.5 (interquartile range [IQR], 39.0–63.6) to 13.2 (IQR, 7.8–20.0) after treatment (P < 0.001). TPAPy-Tre shows potential as a rapid, visual method for tracking bacterial vitality during TB treatment, offering immediate feedback on treatment response. These results support its use alongside conventional methods in clinical settings, though larger studies are needed for further validation.IMPORTANCEEarly bactericidal activity (EBA) is an important tool in clinical studies in the development of new tuberculosis drugs. Current traditional methods of efficacy monitoring present significant limitations. There is a need for novel and efficient tools to monitor treatment response in real-time when EBA is performing.
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issn 2165-0497
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publishDate 2025-07-01
publisher American Society for Microbiology
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series Microbiology Spectrum
spelling doaj-art-df4adbbb3b894ab8be4464946d408f162025-08-20T03:28:22ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-07-0113710.1128/spectrum.02457-24A cytoderm metabolic labeling TPAPy-Tre for real-time detection of vitality of Mycobacterium tuberculosis in sputumMengru Yang0Guiqin Dai1Dan Li2Pengfei Zhao3Senlin Zhan4Hongjuan Qin5Hongzhou Lu6Mingbin Zheng7Peize Zhang8School of Public Health, Shenzhen University Medical School, Shenzhen University, Shenzhen, Guangdong, ChinaInstitute of Hepatology, National Clinical Research Center for Infectious Diseases, Third People’s Hospital of Shenzhen, Shenzhen, Guangdong, ChinaNational Clinical Research Center for Infectious Diseases, Shenzhen Clinical Research Center for Tuberculosis, Shenzhen Third People’s Hospital, Shenzhen, Guangdong, ChinaInstitute of Hepatology, National Clinical Research Center for Infectious Diseases, Third People’s Hospital of Shenzhen, Shenzhen, Guangdong, ChinaDepartment of Pulmonary Medicine and Tuberculosis, Third People’s Hospital of Shenzhen, Shenzhen, Guangdong, ChinaDepartment of Pulmonary Medicine and Tuberculosis, Third People’s Hospital of Shenzhen, Shenzhen, Guangdong, ChinaDepartment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Third People’s Hospital of Shenzhen, Shenzhen, Guangdong, ChinaShenzhen Clinical Research Center for Tuberculosis, Shenzhen, ChinaSchool of Public Health, Shenzhen University Medical School, Shenzhen University, Shenzhen, Guangdong, ChinaABSTRACT Early bactericidal activity is a vital measure in developing new tuberculosis (TB) drugs. Traditional methods, including colony-forming units (CFUs) and time to positivity (TTP), have limitations. This study aimed to evaluate the efficacy of TPAPy-Tre fluorescence microscopy in monitoring early therapeutic responses compared with conventional culture methods in patients with pulmonary multidrug-resistant/rifampicin-resistant TB. In an open-label clinical trial at the Third People’s Hospital of Shenzhen, China, seven sputum smear-positive patients aged ≥18 years were enrolled. Sputum samples were consecutively analyzed using solid and liquid cultures and TPAPy-Tre microscopy. The study found that TPAPy-Tre fluorescence intensity significantly decreased from 271.5 (95% confidence interval [CI], 177.4–365.7) before treatment to 142.8 (95% CI, 104.7–180.9) after treatment (P < 0.05). TPAPy-Tre results strongly correlated with CFU (Spearman ρ = 0.60; 95% CI, 0.35–0.77; P < 0.001) and TTP (Spearman ρ = −0.33; 95% CI, −0.56 to −0.04; P < 0.05). Among selected participants, the median fluorescence intensity decreased from 51.5 (interquartile range [IQR], 39.0–63.6) to 13.2 (IQR, 7.8–20.0) after treatment (P < 0.001). TPAPy-Tre shows potential as a rapid, visual method for tracking bacterial vitality during TB treatment, offering immediate feedback on treatment response. These results support its use alongside conventional methods in clinical settings, though larger studies are needed for further validation.IMPORTANCEEarly bactericidal activity (EBA) is an important tool in clinical studies in the development of new tuberculosis drugs. Current traditional methods of efficacy monitoring present significant limitations. There is a need for novel and efficient tools to monitor treatment response in real-time when EBA is performing.https://journals.asm.org/doi/10.1128/spectrum.02457-24early bactericidal activity (EBA)tuberculosis (TB)TPAPy-Tre microscopyfluorescence intensitytreatment response monitoring
spellingShingle Mengru Yang
Guiqin Dai
Dan Li
Pengfei Zhao
Senlin Zhan
Hongjuan Qin
Hongzhou Lu
Mingbin Zheng
Peize Zhang
A cytoderm metabolic labeling TPAPy-Tre for real-time detection of vitality of Mycobacterium tuberculosis in sputum
Microbiology Spectrum
early bactericidal activity (EBA)
tuberculosis (TB)
TPAPy-Tre microscopy
fluorescence intensity
treatment response monitoring
title A cytoderm metabolic labeling TPAPy-Tre for real-time detection of vitality of Mycobacterium tuberculosis in sputum
title_full A cytoderm metabolic labeling TPAPy-Tre for real-time detection of vitality of Mycobacterium tuberculosis in sputum
title_fullStr A cytoderm metabolic labeling TPAPy-Tre for real-time detection of vitality of Mycobacterium tuberculosis in sputum
title_full_unstemmed A cytoderm metabolic labeling TPAPy-Tre for real-time detection of vitality of Mycobacterium tuberculosis in sputum
title_short A cytoderm metabolic labeling TPAPy-Tre for real-time detection of vitality of Mycobacterium tuberculosis in sputum
title_sort cytoderm metabolic labeling tpapy tre for real time detection of vitality of mycobacterium tuberculosis in sputum
topic early bactericidal activity (EBA)
tuberculosis (TB)
TPAPy-Tre microscopy
fluorescence intensity
treatment response monitoring
url https://journals.asm.org/doi/10.1128/spectrum.02457-24
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