Development and evaluation of a single-tube nested PCR with colorimetric assay for Mycobacterium tuberculosis detection

Development and evaluation of a single-tube nested PCR with colorimetric assay for Mycobacterium tuberculosis detection

Molecular techniques have revolutionized tuberculosis (TB) diagnosis by offering a faster and more sensitive approach, detecting Mycobacterium tuberculosis (Mtb) DNA directly from samples. Single-tube nested PCR (STNPCR) combines two PCR reactions with separate oligonucleotide sets in a single tube....

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Main Authors: Marcela Pereira Salazar, Juliana Figueiredo da Costa Lima Suassuna Monteiro, Wlisses Henrique Veloso Carvalho-Silva, George Tadeu Nunes Diniz, Roberto Pereira Werkhauser, Lílian Maria Lapa Montenegro, Haiana Charifker Schindler
Format: Article
Language:English
Published: Taylor & Francis Group 2024-06-01
Series:BioTechniques
Subjects:
IS6110
Mtb complex
rapid molecular test
STNPCR
tuberculosis diagnosis
Online Access:https://www.tandfonline.com/doi/10.2144/btn-2023-0080
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Summary:Molecular techniques have revolutionized tuberculosis (TB) diagnosis by offering a faster and more sensitive approach, detecting Mycobacterium tuberculosis (Mtb) DNA directly from samples. Single-tube nested PCR (STNPCR) combines two PCR reactions with separate oligonucleotide sets in a single tube. Moreover, colorimetric methods in PCR products have been studied for pathogen detection. Thus, this study aimed to establish a novel system based on colorimetric STNPCR for Mtb detection using microtiter plates with IS6110-amplified fragments. The results showed a general colorimetric STNPCR detection limit of 1 pg/μl. Its general sensitivity and specificity were 76.62 and 60.53%, respectively, with kappa index agreement of 0.166.
ISSN:0736-6205
1940-9818

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