Development and evaluation of a single-tube nested PCR with colorimetric assay for Mycobacterium tuberculosis detection.
Marcela Pereira SalazarJuliana Figueiredo da Costa Lima Suassuna MonteiroWlisses Henrique Veloso Carvalho-SilvaGeorge Tadeu Nunes DinizRoberto Pereira WerkhauserLílian Maria Lapa MontenegroHaiana Charifker SchindlerPublished in: BioTechniques (2024)
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.