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

Biotechniques. 2024 Apr 11. doi: 10.2144/btn-2023-0080. Online ahead of print.ABSTRACTMolecular 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.PMID:38602382 | DOI:10.2144/btn-2023-0080

Source: BioTechniques - April 11, 2024 Category: Biotechnology Authors: Marcela Pereira Salazar Juliana Figueiredo da Costa Lima Suassuna Monteiro Wlisses Henrique Veloso Carvalho-Silva George Tadeu Nunes Diniz Roberto Pereira Werkhauser L ílian Maria Lapa Montenegro Haiana Charifker Schindler Source Type: research