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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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2024-06-01
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| Series: | BioTechniques |
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| Online Access: | https://www.tandfonline.com/doi/10.2144/btn-2023-0080 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Marcela Pereira Salazar |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-4f51a26d6aa04b2f84214f5e68533f96 |
| institution | Kabale University |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2024-06-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-4f51a26d6aa04b2f84214f5e68533f962025-01-16T15:38:47ZengTaylor & Francis GroupBioTechniques0736-62051940-98182024-06-0176623524410.2144/btn-2023-0080Development and evaluation of a single-tube nested PCR with colorimetric assay for Mycobacterium tuberculosis detectionMarcela Pereira Salazar0Juliana Figueiredo da Costa Lima Suassuna Monteiro1Wlisses Henrique Veloso Carvalho-Silva2George Tadeu Nunes Diniz3Roberto Pereira Werkhauser4Lílian Maria Lapa Montenegro5Haiana Charifker Schindler6Laboratory of Immunoepidemiology, Department of Immunology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Pernambuco, 50740-465, BrazilLaboratory of Immunoepidemiology, Department of Immunology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Pernambuco, 50740-465, BrazilLaboratory of Immunoepidemiology, Department of Immunology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Pernambuco, 50740-465, BrazilLaboratory of Quantitative Methods, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Pernambuco, 50740-465, BrazilLaboratory of Immunoepidemiology, Department of Immunology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Pernambuco, 50740-465, BrazilLaboratory of Immunoepidemiology, Department of Immunology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Pernambuco, 50740-465, BrazilLaboratory of Immunoepidemiology, Department of Immunology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Pernambuco, 50740-465, BrazilMolecular 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.https://www.tandfonline.com/doi/10.2144/btn-2023-0080IS6110Mtb complexrapid molecular testSTNPCRtuberculosis diagnosis |
| spellingShingle | 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 Development and evaluation of a single-tube nested PCR with colorimetric assay for Mycobacterium tuberculosis detection BioTechniques IS6110 Mtb complex rapid molecular test STNPCR tuberculosis diagnosis |
| title | Development and evaluation of a single-tube nested PCR with colorimetric assay for Mycobacterium tuberculosis detection |
| title_full | Development and evaluation of a single-tube nested PCR with colorimetric assay for Mycobacterium tuberculosis detection |
| title_fullStr | Development and evaluation of a single-tube nested PCR with colorimetric assay for Mycobacterium tuberculosis detection |
| title_full_unstemmed | Development and evaluation of a single-tube nested PCR with colorimetric assay for Mycobacterium tuberculosis detection |
| title_short | Development and evaluation of a single-tube nested PCR with colorimetric assay for Mycobacterium tuberculosis detection |
| title_sort | development and evaluation of a single tube nested pcr with colorimetric assay for mycobacterium tuberculosis detection |
| topic | IS6110 Mtb complex rapid molecular test STNPCR tuberculosis diagnosis |
| url | https://www.tandfonline.com/doi/10.2144/btn-2023-0080 |
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