Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay
We used a colorimetric reverse dot blot hybridization (CRDH) assay to detect the presence of mutations in a specific region of the rpoB gene, associated with rifampin (RIF) resistance, in a panel of 156 DNAs extracted from 103 RIF-sensitive and 53 RIF-resistant cultures of Mycobacterium tuberculosis...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Fundação Oswaldo Cruz (FIOCRUZ)
2011-03-01
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| Series: | Memorias do Instituto Oswaldo Cruz |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762011000200004&lng=en&tlng=en |
| _version_ | 1797756592088678400 |
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| author | Raquel de A Maschmann Mirela Verza Marcia SN Silva Rosa Dea Sperhacke Marta O Ribeiro Philip Noel Suffys Harrison Magdinier Gomes Enrico Tortoli Fiorella Marcelli Arnaldo Zaha Maria Lucia R Rossetti |
| author_facet | Raquel de A Maschmann Mirela Verza Marcia SN Silva Rosa Dea Sperhacke Marta O Ribeiro Philip Noel Suffys Harrison Magdinier Gomes Enrico Tortoli Fiorella Marcelli Arnaldo Zaha Maria Lucia R Rossetti |
| author_sort | Raquel de A Maschmann |
| collection | DOAJ |
| description | We used a colorimetric reverse dot blot hybridization (CRDH) assay to detect the presence of mutations in a specific region of the rpoB gene, associated with rifampin (RIF) resistance, in a panel of 156 DNAs extracted from 103 RIF-sensitive and 53 RIF-resistant cultures of Mycobacterium tuberculosis. When compared with the antimicrobial susceptibility test (AST), the sensitivity and specificity of the CRDH were 92.3% and 98.1%, respectively. When compared with sequencing, the sensitivity and specificity of the CRDH were 90.6% and 100%, respectively. To evaluate the performance of the assay directly in clinical specimens, 30 samples from tuberculosis patients were used. For these samples, the results of the CRDH were 100% consistent with the results of the AST and sequencing. These results indicate that the rate of concordance of the CRDH is high when compared to conventional methods and sequencing data. The CRDH can be successfully applied when a rapid test is required for the identification of RIF resistance in M. tuberculosis. |
| first_indexed | 2024-03-12T18:03:47Z |
| format | Article |
| id | doaj.art-b8c46a3f198841f8bf86b00a660bc032 |
| institution | Directory Open Access Journal |
| issn | 1678-8060 |
| language | English |
| last_indexed | 2024-03-12T18:03:47Z |
| publishDate | 2011-03-01 |
| publisher | Fundação Oswaldo Cruz (FIOCRUZ) |
| record_format | Article |
| series | Memorias do Instituto Oswaldo Cruz |
| spelling | doaj.art-b8c46a3f198841f8bf86b00a660bc0322023-08-02T09:32:30ZengFundação Oswaldo Cruz (FIOCRUZ)Memorias do Instituto Oswaldo Cruz1678-80602011-03-01106213914510.1590/S0074-02762011000200004S0074-02762011000200004Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assayRaquel de A Maschmann0Mirela Verza1Marcia SN Silva2Rosa Dea Sperhacke3Marta O Ribeiro4Philip Noel Suffys5Harrison Magdinier Gomes6Enrico Tortoli7Fiorella Marcelli8Arnaldo Zaha9Maria Lucia R Rossetti10Fundação Estadual de Produção e Pesquisa em SaúdeFundação Estadual de Produção e Pesquisa em SaúdeFundação Estadual de Produção e Pesquisa em SaúdeUniversidade de Caxias do SulFundação Estadual de Produção e Pesquisa em SaúdeFundação Oswaldo CruzFundação Oswaldo CruzCentro Regionale di Riferimento per i MicobatteriCentro Regionale di Riferimento per i MicobatteriUniversidade Federal do Rio Grande do SulFundação Estadual de Produção e Pesquisa em SaúdeWe used a colorimetric reverse dot blot hybridization (CRDH) assay to detect the presence of mutations in a specific region of the rpoB gene, associated with rifampin (RIF) resistance, in a panel of 156 DNAs extracted from 103 RIF-sensitive and 53 RIF-resistant cultures of Mycobacterium tuberculosis. When compared with the antimicrobial susceptibility test (AST), the sensitivity and specificity of the CRDH were 92.3% and 98.1%, respectively. When compared with sequencing, the sensitivity and specificity of the CRDH were 90.6% and 100%, respectively. To evaluate the performance of the assay directly in clinical specimens, 30 samples from tuberculosis patients were used. For these samples, the results of the CRDH were 100% consistent with the results of the AST and sequencing. These results indicate that the rate of concordance of the CRDH is high when compared to conventional methods and sequencing data. The CRDH can be successfully applied when a rapid test is required for the identification of RIF resistance in M. tuberculosis.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762011000200004&lng=en&tlng=endrug resistancereverse dot blot hybridizationrifampinrpoBtuberculosis |
| spellingShingle | Raquel de A Maschmann Mirela Verza Marcia SN Silva Rosa Dea Sperhacke Marta O Ribeiro Philip Noel Suffys Harrison Magdinier Gomes Enrico Tortoli Fiorella Marcelli Arnaldo Zaha Maria Lucia R Rossetti Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay Memorias do Instituto Oswaldo Cruz drug resistance reverse dot blot hybridization rifampin rpoB tuberculosis |
| title | Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay |
| title_full | Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay |
| title_fullStr | Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay |
| title_full_unstemmed | Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay |
| title_short | Detection of rifampin-resistant genotypes in Mycobacterium tuberculosis by reverse hybridization assay |
| title_sort | detection of rifampin resistant genotypes in mycobacterium tuberculosis by reverse hybridization assay |
| topic | drug resistance reverse dot blot hybridization rifampin rpoB tuberculosis |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762011000200004&lng=en&tlng=en |
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