Rapid detection of Mycobacterium tuberculosis DNA and genetic markers for Isoniazid resistance in Ziehl-Neelsen stained slides
| Autor: | Mirela Gehlen, Maria Lucia Rosa Rossetti, Isabela Neves de Almeida, Elis Regina Dalla Costa, Sheile Pinheiro de Jesus, Franciele Costa Leite Morais, Jonas Michel Wolf, Lida Jouca de Assis Figueiredo, Silvana Spíndola de Miranda, Graziele Lima Bello, Tainá dos Santos Soares, Regina Bones Barcellos |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Microbiology (medical)
DNA Bacterial Genetic Markers Tuberculosis Genotype 030231 tropical medicine RC955-962 Antitubercular Agents Drug resistance Microbial Sensitivity Tests Real-Time Polymerase Chain Reaction Sensitivity and Specificity Microbiology law.invention Mycobacterium tuberculosis 03 medical and health sciences 0302 clinical medicine law Arctic medicine. Tropical medicine Drug Resistance Bacterial medicine Isoniazid Humans Genotyping Polymerase chain reaction biology business.industry INHA medicine.disease biology.organism_classification Molecular biology Isoniazid resistance QR1-502 genotyping Mutation Ziehl–Neelsen stain Original Article business real-time PCR medicine.drug |
| Zdroj: | Memórias do Instituto Oswaldo Cruz, Volume: 115, Article number: e190407, Published: 17 APR 2020 Memórias do Instituto Oswaldo Cruz., Vol 115 (2020) Memórias do Instituto Oswaldo Cruz |
| Popis: | BACKGROUND Early diagnosis of tuberculosis (TB) and identification of strains of Mycobacterium tuberculosis resistant to anti-TB drugs are considered the main factors for disease control. OBJECTIVES To standardise a real-time polymerase chain reaction (qPCR) assay technique and apply it to identify mutations involved in M. tuberculosis resistance to Isoniazid (INH) directly in Ziehl-Neelsen (ZN) stained slides. METHODS Were analysed 55 independent DNA samples extracted from clinical isolates of M. tuberculosis by sequencing. For application in TB diagnosis resistance, 59 ZN-stained slides were used. The sensitivity, specificity and Kappa index, with a 95% confidence interval (CI95%), were determined. FINDINGS The agreement between the tests was, for the katG target, the Kappa index of 0.89 (CI95%: 0.7-1.0). The sensitivity and specificity were 97.6% (CI95%: 87.7-99.9) and 91.7% (CI95%: 61.5-99.5), respectively. For inhA, the Kappa index was 0.92 (CI95%: 0.8-1.0), the sensitivity and specificity were 94.4% (CI95%: 72.7-99.8) and 97.3% (CI95%: 85.8-99.9), respectively. The use of ZN-stained slides for drug-resistant TB detection showed significant results when compared to other standard tests for drug resistance. MAIN CONCLUSIONS qPCR genotyping proved to be an efficient method to detect genes that confer M. tuberculosis resistance to INH. Thus, qPCR genotyping may be an alternative instead of sequencing. |
| Databáze: | OpenAIRE |
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