Mutations in katG and inhA genes of isoniazid-resistant and -sensitive clinical isolates of Mycobacterium tuberculosis from cases of pulmonary tuberculosis and their association with minimum inhibitory concentration of isoniazid

Jaiswal, Indu; Jain, Amita; Singh, Pooja; Verma, Sanjeev; Prakash, Shantanu; Dixit, Pratibha; Suryakant,; Singh, Mastan

doi:10.1016/j.cegh.2016.08.008

Cited by 7 publications

(6 citation statements)

References 22 publications

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“…Las mutaciones en los genes de Mycobacterium tuberculosis katG e inhA, han sido reportados como los responsables del 60 -80% de los aislados resistentes a isoniacida [27][28][29][30] .…”

Section: Discussionunclassified

Determinación de la mutación S315T del gen katG en aislados resistentes a Isoniacida de Mycobacterium tuberculosis mediante PCR-RFLP

Sánchez-Domínguez

Nicola-Salas

Morey-León

2018

Infect

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Cómo citar este artículo: J. Sánchez-Domínguez, et al. Determinación de la mutación S315T del gen katG en aislados resistentes a Isoniacida de Mycobacterium tuberculosis mediante PCR-RFLP. Infectio 2018; 22(4): 178-184 ResumenObjetivo: determinar la mutación S315T del gen katG en aislados de Mycobacterium tuberculosis resistentes a isoniacida mediante la técnica PCR-RFLP. Materiales y métodos: A partir de 68 aislados de Mycobacterium tuberculosis se realizó el análisis de polimorfismo en productos de amplificación de 1054 y 630 pb que contenían la mutación S315T del gen katG mediante PCR-RFLP empleando las enzimas de restricción MspI y SatI. Mediante SPSS se determinó sensibilidad, especificidad, valores predictivo positivo y negativo, coeficientes de probabilidad positivo y negativo. Resultados: El 74,46% de aislados fenotípicamente resistentes y 4,76% fenotípicamente sensibles presentaron la mutación del gen katG S315T. La PCR-RFLP para S315T del gen katG presentó 85,4% de sensibilidad y 95,2% de especificidad con MspI y 85,4% de sensibilidad y 94,4% de especificidad con SatI. Discusión: La PCR-RFLP tiene una alta capacidad resolutiva que depende de la enzima que se emplee como se observó en estudios previos. La presencia de la mutación S315T en pacientes vírgenes al tratamiento sugiere la circulación de aislados resistentes a Isoniacida. Conclusión: La PCR-RFLP resultó una alternativa válida y rápida para el diagnóstico de la resistencia a isoniacida, mediante la detección de la mutación S315T del gen katG en comparación con el método convencional de las proporciones.Palabras Clave: Mutaciones, Polimorfismo, Isoniacida, PCR-RFLP, Mycobacterium tuberculosis, katG. Determination of S315T mutation within the katG gene in isoniazid-resistant Mycobacterium tuberculosis isolate by PCR-RFLP AbstractObjetive: To determine the S315T mutation of the katG gene in Mycobacterium tuberculosis isoniazid-resistant isolates by PCR-RFLP. Materials and Methods: Polymorphism analysis of 1054 and 630 bp products containing the S315T mutation of the katG gene was performed by PCR-RFLP using the MspI and SatI restriction enzymes from 68 Mycobacterium tuberculosis isolates. Sensitivity, specificity, positive and negative predictive values, positive and negative likelihood ratio were determined using SPSS. Results: 74.46% of isoniazid-resistant and 4.76% of isoniazid-sensitive isolates showed the S315T mutation in katG gene. The PCR-RFLP for S315T of the katG gene had 85.4% sensitivity and 95.2% specificity with MspI and 85.4% sensitivity and 94.4% specificity with SatI. Discussion: The PCR-RFLP has a high resolutive capacity that depends on the enzyme that is used as it was observed in previous studies. The presence of the S315T mutation in treatment-naive patients suggests the circulation of isolates resistant to isoniazid. Conclusion: PCR-RFLP is a valid and rapid alternative for the diagnosis of isoniazid resistance, by detection of S315T mutation in the katG gene compared to the conventional method of proportions.

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“…Las mutaciones en los genes de Mycobacterium tuberculosis katG e inhA, han sido reportados como los responsables del 60 -80% de los aislados resistentes a isoniacida [27][28][29][30] .…”

Section: Discussionunclassified

Determinación de la mutación S315T del gen katG en aislados resistentes a Isoniacida de Mycobacterium tuberculosis mediante PCR-RFLP

Sánchez-Domínguez

Nicola-Salas

Morey-León

2018

Infect

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Section: Discussionmentioning

confidence: 99%

“…On the other hand, katG has been linked to resistance to SM, RIF, and EMB in addition to INH. 16 The ahpC gene of MTB encodes the antioxidant enzyme catalase-peroxidase that causes mutations in katG and is known to be associated with resistance to INH, an anti-TB drug. [12][13][14]17 However, ahpC-associated mutations were not detected in this study.…”

Section: Introductionmentioning

confidence: 99%

“…Of the 69 cases lacking INH resistance mutations in katG, 6 (8.7%) nonetheless showed INH resistance, suggesting that this phenotype is associated with genes other than katG.On the other hand, katG has been linked to resistance to SM, RIF, and EMB in addition to INH. 16 The ahpC gene of MTB encodes the antioxidant enzyme catalase-peroxidase that causes mutations in katG and is known to be associated with resistance to INH, an anti-TB drug. [12][13][14]17 However, ahpC-associated mutations were not detected in this study.Using the MGIT, Rigouts et al 18 reported that all strains harboring Asp516Val or Asp516Phe mutations in rpoB were resistant to RIF, but 1 of the 6 strains with the Asp516Tyr mutation showed RIF resistance, and only at 0.5 μg/mL.…”

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confidence: 99%

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Genetic and phenotypic characterizations of drug‐resistant Mycobacterium tuberculosis isolates in Cheonan, Korea

Jeon

Kim

Choi

et al. 2018

Clinical Laboratory Analysis

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“…The region of the 435 bp (669-1103bp) katG gene was amplified with a set of primers F-GCAGATGGGGCTGATCTACG and R-AACGGGTCCGGGATGG TG. Before sequencing, PCR amplification reactions were modified following (Farnia et al 2010) and (Jaiswal et al 2017). Final volumes of 50 µL were used for the rpoB and katG genes.…”

Section: Sample Sources and Processingmentioning

confidence: 99%

Novel mutations detected from drug resistant Mycobacterium tuberculosis isolated from North East of Thailand

Thwe

Namwat

Pinlaor

et al. 2021

World J Microbiol Biotechnol

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The emergence of drug-resistant tuberculosis is a major global public health threat. Thailand is one of the top 14 countries with high tuberculosis, tuberculosis co-infected with Human Immunodeficiency Virus (HIV) and multi-drug resistant tuberculosis rates. Immediate detection of drug-resistant tuberculosis is necessary to reduce mortality and morbidity by effectively providing treatment to ameliorate the formation of resistant strains. Limited data exist of mutation profiles in Northeastern Thailand. Here, 65 drug-resistant Mycobacterium tuberculosis isolates were used to detect mutations by polymerase chain reaction (PCR) and DNA sequencing. In the katG gene, mutations occurred in 47(79.7%) among 59 isoniazid resistant samples. For rpoB gene, 31 (96.9%) were observed as mutations in 32 rifampicin resistant isolates. Of 47 katG mutation samples, 45 (95.7%) had mutations in katG315 codon and 2 (4.3%) showed novel mutations at katG365 with amino acid substitution of CCG-CGG (Pro-Arg). Moreover, 18 (58.1%) mutations at rpoB531, 11 (35.5%) mutations at rpoB516, 10 (32.3%) mutations at rpoB526 and 1 (3.2%) mutation at rpoB533 were found between 31 rifampicin resistant samples. Common and novel mutations of the rpoB and katG genes in Northeastern Thailand were generated. DNA sequencing showed high accuracy, while conventional polymerase chain reaction could be applied as an initial marker for screening rifampicin and isoniazid drug-resistant Mycobacterium tuberculosis in Northeastern Thailand. Mutations reported here should be considered when developing new molecular diagnostic methods for implementation in Northeastern Thailand.

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Mutations in katG and inhA genes of isoniazid-resistant and -sensitive clinical isolates of Mycobacterium tuberculosis from cases of pulmonary tuberculosis and their association with minimum inhibitory concentration of isoniazid

Abstract: c l i n i c a l e p i d e m i o l o g y a n d g l o b a l h e a l t h 5 (2 0 1 7) 1 4 3-1 4 7

Cited by 7 publications

References 22 publications

Determinación de la mutación S315T del gen katG en aislados resistentes a Isoniacida de Mycobacterium tuberculosis mediante PCR-RFLP

Determinación de la mutación S315T del gen katG en aislados resistentes a Isoniacida de Mycobacterium tuberculosis mediante PCR-RFLP

Genetic and phenotypic characterizations of drug‐resistant Mycobacterium tuberculosis isolates in Cheonan, Korea

Novel mutations detected from drug resistant Mycobacterium tuberculosis isolated from North East of Thailand

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