Genotypic detection of fluoroquinolone resistance in drug-resistant Mycobacterium tuberculosis at a tertiary care centre in south Coastal Karnataka, India

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Abstract

Objectives: This study aimed to characterise mutations associated with fluoroquinolone resistance in drug-resistant Mycobacterium tuberculosis clinical isolates at a tertiary care centre in south Coastal Karnataka, India. Methods: DNA from 50 M. tuberculosis clinical isolates was extracted and the gyrA and gyrB genes were amplified. Purified amplicons of gyrA and gyrB were sequenced and extended-sequencing PCR products were analysed. Analysis of mutations in gyrA and gyrB was done using the MUBII-TB-DB database. Statistical analysis was performed using SPSS v.22 and data were compared by χ2 test. A P-value of <0.05 was considered statistically significant. Results: Mutations conferring resistance to fluoroquinolones were observed in 9 isolates (18%). The gyrA A281G (D94G) mutation was observed in 3 isolates (6%), whereas mutations G280T (D94Y) and A281C (D94A) were each observed in 1 isolate (2%). Mutation G1498A (D500N) in gyrB alone was observed in 2 isolates (4%). Two isolates (4%) had mutations both in gyrA and gyrB; gyrA mutation T271C (S91P) was observed in one isolate, whereas the other isolate had gyrA C269G (A90G), but both isolates had a common G1498A (D500N) gyrB mutation. G284C mutation conferring S95T polymorphism (not associated with fluoroquinolone resistance) was observed in 39/50 isolates (78%). Conclusion: gyrA mutations at codons 94, 91 and 90 and gyrB mutation G1498A (D500N) were the most common mutations associated with fluoroquinolone resistance in clinical isolates in this study. Future studies including a larger number of samples are desirable to fully explore the true extent of fluoroquinolone resistance and mutations associated with them.

Original languageEnglish
Pages (from-to)250-253
Number of pages4
JournalJournal of Global Antimicrobial Resistance
Volume13
DOIs
Publication statusPublished - 01-06-2018

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Multidrug-Resistant Tuberculosis
Fluoroquinolones
Mycobacterium tuberculosis
Tertiary Care Centers
India
Mutation
Codon

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology and Allergy
  • Immunology
  • Microbiology (medical)

Cite this

@article{1dd87db3006e493fa79bf3f7beff8c0f,
title = "Genotypic detection of fluoroquinolone resistance in drug-resistant Mycobacterium tuberculosis at a tertiary care centre in south Coastal Karnataka, India",
abstract = "Objectives: This study aimed to characterise mutations associated with fluoroquinolone resistance in drug-resistant Mycobacterium tuberculosis clinical isolates at a tertiary care centre in south Coastal Karnataka, India. Methods: DNA from 50 M. tuberculosis clinical isolates was extracted and the gyrA and gyrB genes were amplified. Purified amplicons of gyrA and gyrB were sequenced and extended-sequencing PCR products were analysed. Analysis of mutations in gyrA and gyrB was done using the MUBII-TB-DB database. Statistical analysis was performed using SPSS v.22 and data were compared by χ2 test. A P-value of <0.05 was considered statistically significant. Results: Mutations conferring resistance to fluoroquinolones were observed in 9 isolates (18{\%}). The gyrA A281G (D94G) mutation was observed in 3 isolates (6{\%}), whereas mutations G280T (D94Y) and A281C (D94A) were each observed in 1 isolate (2{\%}). Mutation G1498A (D500N) in gyrB alone was observed in 2 isolates (4{\%}). Two isolates (4{\%}) had mutations both in gyrA and gyrB; gyrA mutation T271C (S91P) was observed in one isolate, whereas the other isolate had gyrA C269G (A90G), but both isolates had a common G1498A (D500N) gyrB mutation. G284C mutation conferring S95T polymorphism (not associated with fluoroquinolone resistance) was observed in 39/50 isolates (78{\%}). Conclusion: gyrA mutations at codons 94, 91 and 90 and gyrB mutation G1498A (D500N) were the most common mutations associated with fluoroquinolone resistance in clinical isolates in this study. Future studies including a larger number of samples are desirable to fully explore the true extent of fluoroquinolone resistance and mutations associated with them.",
author = "Kiran Chawla and Ajay Kumar and Shenoy, {Vishnu Prasad} and Sanjiban Chakrabarty and Kapaettu Satyamoorthy",
year = "2018",
month = "6",
day = "1",
doi = "10.1016/j.jgar.2018.01.023",
language = "English",
volume = "13",
pages = "250--253",
journal = "Journal of Global Antimicrobial Resistance",
issn = "2213-7165",
publisher = "Elsevier BV",

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TY - JOUR

T1 - Genotypic detection of fluoroquinolone resistance in drug-resistant Mycobacterium tuberculosis at a tertiary care centre in south Coastal Karnataka, India

AU - Chawla, Kiran

AU - Kumar, Ajay

AU - Shenoy, Vishnu Prasad

AU - Chakrabarty, Sanjiban

AU - Satyamoorthy, Kapaettu

PY - 2018/6/1

Y1 - 2018/6/1

N2 - Objectives: This study aimed to characterise mutations associated with fluoroquinolone resistance in drug-resistant Mycobacterium tuberculosis clinical isolates at a tertiary care centre in south Coastal Karnataka, India. Methods: DNA from 50 M. tuberculosis clinical isolates was extracted and the gyrA and gyrB genes were amplified. Purified amplicons of gyrA and gyrB were sequenced and extended-sequencing PCR products were analysed. Analysis of mutations in gyrA and gyrB was done using the MUBII-TB-DB database. Statistical analysis was performed using SPSS v.22 and data were compared by χ2 test. A P-value of <0.05 was considered statistically significant. Results: Mutations conferring resistance to fluoroquinolones were observed in 9 isolates (18%). The gyrA A281G (D94G) mutation was observed in 3 isolates (6%), whereas mutations G280T (D94Y) and A281C (D94A) were each observed in 1 isolate (2%). Mutation G1498A (D500N) in gyrB alone was observed in 2 isolates (4%). Two isolates (4%) had mutations both in gyrA and gyrB; gyrA mutation T271C (S91P) was observed in one isolate, whereas the other isolate had gyrA C269G (A90G), but both isolates had a common G1498A (D500N) gyrB mutation. G284C mutation conferring S95T polymorphism (not associated with fluoroquinolone resistance) was observed in 39/50 isolates (78%). Conclusion: gyrA mutations at codons 94, 91 and 90 and gyrB mutation G1498A (D500N) were the most common mutations associated with fluoroquinolone resistance in clinical isolates in this study. Future studies including a larger number of samples are desirable to fully explore the true extent of fluoroquinolone resistance and mutations associated with them.

AB - Objectives: This study aimed to characterise mutations associated with fluoroquinolone resistance in drug-resistant Mycobacterium tuberculosis clinical isolates at a tertiary care centre in south Coastal Karnataka, India. Methods: DNA from 50 M. tuberculosis clinical isolates was extracted and the gyrA and gyrB genes were amplified. Purified amplicons of gyrA and gyrB were sequenced and extended-sequencing PCR products were analysed. Analysis of mutations in gyrA and gyrB was done using the MUBII-TB-DB database. Statistical analysis was performed using SPSS v.22 and data were compared by χ2 test. A P-value of <0.05 was considered statistically significant. Results: Mutations conferring resistance to fluoroquinolones were observed in 9 isolates (18%). The gyrA A281G (D94G) mutation was observed in 3 isolates (6%), whereas mutations G280T (D94Y) and A281C (D94A) were each observed in 1 isolate (2%). Mutation G1498A (D500N) in gyrB alone was observed in 2 isolates (4%). Two isolates (4%) had mutations both in gyrA and gyrB; gyrA mutation T271C (S91P) was observed in one isolate, whereas the other isolate had gyrA C269G (A90G), but both isolates had a common G1498A (D500N) gyrB mutation. G284C mutation conferring S95T polymorphism (not associated with fluoroquinolone resistance) was observed in 39/50 isolates (78%). Conclusion: gyrA mutations at codons 94, 91 and 90 and gyrB mutation G1498A (D500N) were the most common mutations associated with fluoroquinolone resistance in clinical isolates in this study. Future studies including a larger number of samples are desirable to fully explore the true extent of fluoroquinolone resistance and mutations associated with them.

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U2 - 10.1016/j.jgar.2018.01.023

DO - 10.1016/j.jgar.2018.01.023

M3 - Article

VL - 13

SP - 250

EP - 253

JO - Journal of Global Antimicrobial Resistance

JF - Journal of Global Antimicrobial Resistance

SN - 2213-7165

ER -