Comparison of polymerase chain reaction single-strand conformation polymorphism with DNA sequencing to detect drug resistance of mycobacterium tuberculosis isolates

Maryam Fatemeh Sheikholslami; Parissa Farnia; Payam Tabarsi; Mouayad Aghali Merza

Archives of Clinical Infectious Diseases

Infectious Diseases and Tropical Medicine Research Center, SBUMS

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Comparison of polymerase chain reaction single-strand conformation polymorphism with DNA sequencing to detect drug resistance of mycobacterium tuberculosis isolates

Author(s):

Maryam Fatemeh Sheikholslami^1,*,

Parissa Farnia¹,

Payam Tabarsi¹,

Mouayad Aghali Merza¹

1Mycobacteriology Research Center

Archives of Clinical Infectious Diseases:Vol. 6, issue 2; e93411

Published online:Apr 30, 2011

Article type:Research Article

Received:May 12, 2019

Accepted:Feb 07, 2011

How to Cite:Maryam Fatemeh SheikholslamiParissa FarniaPayam TabarsiMouayad Aghali MerzaComparison of polymerase chain reaction single-strand conformation polymorphism with DNA sequencing to detect drug resistance of mycobacterium tuberculosis isolates.Arch Clin Infect Dis.6(2):e93411.

Abstract

Objectives: Advancements in molecular technology increased our understanding of genetic mechanism of drug resistance. Nowadays, the chance of rapid detection of resistant Mycobacterium tuberculosis (M. tuberculosis) strains is increased. In the present study, we aimed to investigate the sensitivity and specificity of PCR-SSCP for detecting susceptible and resistant strains of M. tuberculosis compared with DNA sequencing.

To calculate the sensitivity and specificity of PCR-SSCP assay to detect drug resistance in M. tuberculosis, respiratory samples were collected from suspected patients referred to Mycobacteriology Research Center (Masieh Daneshvary Hosptial) since 2002. Susceptibility testing against first line drugs was performed on 74 culturepositive specimens. Consequently, PCR-SSCP and DNA sequencing were performed on katG, inhA, ahpC and rpoB genes.

Results: Drug-susceptibility testing by the proportional method in selected samples revealed 16 MDR (21.6%), 23 mono-drug resistant (31%) and 35 susceptible strains (47.3%). In comparison with DNA sequencing as a gold standard for molecular methods, the sensitivity of PCR-SSCP assay for detecting of mutation in 315 codon of katG gene was 94.74% (CI=73.97%-99.87%) with 100% (CI=93.51%-100%) specificity. In contrast, the sensitivity and specificity of this assay in detecting of rpoB gene were 70.8 (CI=48.91%-87.38%) and 88 (CI=75.69%-95.47%), respectively.

Conclusion: PCR-SSCP in combination with DNA sequencing can be used as screening method to detect MDR-TB and mono-drug resistant cases.

Keywords

polymerase chain reaction

Mycobacterium tuberculosis

drug resistance

References

1.
The references are available in PDF file.

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