Skip to main content
The Egyptian Journal of Bronchology
Download PDF
  • Original article
  • Open access
  • Published:

Real-time polymerase chain reaction as an alternative method for diagnosis of multidrug-resistant tuberculosis: can it stand alone in this concern

Egyptian Journal of Bronchology volume 11, pages 342–345 (2017)Cite this article

Abstract

Background

Delays in diagnosing multidrug-resistant tuberculosis (MDR-TB) are responsible for higher tuberculosis morbidity and mortality and its subsequent transmission. Molecular assays such as real-time PCR (RT-PCR) used to identify drug resistance in Mycobacterium tuberculosis are more rapid than standard drug susceptibility testing.

Objectives

The aim of this study was to evaluate the diagnostic performance of the Anyplex MTB/MDR RT-PCR assay in detecting MDR-TB strains.

Patients and methods

Sputum samples were collected from 29 patients with symptoms and radiological findings suggestive of active pulmonary tuberculosis, with at least one of three sputum smear samples showing acid-fast bacilli and/ or sputum culture isolates positive for M. tuberculosis. The results obtained by RT-PCR were compared with those obtained by the Mycobacterium growth indicator tube SIRE method.

Results

M. tuberculosis was confirmed in 29 specimens. Only six cases determined as MDR-TB were obtained by Mycobacterium growth indicator tube SIRE. For detection of rifampicin-resistant and isoniazid-resistant strains, the RT-PCR assay yielded a sensitivity of 62.5 and 66.66% and specificity of 80 and 95%, respectively. The overall sensitivity of that assay was 64.2% and specificity was 88.88%.

Conclusion

RT-PCR is an easy and reliable assay for rapid detection of MDR-TB in clinical specimens. However, RT-PCR should be followed by a culture method to increase the overall sensitivity of that assay.

References

  1. Espinal MA. The global situation of MDR-TB. Tuberculosis (Edinb) 2003; 83:44–51.

    Article  Google Scholar 

  2. Soini H, Musser JM. Molecular diagnosis of mycobacteria. Clin Chem 2001; 47:809–814.

    Article  CAS  Google Scholar 

  3. Lee AS, Lim IH, Tang LL, Wong SY. High frequency of mutations in the rpoB gene in rifampin-resistant clinical isolates of Mycobacterium tuberculosis from Singapore. J Clin Microbiol 2005; 43:2026–2027.

    Article  CAS  Google Scholar 

  4. Hazbon MH, Brimacombe M, Bobadilla del Valle M, Cavatore M, Guerrero MI, Varma-Basil M, et al. Population genetics study of isoniazid resistance mutations and evolution of multidrug-resistant Mycobacterium tuberculosis. Antimicrob Agents Chemother 2006; 50:2640–2649.

    Article  CAS  Google Scholar 

  5. Bergval IL, Vijzelaar RN, Dalla Costa ER, Schuitema AR, Oskam L, Kritski AL, et al. Development of multiplex assay for rapid characterization of Mycobacterium tuberculosis. J Clin Microbiol 2008; 46:689–699.

    Article  CAS  Google Scholar 

  6. Ong DC, Yam WC, Siu GK, Lee AS. Rapid detection of rifampicin- and isoniazid-resistant Mycobacterium tuberculosis by high-resolution melting analysis. J Clin Microbiol 2010; 48:1047–1054.

    Article  CAS  Google Scholar 

  7. Lagrange PH, Thangaraj SK, Dayal R, Despande A, Ganguly NK, Girardi E, et al. A toolbox for tuberculosis diagnosis: an Indian multicentric study (2006-2008): microbiological results. PloS one 2012; 7:e43739.

  8. Kent PT, Kubica GP. Public Health mycobacteriology: A guide for the level III laboratory. Georgia: US Departement of Health and Human Services, CDC Atlanta; 1985.

  9. Barenfanger J. Making your lab safe against multi-drug resistant Mycobacterium tuberculosis. Clin Microbiol Newsl 1993; 15:76–80.

    Article  Google Scholar 

  10. Greer CE, Wheeler CM, Manos MM. Sample preparation and PCR amplification from paraffin-embedded tissues. PCR Methods Appl 1994; 3:S113–S122.

    Article  CAS  Google Scholar 

  11. ElFeky DS, ElShimy A. Evaluation of the diagnostic performance of Anyplex II MTB/MDR/XDR detection assay for Detection and antimicrobial susceptibility testing of Mycobacterium tuberculosis from sputum specimens. Int J Curr Microbiol App Sci 2015; 4:657–669.

    CAS  Google Scholar 

  12. Ramirez MV, Cowart KC, Campbell PJ, Morlock GP, Sikes D, Winchell JM, et al. Rapid detection of multidrug-resistant Mycobacterium tuberculosis by use of real-time PCR and high-resolution melt analysis. J Clin Microbiol 2010; 48:4003–4009.

    Article  CAS  Google Scholar 

  13. Goncalves MG, Fukasawa LO, Oliveira RS, Salgado MM, Harrison LH, Shutt KA, et al. Fast test for assessing the susceptibility of Mycobacterium tuberculosis to isoniazid and rifampin by real-time PCR. Mem Inst Oswaldo Cruz 2012; 107:903–908.

    Article  CAS  Google Scholar 

  14. El-Sayed Zaki M, Hassanin N. DNA sequence analysis of rpoB Gene Mutations in rifampicin resistant Mycobacterium tuberculosis. J Rapid Meth Aut Microbiol 2006; 14:237–248.

    Article  Google Scholar 

  15. Abdullal AK. Direct and rapid detection of multidrug resistant Mycobacterium tuberculosis (MDR-MT) in sputum samples. London, UK: World Congress of Infectious Diseases 10–12 August 2015.

  16. Chan RC, Hui M, Chan EW, Au TK, Chin ML, Yip CK, et al. Genetic and phenotypic characterization of drug-resistant Mycobacterium tuberculosis isolates in Hong Kong. J Antimicrob Chemother 2007; 59:866–873.

    Article  CAS  Google Scholar 

  17. Nimri L, Samara H, Batchoun R. Detection of mutations assaciated with multidrug resistant Mycobacterium tuberculosis clinical isolates. FEMS Immunol Med Microbiol 2011; 62:321–327.

    Article  CAS  Google Scholar 

  18. Plinke C, Cox HS, Kalon S, Doshetov D, Rusch-Gerdes S, Niemann S. Tuberculosis ethambutol resistance: concordance between phenotypic and genotypic test results. Tuberculosis (Edinb) 2009; 89:448–452.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. ERS National Delegate of Egypt, Assuit University, Assuit, Egypt

    Gamal Rabie Agamy

  2. Fellow at Chest Diseases and Tuberculosis Department, Faculty of Medicine, Assuit University Hospital, Assuit, 71526, Egypt

    Gamal Rabie Agamy, Safaa Moktar Wafy & Lamees Mohamed Bakar PhD

  3. Microbiology and Clinical Pathology Department, Faculty of Medicine, Assuit University, Assuit, Egypt

    Asmaa Omar Ahmed

Authors
  1. Gamal Rabie Agamy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Safaa Moktar Wafy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Asmaa Omar Ahmed
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lamees Mohamed Bakar PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lamees Mohamed Bakar PhD.

Additional information

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work noncommercially, as long as the author is credited and the new creations are licensed under the identical terms.

Rights and permissions

This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Agamy, G.R., Wafy, S.M., Ahmed, A.O. et al. Real-time polymerase chain reaction as an alternative method for diagnosis of multidrug-resistant tuberculosis: can it stand alone in this concern. Egypt J Bronchol 11, 342–345 (2017). https://doi.org/10.4103/1687-8426.217631

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.4103/1687-8426.217631

Keywords