Skip to main content
SpringerLink
Log in

Validation of in House PCR Using IS6110 for Detection of M. tuberculosis and Its Comparison with ZN Staining, Cultures and RT PCR Kit Methods

  • Conference paper
  • First Online:
Techno-Societal 2016 (ICATSA 2016)

  • 1146 Accesses

Abstract

A polymerase chain reaction (PCR) assay for the rapid detection of Mycobacterium tuberculosis in sputum samples was studied. The target DNA of 123 base pair bp fragment of IS6110 which was repeated in the M. tuberculosis genome and was highly specific for the M. tuberculosis complex. PCR was positive in all 51 smear and culture-positive samples, in 17 of 17 smear-negative and culture-positive samples, 8 of 132 smear and culture negative samples. The overall sensitivity and specificity were 84 and 94%. Rapid detection of MTB using commercial diagnostic systems is limited by their costs and/or their requirement of specific equipment. The Genosens’s MTB Complex/MOTT Qualitative Real Time PCR assay kit for direct detection of M. tuberculosis complex in pulmonary tuberculosis samples have been used in MGM CRL on routine basis and also compatible with Light Cycle 480 Real time PCR Instrument. Conventional polymerase chain reaction has a much higher sensitivity than microscopy and can facilitate therapeutic decisions for those with suspected pulmonary tuberculosis. Thus, IS6110 as a PCR target was found to be very useful for rapid diagnosis of M. tuberculosis infection and start of antituberculous chemotherapy.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 259.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 329.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 329.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Jaggarajamma K, Sudha G, Chandrasekaran V, Nirupa C, Thomas A, Santha T, Narayanan PR (2007) Reasons for non-compliance among patients treated under Revised National Tuberculosis Control Programme (RNTCP), Tiruvallur district, South India. Ind J Tub 54(3):130

    Google Scholar 

  2. Aslanzadeh J, De la Viuda M, Fille M, Smith WB, Namdari H (1998) Comparison of culture and acid-fast bacilli stain to PCR for detection of Mycobacterium tuberculosisin clinical samples. Mol cell probes 12(4):207–211

    Google Scholar 

  3. Nolte FS, Metchock B, McGowan JE, Edwards A, Okwumabua O, Thurmond C, Shinnick T (1993) Direct detection of Mycobacterium tuberculosis in sputum by polymerase chain reaction and DNA hybridization. J Clin Microbiol 31(7):1777–1782

    Google Scholar 

  4. Kocagöz T, Yilmaz E, Ozkara S, Kocagöz S, Hayran M, Sachedeva M, Chambers HF (1993) Detection of Mycobacterium tuberculosis in sputum samples by polymerase chain reaction using a simplified procedure. J Clin Microbiol 31(6):1435–1438

    Google Scholar 

  5. García-Quintanilla A, Garcia L, Tudó G, Navarro M, González J, de Anta MTJ (2000) Single-tube balanced heminested PCR for detecting Mycobacterium tuberculosis in smear-negative samples. J Clin Microbiol 38(3):1166–1169

    Google Scholar 

  6. Gengvinij N, Pattanakitsakul SN, Chierakul N, Chaiprasert A (2001) Detection of Mycobacterium tuberculosis from sputum specimens using one-tube nested PCR

    Google Scholar 

  7. Lemaître N, Armand S, Vachée A, Capilliez O, Dumoulin C, Courcol RJ (2004) Comparison of the real-time PCR method and the Gen-Probe amplified Mycobacterium tuberculosis direct test for detection of Mycobacterium tuberculosis in pulmonary and nonpulmonary specimens. J Clin Microbiol 42(9):4307–4309

    Article  Google Scholar 

  8. Takahashi T, Nakayama T (2006) Novel technique of quantitative nested real-time PCR assay for Mycobacterium tuberculosis DNA. J Clin Microbiol 44(3):1029–1039

    Article  Google Scholar 

  9. De Beenhouwer H, Lhiang Z, Jannes GA, Mijs W, Machtelinckx L, Rossau R, Portaels F (1995) Rapid detection of rifampicin resistance in sputum and biopsy specimens from tuberculosis patients by PCR and line probe assay. Tuber Lung Dis 76(5):425–430

    Article  Google Scholar 

  10. Della LP (2004) Mycobacteriology and mycobacterial susceptibility tests. In Henry D (ed) Clinical microbiology procedures handbook, 2 edn. ASM Press

    Google Scholar 

  11. Kim SJ (2005) Drug-susceptibility testing in tuberculosis: methods and reliability of results. Eur Respir J 25(3):564–569

    Article  Google Scholar 

  12. Negi SS, Anand R, Pasha ST, Gupta S, Basir SF, Khare S, Lal S (2007) Diagnostic potential of IS6110, 38 kDa, 65 kDa and 85B sequence-based polymerase chain reaction in the diagnosis of Mycobacterium tuberculosis in clinical samples. Indian J Med Microbiol 25(1):43

    Article  Google Scholar 

  13. Shawar RM, El-Zaatari FA, Nataraj A, Clarridge JE (1993) Detection of Mycobacterium tuberculosis in clinical samples by two-step polymerase chain reaction and nonisotopic hybridization methods. J Clin Microbiol 31(1):61–65

    Google Scholar 

  14. Eisenach KD, Sifford MD, Cave MD, Bates JH, Crawford JT (1991) Detection of Mycobacterium tuberculosis in sputum samples using a polymerase chain reaction. Am Rev Respir Dis 144(5):1160–1163

    Article  Google Scholar 

  15. Negi SS, Khan SF, Gupta S, Pasha ST, Khare S, Lal S (2005) Comparison of the conventional diagnostic modalities, bactec culture and polymerase chain reaction test for diagnosis of tuberculosis. Indian J Med Microbiol 23(1):29

    Article  Google Scholar 

  16. Banavaliker JN, Bhalotra B, Sharma DC, Goel MK, Khandekar PS, Bose M (1998) Identification of Mycobacterium tuberculosis by polymerase chain reaction in clinical specimens. Indian J Tuberc 45:15–18

    Google Scholar 

  17. Bechnoosh A, Lieberman JM, Duke MB (1997) Stutman comparison of quantitative polymerase chain reaction, therapy for pulmonary tuberculosis. Diag Microb Infect Dis 29(2):9–73

    Google Scholar 

  18. D’Amato RF, Hochstein LH, Colaninno PM, Scardamaglia M, Kim K, Mastellone AJ, Miller A (1996) Application of the Roche Amplicor, Mycobacterium tuberculosis (PCR) test to specimens other than respiratory secretions. Diagn Microbiol Infect Dis 24(1):15–17

    Article  Google Scholar 

  19. Singh KK, Muralidhar M, Kumar A, Chattopadhyaya TK, Kapila K, Singh MK, Tyagi JS (2000) Comparison of in house polymerase chain reaction with conventional techniques for the detection of Mycobacterium tuberculosis DNA in granulomatous lymphadenopathy. J Clin Pathol 53(5):355–361

    Article  Google Scholar 

  20. Rimek D, Tyagi S, Kappe R (2002) Performance of an IS6110-based PCR assay and the COBAS AMPLICOR MTB PCR system for detection of Mycobacterium tuberculosis complex DNA in human lymph node samples. J Clin Microbiol 40(8):3089–3092

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biotechnology, MGM Institute of Health Science & MGM College of Engineering & Technology, Kamothe, Navi Mumbai, 410209, India

    Girish Pai & Mansee Thakur

  2. Department of Medical Microbiology, MGM Institute of Health Sciences, Kamothe, Navi Mumbai, 410209, India

    Harapriya Kar

  3. Department of Biotechnology & Genetics, MGM Institute of Health Sciences, Kamothe, Navi Mumbai, 410209, India

    D. S. Joshi

Authors
  1. Girish Pai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mansee Thakur
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Harapriya Kar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D. S. Joshi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Girish Pai .

Editor information

Editors and Affiliations

  1. SVERI's College of Engineering , Pandharpur, Maharashtra, India

    Prashant M. Pawar

  2. SVERI's College of Engineering , Pandharpur, Maharashtra, India

    Babruvahan P. Ronge

  3. Bhabha Atomic Research Centre , Mumbai, India

    R. Balasubramaniam

  4. SVERI's College of Engineering , Pandharpur, Maharashtra, India

    Sridevi Seshabhattar

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer International Publishing AG

About this paper

Cite this paper

Pai, G., Thakur, M., Kar, H., Joshi, D.S. (2018). Validation of in House PCR Using IS6110 for Detection of M. tuberculosis and Its Comparison with ZN Staining, Cultures and RT PCR Kit Methods. In: Pawar, P., Ronge, B., Balasubramaniam, R., Seshabhattar, S. (eds) Techno-Societal 2016. ICATSA 2016. Springer, Cham. https://doi.org/10.1007/978-3-319-53556-2_61

Download citation

  • DOI: https://doi.org/10.1007/978-3-319-53556-2_61

  • Published:

  • Publisher Name: Springer, Cham

  • Print ISBN: 978-3-319-53555-5

  • Online ISBN: 978-3-319-53556-2

  • eBook Packages: EngineeringEngineering (R0)

Publish with us

Policies and ethics

Search

Navigation