Skip to main content
SpringerLink
Log in

Use of the INNO LiPA Rif.TB for detection of Mycobacterium tuberculosis DNA directly in clinical specimens and for simultaneous determination of rifampin susceptibility

  • Article
  • Published:
European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

The INNO LiPA Rif.TB (Innogenetics, Ghent, Belgium) is a reverse hybridization test developed to detect genetic markers of resistance to rifampin in Mycobacterium tuberculosis complex. In the present study, this test was used directly on 3,763 clinical specimens by adopting a nested amplification of the target. The specificity of the system (98.4%) was optimal, but sensitivity (69.5%) was unsatisfactory. However, when use of the system was limited to smear-positive specimens, the sensitivity rose to 91.7%. As expected, the ability of the system to predict rifampin resistance was not influenced by its direct use on clinical specimens and confirmed the favorable results repeatedly reported in the literature.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. World Health Organization (2005) Global tuberculosis programme. Anti-tuberculosis drug resistance in the world. The WHO/IUATLD global project on anti-tuberculosis drug resistance surveillance 1994–1997. WHO, Geneva, Switzerland, pp 1–153

    Google Scholar 

  2. Pfyffer GE, Brown-Elliott BA, Wallace RJ Jr (2003) Mycobacterium: general characteristics, isolation, and staining procedures. In: Murray PR, Baron EJ, Jorgensen JH, Pfaller MA, Yolken RH (eds) Manual of clinical microbiology. ASM Press, Washington DC, pp 532–559

    Google Scholar 

  3. Cooksey RC (2003) Recent advances in laboratory procedures for pathogenic mycobacteria. Clin Lab Med 23:801–821

    Article  PubMed  Google Scholar 

  4. Drobniewski FA, Caws M, Gibson A, Young D (2003) Modern laboratory diagnosis of tuberculosis. Lancet Infect Dis 3:141–147

    Article  PubMed  CAS  Google Scholar 

  5. Abe C, Ogata H, Kawata K, Hiraga T, Takashima T, Suetake T (2000) Detection of rifampin-resistant Mycobacterium tuberculosis by line probe assay (LiPA). Kekkaku 75:575–581

    PubMed  CAS  Google Scholar 

  6. Bártfai Z, Somoskövi A, Ködmön C, Szabó N, Puskás E, Kosztolányi L, Faragó E, Mester J, Parsons LM, Salfinger M (2001) Molecular characterization of rifampin-resistant isolates of Mycobacterium tuberculosis from Hungary by DNA sequencing and the line probe assay. J Clin Microbiol 39:3736–3739

    Article  PubMed  Google Scholar 

  7. Cooksey RC, Morlock GP, Glickman S, Crawford JT (1997) Evaluation of a line probe assay kit for characterization of rpoB mutations in rifampin-resistant Mycobacterium tuberculosis isolates from New York City. J Clin Microbiol 35:1281–1283

    PubMed  CAS  Google Scholar 

  8. Gamboa F, Cardona PJ, Manterola JM, Lonca J, Matas L, Padilla E, Manzano JR, Ausina V (1998) Evaluation of a commercial probe assay for detection of rifampin resistance in Mycobacterium tuberculosis directly from respiratory and nonrespiratory clinical samples. Eur J Clin Microbiol Infect Dis 17:189–192

    PubMed  CAS  Google Scholar 

  9. Hirano K, Abe C, Takahasci M (1999) Mutations of rpoB gene of rifampin-resistant Mycobacterium tuberculosis strains isolated mostly in Asian countries and their rapid detection by line probe assay. J Clin Microbiol 37:2663–2666

    PubMed  CAS  Google Scholar 

  10. Johansen IS, Lundgren B, Sosnovskaja A, Thomsen VØ (2003) Direct detection of multidrug-resistant Mycobacterium tuberculosis in clinical specimens in low- and high-incidence countries by line probe assay. J Clin Microbiol 41:4454–4456

    Article  PubMed  CAS  Google Scholar 

  11. Matsiota-Bernard P, Vrioni G, Marinis E (1998) Characterization of rpoB mutations in rifampin-resistant clinical Mycobacterium tuberculosis isolates from Grece. J Clin Microbiol 36:20–23

    PubMed  CAS  Google Scholar 

  12. Mokrousov I, Filliol I, Legrand E, Sola C, Otten T, Vyshnevskaya E, Limeschenko E, Vyshnevskiy B, Narvskaya O, Rastogi N (2002) Molecular characterization of multiple-drug-resistant Mycobacterium tuberculosis isolates from northwestern Russia and analysis of rifampin resistance using RNA/RNA mismatch analysis as compared to the line probe assay and sequencing of the rpoB gene. Res Microbiol 153:213–219

    Article  PubMed  CAS  Google Scholar 

  13. Watterson SA, Wilson SM, Yates MD, Drobniewski FA (1998) Comparison of three molecular assays for rapid detection of rifampin resistance in Mycobacterium tuberculosis. J Clin Microbiol 36:1969–1973

    PubMed  CAS  Google Scholar 

  14. Morgan M, Kalantri S, Flores L, Pai M (2005) A commercial line probe assay for the rapid detection of rifampicin resistance in Mycobacterium tuberculosis: a systematic review and meta-analysis. BMC Infect Dis 5:62

    Article  PubMed  Google Scholar 

  15. Tortoli E, Mariottini A, Mazzarelli G (2003) Evaluation of INNO-LiPA MYCOBACTERIA v2: improved reverse hybridization multiple DNA probe assay for mycobacterial identification. J Clin Microbiol 41:4418–4420

    Article  PubMed  CAS  Google Scholar 

  16. Richter E, Weizenegger M, Fahr AM, Rüsch-Gerdes S (2004) Usefulness of the GenoType MTBC assay for differentiating species of the Mycobacterium tuberculosis complex in cultures obtained from clinical specimens. J Clin Microbiol 42:4303–4306

    Article  PubMed  CAS  Google Scholar 

  17. Cloud JL, Neal H, Rosenberry R, Turenne CY, Jama M, Hillyard DR, Carroll KC (2002) Identification of Mycobacterium spp. by using a commercial 16S ribosomal DNA sequencing kit and additional sequencing libraries. J Clin Microbiol 40:400–406

    Article  PubMed  CAS  Google Scholar 

  18. Siddiqi SH, Libonati JP, Middlebrook G (1981) Evaluation of a rapid radiometric method for drug susceptibility testing of Mycobacterium tuberculosis. J Clin Microbiol 13:908–912

    PubMed  CAS  Google Scholar 

  19. Davidoff F (1999) Standing statistic right side up. Ann Intern Med 130:1019–1021

    PubMed  CAS  Google Scholar 

  20. Heifets LB, Cangelosi GA (1999) Drug susceptibility testing of Mycobacterium tuberculosis: a neglected problem at the turn of the century. Int J Tuberc Lung Dis 3:564–581

    PubMed  CAS  Google Scholar 

  21. World Health Organization (1998) Guidelines for surveillance of drug resistance in tuberculosis. Int J Tuberc Lung Dis 2:72–89

    Google Scholar 

  22. Cavusoglu C, Hilmioglu S, Guneri S, Bilgic A (2002) Characterization of rpoB mutations in rifampin-resistant clinical isolates of Mycobacterium tuberculosis from Turkey by DNA sequencing and line probe assay. J Clin Microbiol 40:4435–4438

    Article  PubMed  CAS  Google Scholar 

  23. De Beenhouwer H, Lhiang Z, Jannes G, Mijs W, Machtelinckx L, Rossau R, Traore H, 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 76:425–430

    Article  PubMed  Google Scholar 

  24. Higuchi T, Fushiwaki T, Tanaka N, Miyake S, Ogura T, Yoshida H, Takashima T, Nakagawa M, Maekura R, Hiraga T, Suetake T (2004) Direct detection of rifampicin resistant Mycobacterium tuberculosis in sputum by line probe assay (LiPA). Kekkaku 79:525–530 (in Japanese)

    PubMed  CAS  Google Scholar 

  25. Viveiros M, Leandro C, Rodrigues L, Almeida J, Bettencourt R, Couto I, Carrilho L, Diogo J, Fonseca A, Lito L, Lopes J, Pacheco T, Pessanha M, Quirim J, Sancho L, Salfinger M, Amaral L (2005) Direct application of the INNO-LiPA Rif.TB line-probe assay for rapid identification of Mycobacterium tuberculosis complex strains and detection of rifampin resistance in 360 smear-positive respiratory specimens from an area of high incidence of multidrug-resistant tuberculosis. J Clin Microbiol 43:4880–4884

    Article  PubMed  CAS  Google Scholar 

  26. Piersimoni C, Scarparo C (2003) Relevance of commercial amplification methods for direct detection of Mycobacterium tuberculosis complex in clinical samples. J Clin Microbiol 41:5355–5365

    Article  PubMed  CAS  Google Scholar 

  27. Bergmann JS, Woods GL (1996) Clinical evaluation of the Roche AMPLICOR PCR Mycobacterium tuberculosis test for detection of M. tuberculosis in respiratory specimens. J Clin Microbiol 34:1083–1085

    PubMed  CAS  Google Scholar 

  28. Bergmann JS, Keating WE, Woods GL (2000) Clinical evaluation of the BDProbeTec ET system for rapid detection of Mycobacterium tuberculosis. J Clin Microbiol 38:863–865

    PubMed  CAS  Google Scholar 

  29. Reischl U, Lehn N, Wolf H, Naumann L (1998) Clinical evaluation of the automated COBAS AMPLICOR MTB assay for testing respiratory and nonrespiratory specimens. J Clin Microbiol 36:2853–2860

    PubMed  CAS  Google Scholar 

  30. Wobeser WL, Krajden M, Conly J, Simpson H, Yim B, D’Costa M, Fuksa M, Hian-Cheong C, Petterson M, Phillips A, Bannatyne R, Haddad A, Brunton JL, Krajden S (1996) Evaluation of Roche Amplicor PCR assay for Mycobacterium tuberculosis. J Clin Microbiol 34:134–139

    PubMed  CAS  Google Scholar 

  31. Hillemann D, Weizenegger M, Kubica T, Richter E, Niemann S (2005) Use of the GenoType MTBDR assay for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis complex isolates. J Clin Microbiol 43:3699–3703

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

We are indebted to M. Coyle who carefully reviewed the manuscript. We thank G. Cecchi and M. Morrisi for technical work.

Author information

Authors and Affiliations

  1. Regional Reference Center for Mycobacteria, Microbiology Laboratory, Careggi Hospital, viale Morgagni 85, 50134, Florence, Italy

    E. Tortoli & F. Marcelli

Authors
  1. E. Tortoli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F. Marcelli
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to E. Tortoli.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tortoli, E., Marcelli, F. Use of the INNO LiPA Rif.TB for detection of Mycobacterium tuberculosis DNA directly in clinical specimens and for simultaneous determination of rifampin susceptibility. Eur J Clin Microbiol Infect Dis 26, 51–55 (2007). https://doi.org/10.1007/s10096-006-0240-x

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10096-006-0240-x

Keywords

Search

Navigation