Advertisement

Fluoromycobacteriophages for Drug Susceptibility Testing (DST) of Mycobacteria

  • Mariana PiuriEmail author
  • Graham F. Hatfull
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1898)

Abstract

Fluoromycobacteriophages are a new class of reporter phages that contain Laboratorio fluorescent reporter genes (gfp, ZsYellow, and mCherry) and provide a simple means of revealing the metabolic state of mycobacterial cells and therefore their response to antibiotics. Here we described a simple and rapid method for drug susceptibility testing (DST) of Mycobacterium spp using a fluorescence microscope, a flow cytometer, or a fluorimeter in a convenient multiwell format.

Key words

Reporter phage Mycobacteria DST Fluoromycobacteriophage Antibiotics Resistance 

References

  1. 1.
    WHO (2014) Global Tuberculosis Report. World Health Organization, GenevaGoogle Scholar
  2. 2.
    Zumla A et al (2012) Drug-resistant tuberculosis--current dilemmas, unanswered questions, challenges, and priority needs. J Infect Dis 205(Suppl 2):S228–S240CrossRefGoogle Scholar
  3. 3.
    Watterson SA, Drobniewski FA (2000) Modern laboratory diagnosis of mycobacterial infections. J Clin Pathol 53(10):727–732CrossRefGoogle Scholar
  4. 4.
    WHO (2011) Automated real-time nucleic acid amplification technology for rapid and simultaneous detection of tuberculosis and rifampicin resistance. Xpert MTB/RIF system, GenevaGoogle Scholar
  5. 5.
    Rachow A et al (2011) Rapid and accurate detection of Mycobacterium tuberculosis in sputum samples by Cepheid Xpert MTB/RIF assay--a clinical validation study. PLoS One 6(6):e20458CrossRefGoogle Scholar
  6. 6.
    Bowles EC et al (2011) Xpert MTB/RIF(R), a novel automated polymerase chain reaction-based tool for the diagnosis of tuberculosis. Int J Tuberc Lung Dis 15(7):988–989CrossRefGoogle Scholar
  7. 7.
    Piuri M, Jacobs WR Jr, Hatfull GF (2009) Fluoromycobacteriophages for rapid, specific, and sensitive antibiotic susceptibility testing of Mycobacterium tuberculosis. PLoS One 4(3):e4870CrossRefGoogle Scholar
  8. 8.
    Piuri M et al (2013) Generation of affinity-tagged fluoromycobacteriophages by mixed assembly of phage capsids. Appl Environ Microbiol 79(18):5608–5615CrossRefGoogle Scholar
  9. 9.
    Wilson SM et al (1997) Evaluation of a new rapid bacteriophage-based method for the drug susceptibility testing of Mycobacterium tuberculosis. Nat Med 3(4):465–468CrossRefGoogle Scholar
  10. 10.
    Watterson SA et al (1998) Comparison of three molecular assays for rapid detection of rifampin resistance in Mycobacterium tuberculosis. J Clin Microbiol 36(7):1969–1973PubMedPubMedCentralGoogle Scholar
  11. 11.
    Albert H et al (2001) Evaluation of FASTPlaqueTB-RIF, a rapid, manual test for the determination of rifampicin resistance from Mycobacterium tuberculosis cultures. Int J Tuberc Lung Dis 5(10):906–911PubMedGoogle Scholar
  12. 12.
    Jacobs WR Jr et al (1993) Rapid assessment of drug susceptibilities of Mycobacterium tuberculosis by means of luciferase reporter phages. Science 260(5109):819–822CrossRefGoogle Scholar
  13. 13.
    Banaiee N et al (2003) Rapid identification and susceptibility testing of Mycobacterium tuberculosis from MGIT cultures with luciferase reporter mycobacteriophages. J Med Microbiol 52.(Pt 7:557–561CrossRefGoogle Scholar
  14. 14.
    Marais BJ et al (2008) Use of light-emitting diode fluorescence microscopy to detect acid-fast bacilli in sputum. Clin Infect Dis 47(2):203–207CrossRefGoogle Scholar
  15. 15.
    Miller AR et al (2010) Portable, battery-operated, low-cost, bright field and fluorescence microscope. PLoS One 5(8):e11890CrossRefGoogle Scholar
  16. 16.
    Tapley A et al (2013) Mobile digital fluorescence microscopy for diagnosis of tuberculosis. J Clin Microbiol 51(6):1774–1778CrossRefGoogle Scholar
  17. 17.
    Urdániz E et al (2016) Antimicrobial Agents and Chemotherapy 60(5):3253–3256CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Departamento de Química Biológica, Facultad de Ciencias Exactas y NaturalesUniversidad de Buenos Aires, IQUIBICEN-CONICETBuenos AiresArgentina
  2. 2.Laboratorio “Bacteriófagos y Aplicaciones Biotecnológicas”, Departamento de Química Biológica, FCEyN, UBACiudad UniversitariaCiudad Autónoma de Buenos AiresArgentina
  3. 3.Department of Biological Sciences, Pittsburgh Bacteriophage InstituteUniversity of PittsburghPittsburghUSA

Personalised recommendations