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Published in: European Journal of Clinical Microbiology & Infectious Diseases 10/2015

01-10-2015 | Article

Rapid susceptibility testing for slowly growing nontuberculous mycobacteria using a colorimetric microbial viability assay based on the reduction of water-soluble tetrazolium WST-1

Authors: T. Tsukatani, H. Suenaga, M. Shiga, T. Ikegami, M. Ishiyama, T. Ezoe, K. Matsumoto

Published in: European Journal of Clinical Microbiology & Infectious Diseases | Issue 10/2015

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Abstract

Rapid susceptibility testing for slowly growing nontuberculous mycobacteria (NTM) using a colorimetric microbial viability assay based on the reduction of the water-soluble tetrazolium salt {2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-1)} using 2,3,5,6-tetramethyl-1,4-benzoquinone as an electron mediator was developed. Using the Clinical and Laboratory Standards Institute (CLSI) method, a long-term incubation time (7–14 days) was required to determine the minimum inhibitory concentrations (MICs) of the slowly growing NTM. The MICs for a variety of different antibiotics against the slowly growing NTM were determined by the WST-1 colorimetric method and compared with those obtained using the broth microdilution methods approved by the CLSI. Good agreement was found between the MICs determined after 3–4 days using the WST-1 colorimetric method and those obtained after 10–14 days using the broth microdilution method. The results suggest that the WST-1 colorimetric assay is a useful method for the rapid determination of the MICs for the slowly growing NTM.
Literature
1.
go back to reference Covert TC, Rodgers MR, Reyes AL, Stelma GN Jr (1999) Occurrence of nontuberculous mycobacteria in environmental samples. Appl Environ Microbiol 65:2492–2496PubMedCentralPubMed Covert TC, Rodgers MR, Reyes AL, Stelma GN Jr (1999) Occurrence of nontuberculous mycobacteria in environmental samples. Appl Environ Microbiol 65:2492–2496PubMedCentralPubMed
4.
go back to reference Fernandez-Rendon E, Cerna-Cortes JF, Ramirez-Medina MA, Helguera-Repetto AC, Rivera-Gutierrez S, Estrada-Garcia T, Gonzalez-y-Merchand JA (2012) Mycobacterium mucogenicum and other non-tuberculous mycobacteria in potable water of a trauma hospital: a potential source for human infection. J Hosp Infect 80:74–76. doi:10.1016/j.jhin.2011.10.003 CrossRefPubMed Fernandez-Rendon E, Cerna-Cortes JF, Ramirez-Medina MA, Helguera-Repetto AC, Rivera-Gutierrez S, Estrada-Garcia T, Gonzalez-y-Merchand JA (2012) Mycobacterium mucogenicum and other non-tuberculous mycobacteria in potable water of a trauma hospital: a potential source for human infection. J Hosp Infect 80:74–76. doi:10.​1016/​j.​jhin.​2011.​10.​003 CrossRefPubMed
5.
go back to reference Shitrit D, Baum GL, Priess R, Lavy A, Shitrit ABG, Raz M, Shlomi D, Daniele B, Kramer MR (2006) Pulmonary Mycobacterium kansasii infection in Israel, 1999–2004: clinical features, drug susceptibility, and outcome. Chest 129:771–776. doi:10.1378/chest.129.3.771 CrossRefPubMed Shitrit D, Baum GL, Priess R, Lavy A, Shitrit ABG, Raz M, Shlomi D, Daniele B, Kramer MR (2006) Pulmonary Mycobacterium kansasii infection in Israel, 1999–2004: clinical features, drug susceptibility, and outcome. Chest 129:771–776. doi:10.​1378/​chest.​129.​3.​771 CrossRefPubMed
6.
go back to reference Winthrop KL, McNelley E, Kendall B, Marshall-Olson A, Morris C, Cassidy M, Saulson A, Hedberg K (2010) Pulmonary nontuberculous mycobacterial disease prevalence and clinical features: an emerging public health disease. Am J Respir Crit Care Med 182:977–982. doi:10.1164/rccm.201003-0503OC CrossRefPubMed Winthrop KL, McNelley E, Kendall B, Marshall-Olson A, Morris C, Cassidy M, Saulson A, Hedberg K (2010) Pulmonary nontuberculous mycobacterial disease prevalence and clinical features: an emerging public health disease. Am J Respir Crit Care Med 182:977–982. doi:10.​1164/​rccm.​201003-0503OC CrossRefPubMed
7.
go back to reference Maekawa K, Ito Y, Hirai T, Kubo T, Imai S, Tatsumi S, Fujita K, Takakura S, Niimi A, Iinuma Y, Ichiyama S, Togashi K, Mishima M (2011) Environmental risk factors for pulmonary Mycobacterium avium-intracellulare complex disease. Chest 140:723–729. doi:10.1378/chest.10-2315 CrossRefPubMed Maekawa K, Ito Y, Hirai T, Kubo T, Imai S, Tatsumi S, Fujita K, Takakura S, Niimi A, Iinuma Y, Ichiyama S, Togashi K, Mishima M (2011) Environmental risk factors for pulmonary Mycobacterium avium-intracellulare complex disease. Chest 140:723–729. doi:10.​1378/​chest.​10-2315 CrossRefPubMed
8.
go back to reference Huang CT, Tsai YJ, Wu HD, Wang JY, Yu CJ, Lee LN, Yang PC (2012) Impact of non-tuberculous mycobacteria on pulmonary function decline in chronic obstructive pulmonary disease. Int J Tuberc Lung Dis 16:539–545. doi:10.5588/ijtld.11.0412 CrossRefPubMed Huang CT, Tsai YJ, Wu HD, Wang JY, Yu CJ, Lee LN, Yang PC (2012) Impact of non-tuberculous mycobacteria on pulmonary function decline in chronic obstructive pulmonary disease. Int J Tuberc Lung Dis 16:539–545. doi:10.​5588/​ijtld.​11.​0412 CrossRefPubMed
9.
go back to reference Horsburgh CR Jr, Gettings J, Alexander LN, Lennox JL (2001) Disseminated Mycobacterium avium complex disease among patients infected with human immunodeficiency virus, 1985–2000. Clin Infect Dis 33:1938–1943. doi:10.1086/324508 CrossRefPubMed Horsburgh CR Jr, Gettings J, Alexander LN, Lennox JL (2001) Disseminated Mycobacterium avium complex disease among patients infected with human immunodeficiency virus, 1985–2000. Clin Infect Dis 33:1938–1943. doi:10.​1086/​324508 CrossRefPubMed
12.
go back to reference Lai CC, Lee LN, Ding LW, Yu CJ, Hsueh PR, Yang PC (2006) Emergence of disseminated infections due to nontuberculous mycobacteria in non-HIV-infected patients, including immunocompetent and immunocompromised patients in a university hospital in Taiwan. J Infect 53:77–84. doi:10.1016/j.jinf.2005.10.009 CrossRefPubMed Lai CC, Lee LN, Ding LW, Yu CJ, Hsueh PR, Yang PC (2006) Emergence of disseminated infections due to nontuberculous mycobacteria in non-HIV-infected patients, including immunocompetent and immunocompromised patients in a university hospital in Taiwan. J Infect 53:77–84. doi:10.​1016/​j.​jinf.​2005.​10.​009 CrossRefPubMed
15.
go back to reference Park YS, Lee CH, Lee SM, Yang SC, Yoo CG, Kim YW, Han SK, Shim YS, Yim JJ (2010) Rapid increase of non-tuberculous mycobacterial lung diseases at a tertiary referral hospital in South Korea. Int J Tuberc Lung Dis 14:1069–1071PubMed Park YS, Lee CH, Lee SM, Yang SC, Yoo CG, Kim YW, Han SK, Shim YS, Yim JJ (2010) Rapid increase of non-tuberculous mycobacterial lung diseases at a tertiary referral hospital in South Korea. Int J Tuberc Lung Dis 14:1069–1071PubMed
16.
go back to reference Clinical and Laboratory Standards Institute (CLSI) (2011) Susceptibility testing of Mycobacteria, Nocardiae, and other aerobic Actinomycetes; Approved standard—Second edition. CLSI document M24-A2. CLSI, Wayne, PA, USA Clinical and Laboratory Standards Institute (CLSI) (2011) Susceptibility testing of Mycobacteria, Nocardiae, and other aerobic Actinomycetes; Approved standard—Second edition. CLSI document M24-A2. CLSI, Wayne, PA, USA
20.
go back to reference Moriarty F, Elborn S, Tunney M (2005) Development of a rapid colorimetric time–kill assay for determining the in vitro activity of ceftazidime and tobramycin in combination against Pseudomonas aeruginosa. J Microbiol Methods 61:171–179. doi:10.1016/j.mimet.2004.11.010 CrossRefPubMed Moriarty F, Elborn S, Tunney M (2005) Development of a rapid colorimetric time–kill assay for determining the in vitro activity of ceftazidime and tobramycin in combination against Pseudomonas aeruginosa. J Microbiol Methods 61:171–179. doi:10.​1016/​j.​mimet.​2004.​11.​010 CrossRefPubMed
21.
go back to reference De Logu A, Uda P, Pellerano ML, Pusceddu MC, Saddi B, Schivo ML (2001) Comparison of two rapid colorimetric methods for determining resistance of Mycobacterium tuberculosis to rifampin, isoniazid, and streptomycin in liquid medium. Eur J Clin Microbiol Infect Dis 20:33–39. doi:10.1007/PL00011234 CrossRefPubMed De Logu A, Uda P, Pellerano ML, Pusceddu MC, Saddi B, Schivo ML (2001) Comparison of two rapid colorimetric methods for determining resistance of Mycobacterium tuberculosis to rifampin, isoniazid, and streptomycin in liquid medium. Eur J Clin Microbiol Infect Dis 20:33–39. doi:10.​1007/​PL00011234 CrossRefPubMed
22.
go back to reference De Logu A, Pellerano ML, Sanna A, Pusceddu MC, Uda P, Saddi B (2003) Comparison of the susceptibility testing of clinical isolates of Mycobacterium tuberculosis by the XTT colorimetric method and the NCCLS standards method. Int J Antimicrob Agents 21:244–250. doi:10.1016/S0924-8579(02)00350-3 CrossRefPubMed De Logu A, Pellerano ML, Sanna A, Pusceddu MC, Uda P, Saddi B (2003) Comparison of the susceptibility testing of clinical isolates of Mycobacterium tuberculosis by the XTT colorimetric method and the NCCLS standards method. Int J Antimicrob Agents 21:244–250. doi:10.​1016/​S0924-8579(02)00350-3 CrossRefPubMed
23.
go back to reference De Logu A, Borgna R, Uda P, Sanna A, Pellerano ML, Saddi B (2003) The 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay as rapid colorimetric method for determination of antibiotic susceptibility of clinical Mycobacterium tuberculosis isolates in liquid medium. Clin Lab 49:357–365PubMed De Logu A, Borgna R, Uda P, Sanna A, Pellerano ML, Saddi B (2003) The 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay as rapid colorimetric method for determination of antibiotic susceptibility of clinical Mycobacterium tuberculosis isolates in liquid medium. Clin Lab 49:357–365PubMed
25.
26.
go back to reference Tsukatani T, Higuchi T, Suenaga H, Akao T, Ishiyama M, Ezoe T, Matsumoto K (2009) Colorimetric microbial viability assay based on reduction of water-soluble tetrazolium salts for antimicrobial susceptibility testing and screening of antimicrobial substances. Anal Biochem 393:117–125. doi:10.1016/j.ab.2009.06.026 CrossRefPubMed Tsukatani T, Higuchi T, Suenaga H, Akao T, Ishiyama M, Ezoe T, Matsumoto K (2009) Colorimetric microbial viability assay based on reduction of water-soluble tetrazolium salts for antimicrobial susceptibility testing and screening of antimicrobial substances. Anal Biochem 393:117–125. doi:10.​1016/​j.​ab.​2009.​06.​026 CrossRefPubMed
27.
go back to reference Ishiyama M, Shiga M, Sasamoto K, Mizoguchi M, He P (1993) A new sulfonated tetrazolium salt that produces a highly water-soluble formazan dye. Chem Pharm Bull 41:1118–1122CrossRef Ishiyama M, Shiga M, Sasamoto K, Mizoguchi M, He P (1993) A new sulfonated tetrazolium salt that produces a highly water-soluble formazan dye. Chem Pharm Bull 41:1118–1122CrossRef
28.
go back to reference Ishiyama M, Tominaga H, Shiga M, Sasamoto K, Ohkura Y, Ueno K (1996) A combined assay of cell viability and in vitro cytotoxicity with a highly water-soluble tetrazolium salt, neutral red and crystal violet. Biol Pharm Bull 19:1518–1520CrossRefPubMed Ishiyama M, Tominaga H, Shiga M, Sasamoto K, Ohkura Y, Ueno K (1996) A combined assay of cell viability and in vitro cytotoxicity with a highly water-soluble tetrazolium salt, neutral red and crystal violet. Biol Pharm Bull 19:1518–1520CrossRefPubMed
30.
go back to reference Mäkinen J, Marjamäki M, Haanperä-Heikkinen M, Marttila H, Endourova LB, Presnova SE, Mathys V, Bifani P, Ruohonen R, Viljanen MK, Soini H (2011) Extremely high prevalence of multidrug resistant tuberculosis in Murmansk, Russia: a population-based study. Eur J Clin Microbiol Infect Dis 30:1119–1126. doi:10.1007/s10096-011-1200-7 CrossRefPubMed Mäkinen J, Marjamäki M, Haanperä-Heikkinen M, Marttila H, Endourova LB, Presnova SE, Mathys V, Bifani P, Ruohonen R, Viljanen MK, Soini H (2011) Extremely high prevalence of multidrug resistant tuberculosis in Murmansk, Russia: a population-based study. Eur J Clin Microbiol Infect Dis 30:1119–1126. doi:10.​1007/​s10096-011-1200-7 CrossRefPubMed
31.
go back to reference Agrawal D, Udwadia ZF, Rodriguez C, Mehta A (2009) Increasing incidence of fluoroquinolone-resistant Mycobacterium tuberculosis in Mumbai, India. Int J Tuberc Lung Dis 13:79–83PubMed Agrawal D, Udwadia ZF, Rodriguez C, Mehta A (2009) Increasing incidence of fluoroquinolone-resistant Mycobacterium tuberculosis in Mumbai, India. Int J Tuberc Lung Dis 13:79–83PubMed
Metadata
Title
Rapid susceptibility testing for slowly growing nontuberculous mycobacteria using a colorimetric microbial viability assay based on the reduction of water-soluble tetrazolium WST-1
Authors
T. Tsukatani
H. Suenaga
M. Shiga
T. Ikegami
M. Ishiyama
T. Ezoe
K. Matsumoto
Publication date
01-10-2015
Publisher
Springer Berlin Heidelberg
Published in
European Journal of Clinical Microbiology & Infectious Diseases / Issue 10/2015
Print ISSN: 0934-9723
Electronic ISSN: 1435-4373
DOI
https://doi.org/10.1007/s10096-015-2438-2

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