Applications of Flow Cytometry to Clinical Microbiology

Alvarez-Barrientos, Alberto; Arroyo, Javier; Cantón, Rafael; Nombela, César; Sanchez-Perez, M.

doi:10.1128/cmr.13.2.167-195.2000

Cited by 161 publications

(126 citation statements)

References 203 publications

(299 reference statements)

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“…units (21). This equipment has been used for many purposes in microbiology, including basic research (1), to study the effects of chemical compounds on microbial growth (17,19), for quality control (21), and in pollutant degradation tests (34).…”

Section: Introductionmentioning

confidence: 99%

Comparative growth of trichoderma strains in different nutritional sources, using bioscreen c automated system

Rossi-Rodrigues¹,

Brochetto-Braga²,

Tauk-Tornisielo³

et al. 2009

Braz. J. Microbiol.

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Trichoderma is one of the fungi genera that produce important metabolites for industry. The growth of these organisms is a consequence of the nutritional sources used as also of the physical conditions employed to cultivate them. In this work, the automated Bioscreen C system was used to evaluate the influence of different nutritional sources on the growth of Trichoderma strains (T. hamatum, T. harzianum, T. viride, and T. longibrachiatum) isolated from the soil in the Juréia-Itatins Ecological Station (JIES), São Paulo StateBrazil. The cultures were grown in liquid culture media containing different carbon-(2%; w/v) and nitrogen (1%; w/v) sources at 28ºC, pH 6.5, and agitated at 150 rpm for 72 h. The results showed, as expected, that glucose is superior to sucrose as a growth-stimulating carbon source in the Trichoderma strains studied, while yeast extract and tryptone were good growth-stimulating nitrogen sources in the cultivation of T. hamatum and T. harzianum.

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Section: Introductionmentioning

confidence: 99%

Comparative growth of trichoderma strains in different nutritional sources, using bioscreen c automated system

Rossi-Rodrigues¹,

Brochetto-Braga²,

Tauk-Tornisielo³

et al. 2009

Braz. J. Microbiol.

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“…Furthermore, flow cytometry, a powerful tool in bacteriology [4], was used to study the antibacterial effect of lactacin F and nisin separately and in combination.…”

Section: Introductionmentioning

confidence: 99%

Bacterial membrane injuries induced by lactacin F and nisin

et al. 2002

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The combined action of nisin and lactacin F, two bacteriocins produced by lactic acid bacteria, is additive. In this report, the basis of this effect is examined. Channels formed by lactacin F were studied by experiments using planar lipid bilayers, and bactericidal effects were analyzed by flow cytometry. Lactacin F produced pores with a conductance of 1 ns in black lipid bilayers in 1 mM KCl at 10 mV at 20°C. Pore formation was strongly dependent on voltage. Although lactacin F formed pores at very low potential (10 mV), the dependence was exponential above 40 mV. The injuries induced by nisin and lactacin F in the membranes of Lactobacillus helveticus produced different flow cytometric profiles. Probably, when both bacteriocins are present, each acts separately; their cooperation may be due to an increase in the number of single membrane injuries.

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“…Once we can assure biosafety by adding formaldehyde before acquiring results in the flow cytometer, further containment would not be required. Microbiological use of a flow cytometer should be considered (even if only for screening of drug resistance to help avoiding the spread of drug-resistant tuberculosis) (1,4,5,16). We could confirm the feasibility of use of flow cytometry with clinical samples of M. tuberculosis in 24 h.…”

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confidence: 74%

Comparison of Flow Cytometric and Alamar Blue Tests with the Proportional Method for Testing Susceptibility of Mycobacterium tuberculosis to Rifampin and Isoniazid

et al. 2004

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The performance of flow cytometry and the microplate Alamar Blue assay in determining susceptibility of Mycobacterium tuberculosis was assessed by testing 150 Brazilian isolates. The overall agreement was 97.3 and 98% for isoniazid and 94.7 and 100% for rifampin by flow cytometry and MABA, respectively. This study was entirely done in a developing country.Many developing countries have serious difficulties obtaining drug susceptibility information for Mycobacterium tuberculosis isolates for financial or technical reasons. Treatment of tuberculosis without susceptibility information increases the risk of treatment failure and of the spread of resistant strains as well as the risk of the development of resistance to additional drugs (2,8).The commonly used agar proportion method for mycobacterial susceptibility testing requires a 3-to 4-week period of incubation before a pattern of susceptibility is established (3, 9). Flow cytometry that relies on fluoresceine diacetate (FDA) for detection has been used to perform susceptibility testing of M. tuberculosis and can yield results within 24 h. The inhibition by rifampin (RIF) and isoniazid (INH) of the ability of viable M. tuberculosis to hydrolyze FDA can be measured by flow cytometry. In addition, multiplication of the mycobacteria is not required (10,11,15).The microplate Alamar Blue assay (MABA) (7) has been reported to show very good correlations with the proportional and BACTEC methods. MABA is a resarzurin-based oxidation-reduction indicator which measures colorimetric drug MICs for M. tuberculosis for up to 7 days. In this study, we compared the flow cytometric and MABA tests with the standard proportional method to assess INH and RIF susceptibilities of 150 clinical isolates from a community in Rio de Janeiro, Brazil; 100 isolates were susceptible to both drugs, 50 were resistant to INH, and 37 of these 50 were also RIF resistant (multidrug resistant).The proportional method was performed according to the method of Canetti, Rist, and Grosset (3). The results obtained by the proportional method were used as a reference to compare the results of flow cytometry and MABA.For mycobacterium preparation, each isolate was grown in two Löwenstein-Jensen tubes (Difco, Detroit, Mich.) at 37°C in aerobic conditions for 30 days. After incubation, colonies were suspended in Middlebrook 7H9 medium (Difco) directly from solid medium and adjusted to a no. 1 McFarland standard (ϳ3 ϫ 10 7 CFU/ml).Flow cytometric susceptibility testing was performed according to the method of Norden et al. (11). The only modification was implemented to assure bacterial inactivation before analysis with an XL-MCL flow cytometer (Coulter, Miami, Fla.). In a previous study (data not published), we showed that M. tuberculosis cells can be killed by formaldehyde at a final concentration of 10% for 1 h.Final drug concentrations were 0.5, 1.0, and 2.0 g/ml for RIF and 0.1, 0.2, and 0.3 g/ml for INH. For each isolate the relative fluorescence value of each drug-containing sample was divided by the rela...

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Applications of Flow Cytometry to Clinical Microbiology

Cited by 161 publications

References 203 publications

Comparative growth of trichoderma strains in different nutritional sources, using bioscreen c automated system

Comparative growth of trichoderma strains in different nutritional sources, using bioscreen c automated system

Bacterial membrane injuries induced by lactacin F and nisin

Comparison of Flow Cytometric and Alamar Blue Tests with the Proportional Method for Testing Susceptibility of Mycobacterium tuberculosis to Rifampin and Isoniazid

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