DIFFERENCES BETWEEN AGAR DILUTION, BROTH MACRODILUTION (CLSI M27-A3) AND E TEST (AB BIODISK) FOR FLUCONAZOLE RESISTANCE IN CLINICAL CANDIDA ALBICANS ISOLATES

Authors

  • E. Robledo-Leal Unidad de Manipulación Genética, Laboratorio de Micología y Fitopatología, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León (San Nicolás de Los Garza, N.L., México)
  • J. M. Adame-Rodriguez Unidad de Manipulación Genética, Laboratorio de Micología y Fitopatología, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León (San Nicolás de Los Garza, N.L., México)
  • G. M. González Departamento de Microbiología, Facultad de Medicina, Universidad Autónoma de Nuevo León (Monterrey, N.L., México) https://orcid.org/0000-0002-4434-2534
  • E. T. Aréchiga-Carvajal Unidad de Manipulación Genética, Laboratorio de Micología y Fitopatología, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León (San Nicolás de Los Garza, N.L., México)

Abstract

Ensayos de susceptibilidad realizadas por dilución en agar (AD) se compararon con M27-A3 macrodilución de caldo (BMD) metodología (Clinical Laboratory Standards Institute y) y la prueba E (AB BIODISK) con el fin de determinar la viabilidad y la fiabilidad de dilución en agar como una prueba de unexpensive para ensayos de susceptibilidad a fluconazol (FLC) contra Candida albicans. Un total de 40 cepas fueron utilizados. Todos los aislamientos resultaron en un acuerdo para la DMO y la prueba de evaluación. 19 aislados mostraron concordancia entre los tres métodos. De los 21 resultados divergentes, 20 resultaron en una mayor MIC para la EA y 15 de ellos fueron ≥ 64 mg / ml. La densidad de células debido a un contacto proporcional de las células con el fármaco en el medio puede ser responsable de los resultados obtenidos y la divergencia de AD en comparación con las otras pruebas. AD no son capaces de sustituir ni la DMO ni prueba E, pero puede ser útil como una prueba de resistencia a la tensión preliminar para la clasificación de susceptibilidad en la investigación de laboratorio.

 

Abstract
Susceptibility assays performed by agar dilution (AD) were compared to M27-A3 broth macrodilution (BMD) methodology (Clinical Laboratory and Standards Institute) and E test (AB BIODISK) in order to determine the feasibility and reliability of agar dilution as an unexpensive test for fluconazole (FLC) susceptibility assays against Candida albicans. A total of 40 strains were used. All of the isolates resulted in agreement for BMD and E test. 19 isolates showed agreement between all three methods. Of the 21 divergent results, 20 resulted in higher MIC for AD and 15 of those were ≥64 μg/ml. Cell density due to proportional contact of cells with the drug in medium may be responsible for the results obtained and the divergence of AD compared to the other tests. AD can’t reliably substitute neither BMD nor E test, but it may be helpful as a preliminary resistance test for strain susceptibility classification in laboratory research.

 

Palabras clave: Candida albicans, hongo, resitencia de drogas

Downloads

Download data is not yet available.

References

de Nadal E., F Calero, J Ramos, and J Arino. 1999. Biochemical and genetic analyses of the role of yeast casein kinase 2 in salt tolerance. J. Bacteriol; 181:6456-6462.

Gaxiola R, IF De Larrinoa, JM Villalba and R Serrano. 1992. A novel and conserved salt-induced protein is an important determinant of salt tolerance in yeast. EMBO Journal; 11:3157–3164.

Ruiz A, L Yenush and J Arino 2003. Regulation of ENA1 Na+-ATPase Gene Expression by the Ppz1 Protein Phosphatase is Mediated by the Calcineurin Pathway. Eukaryotic Cell; 2: 937-948.

National Committee for Clinical Laboratory Standards. 2002. Reference method for broth dilution antifungal susceptibility testing of yeasts. Approved standard, 2nd ed. M27-A2. National Committee for Clinical Laboratory Standards, Wayne, Pa.

Gaxiola R et al, Op. cit.

Seneviratne CJ, LJ Jin, YH Samaranayake and LP Samaranayake 2008. Cell Density and Cell Aging as Factors Modulating Antifungal Resistance of Candida albicans Biofilms. Antimicrob Agents Chemother; 52(9):3254-66.

Palani P, M Satish, and W LaJean 2007. Role for Cell Density in Antifungal Drug Resistance in Candida albicansBiofilms. Antimicrob Agents Chemother; 51(7):2454-63.

Enjalbert B and M Whiteway 2005. Release from Quorum-Sensing Molecules Triggers Hyphal Formation duringCandida albicans Resumption of Growth. Eukaryot Cell; 4(7): 1203–1210.

Nickerson, KW, AL Atkin, and JM Hornby. 2006. Quorum sensing in dimorphic fungi: farnesol and beyond. Appl. Environ. Microbiol. 72:3805-3813.

Jia XM, ZP Ma, Y Jia, PH Gao, JD Zhang, Y Wang, YG Xu, L Wang, YY Cao, YB Cao, LX Zhang and YY Jiang 2008. RTA2, a novel gene involved in azole resistance in Candida albicans. Biochem Biophys Res Commun; 373(4):631-6.

Dunkel N, TT Liu, KS Barker, R Homayouni, J Morschhäuser and PD Rogers 2008. A gain-of-function mutation in the transcription factor Upc2p causes upregulation of ergosterol biosynthesis genes and increased fluconazole resistance in a Eukaryot Cell;7(7):1180-90.

Ferrari S, F Ischer, D Calabrese, B Posteraro, M Sanguinetti, G Fadda, B Rohde, C Bauser, O Bader and D Sanglard 2009.Gain of function mutations in CgPDR1 of Candida glabrata not only mediate antifungal resistance but also enhance virulence. PLoS Pathog; 5(1):e1000268.

Jia XM, et al, Op.cit.

Shukla S, SV Ambudkar and R Prasad 2004. Substitution of threonine-1351 in the multidrug transporter Cdr1p of Candida albicans results in hypersusceptibility to antifungal agents and threonine-1351 is essential for synergic effects of calcineurin inhibitor FK520. J Antimicrob Chemother; 54(1):38-45.

Vandeputte P, G Tronchin, G Larcher, E Ernoult, T Bergès, D Chabasse et al 2008. A nonsense mutation in the ERG6 gene leads to reduced susceptibility to polyenes in a clinical isolate of Candida glabrata. Antimicrob Agents Chemother; 52(10):3701-9.

Downloads

Published

2012-07-10

How to Cite

Robledo-Leal, E., Adame-Rodriguez, J. M., González, G. M., & Aréchiga-Carvajal, E. T. (2012). DIFFERENCES BETWEEN AGAR DILUTION, BROTH MACRODILUTION (CLSI M27-A3) AND E TEST (AB BIODISK) FOR FLUCONAZOLE RESISTANCE IN CLINICAL CANDIDA ALBICANS ISOLATES. RESPYN Revista Salud Pública Y Nutrición, 13(2). Retrieved from https://respyn.uanl.mx/index.php/respyn/article/view/306

Issue

Vol. 13 No. 2 (2012): ABR-JUN 2012

Section

Artículo Original

License

Copyright (c) 2012 E. Robledo-Leal, J. M. Adame-Rodriguez, G. M. González, E. T. Aréchiga-Carvajal

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

The rights of the work belong to the author or authors, however, by sending it for publication in the Public Health and Nutrition Magazine of the Faculty of Public Health and Nutrition of the Autonomous University of Nuevo León, they grant the right for its first publication in between electronic, and possibly, in print to the Public Health and Nutrition Magazine. The license used is the Creative Commons attribution, which allows third parties to use what is published whenever the authorship of the work is mentioned and the first publication that is in the Public Health and Nutrition Magazine. Likewise, the author or authors will take into account that it will not be allowed to send the publication to any other journal, regardless of the format. The authors will be able to make other independent and additional contractual agreements for the non-exclusive distribution of the version of the article published in the Public Health and Nutrition Magazine (e.g., institutional repository or publication in a book) provided they clearly state that The work was published for the first time in the Public Health Magazine, Magazine of the Faculty of Public Health and Nutrition of the Autonomous University of Nuevo León.