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2017, Número 5

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Gac Med Mex 2017; 153 (5)


Detección molecular de especies de Candida en especímenes de pacientes hospitalizados

Camacho-Cardoso JL, Martínez-Rivera MÁ, Manzano-Gayosso P, Méndez-Tovar LJ, López-Martínez R, Hernández-Hernández F

Idioma: Español
Referencias bibliográficas: 64
Paginas: 581-589
Archivo PDF: 313.14 Kb.


RESUMEN

Objetivo: Detectar las especies de Candida más frecuentes en especímenes de pacientes hospitalizados en diferentes centros médicos de la Ciudad de México, con sospecha de infección fúngica. Métodos: Los especímenes fueron cultivados en agar dextrosa Sabouraud a 28 °C por 72 horas. Además, se realizó extracción de ADN. Los aislados fueron cultivados en CHROMagar Candida™ a 37 °C por 72 horas. La identificación molecular se efectuó mediante reacción en cadena de la polimerasa (PCR), utilizando iniciadores específicos para cuatro especies. Resultados: Se procesaron 81 especímenes que incluyeron líquidos de lavado bronquial, pleural, cefalorraquídeo, peritoneal, ascitis y biliar; sangre, esputo, médula ósea, cánula orotraqueal y ganglio. Por cultivo, 30 (37%) muestras fueron positivas, y por PCR, 41 (50.6%). Por PCR, la frecuencia de especies detectadas fue C. albicans 82.9%, C. tropicalis 31.7%, C. glabrata 24.4% y C. parapsilosis 4.9%. En el 34.1% de los especímenes se detectó mezcla de especies, indicando coinfección: dos especies en cinco especímenes (C. albicans-C. tropicalis y C. albicans- C. glabrata) y tres especies en tres especímenes (C. albicans-C. glabrata-C. tropicalis). Conclusiones: La PCR es una herramienta útil para detectar las especies de Candida comúnmente causantes de infección en pacientes hospitalizados, no requiere cultivo si se parte directamente del espécimen clínico y favorece el diagnóstico temprano de candidiasis invasiva.


REFERENCIAS (EN ESTE ARTÍCULO)

  1. Linton CJ, Borman AM, Cheung G, et al. Molecular identification of unusual pathogenic yeast isolates by large ribosomal subunit gene sequencing: 2 years of experience at the United Kingdom mycology reference laboratory. J Clin Microbiol. 2007;45:1152-8.

  2. Falagas ME, Roussos N, Vardakas KZ. Relative frequency of albicans and the various non-albicans Candida spp among candidemia isolates from inpatients in various parts of the world: a systematic review. Int J Infect Dis. 2010;14: e954-66.

  3. Aittakorpi A, Kuusela P, Koukila-Kahkola P, et al. Accurate and rapid identification of Candida spp. frequently associated with fungemia by using PCR and the microarray-based prove-it sepsis assay. J Clin Microbiol. 2012;50:3635-40

  4. Sardi JSO, Scorzoni L, Bernardi T, et al. Candida species: current epidemiology, pathogenicity, biofilm formation, natural antifungal products and new therapeutic options. J Med Microbiol. 2013;62:10-24.

  5. Armstrong-James D. Invasive Candida species infection: the importance of adequate empirical antifungal therapy. J Antimicrob Chemother. 2007;60:459-60.

  6. Pfaller MA, Diekema DJ. Epidemiology of invasive candidiasis: a persistent public health problem. Clin Microbiol Rev. 2007;20:133-63.

  7. Guinea J. Global trends in the distribution of Candida species causing candidemia. Clin Microbiol Infect. 2014;20:5-10.

  8. Yapar N. Epidemiology and risk factors for invasive candidiasis. Ther Clin Risk Manag. 2014;10:95-105.

  9. Estrada-Barraza D, Dávalos-Martínez A, Flores-Padilla L, et al. Comparación entre métodos convencionales, CHROMagar Candida y el método de la PCR para la identificación de especies de Candida en aislamientos clínicos. Rev beroam Micol. 2011;28:36-42.

  10. Durán-Valle MT, Sanz-Rodríguez N, Muñoz-Paraíso C, et al. Identification of clinical yeasts by Vitek MS system compared with API ID 32 C. Med Mycol. 2014;52:342-9.

  11. Pontón J. Utilidad de los marcadores biológicos en el diagnóstico de la candidiasis invasora. Rev Iberoam Micol. 2009;26 8-14.

  12. Ellepola ANB, Morrison CJ. Laboratory diagnosis of invasive candidiasis. J Microbiol. 2005;43:65-84.

  13. Pincus DH, Orenga S, Chatellier S. Yeast identification − past, present, and future methods. Med Mycol. 2007;45:97-121.

  14. Lopes-Colombo A, Cortes JA, Zurita J, et al. Recommendations for the diagnosis of candidemia in Latin America. Rev Iberoam Micol. 2013; 30:150-7.

  15. McCarty TP, Pappas PG. Invasive candidiasis. Infect Dis Clin North Am. 2016;30:103-24.

  16. Luo G, Mitchell TG. Rapid identification of pathogenic fungi directly from cultures by using multiplex PCR. J Clin Microbiol. 2002;40:2860-5.

  17. Buitrago MJ, Aguado JM, Ballen A, et al. Efficacy of DNA amplification in tissue biopsy samples to improve the detection of invasive fungal disease. Clin Microbiol Infect. 2012;19:E271-77.

  18. Arvanitis M, Anagnostou T, Fuchs BB, et al. Molecular and nonmolecular diagnostic methods for invasive fungal infections. Clin Microbiol Rev. 2014;27:490-526.

  19. Blin N, Stafford DW. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976;3:2303-8.

  20. Loffler J, Hebart H, Schumacher U, et al. Comparison of different methods for extraction of DNA of fungal pathogens from cultures and blood. J Clin Microbiol. 1997;35:3311-2.

  21. Schabereiter-Gurtner C, Selitsch B, Rotter ML, et al. Development of novel real-time PCR assays for detection and differentiation of eleven medically important Aspergillus and Candida species in clinical specimens. J Clin Microbiol. 2007;45:906-14.

  22. Guery BP, Arendrup MC, Auzinger G, et al. Management of invasive candidiasis and candidemia in adult non-neutropenic intensive care unit patients: part I. Epidemiology and diagnosis. Intensive Care Med. 2009;35:55-62.

  23. Maaroufi Y, De Bruyne JM, Duchateau V, et al. Early detection and identification of commonly encountered Candida species from simulated blood cultures by using a real-time PCR-based assay. J Mol Diagnostics. 2004;6:108-14.

  24. Filioti J, Spiroglou K, Roilides E. Invasive candidiasis in pediatric intensive care patients: epidemiology, risk factors, management, and outcome. Intensive Care Med. 2007;33:1272-83.

  25. Méndez-Tovar LJ, Manzano-Gayosso P, Cumplido-Uribe C, et al. Micosis invasivas en pacientes inmunodeprimidos con fiebre de origen desconocido. Rev Med Inst Mex Seguro Soc. 2012;50:609-14.

  26. Hernández-Hernández F, Córdova-Martínez E, Manzano-Gayosso P, et al. Frecuencia de micosis en pacientes inmunosuprimidos de un hospital regional de la Ciudad de México. Salud Publica Mex. 2003;45:455-60.

  27. González GM, Elizondo M, Ayala J. Trends in species distribution and susceptibility of bloodstream isolates of Candida collected in Monterrey, Mexico, to seven antifungal agents: results of a 3-year (2004 to 2007) surveillance study. J Clin Microbiol. 2008;46:2902-5.

  28. Philip A, Odabasi Z, Matiuzzi G, et al. Syscan3, a kit for detection of anti-Candida antibodies for diagnosis of invasive candidiasis. J Clin Microbiol. 2005;43:4834-5.

  29. Lockhart SR, Iqbal N, Cleveland AA, et al. Species identification and antifungal susceptibility testing of Candida bloodstream isolates from population-based surveillance studies in two U. S. cities from 2008 to 2011. J Clin Microbiol. 2012;50:3435-42.

  30. Quindós G. Epidemiology of candidaemia and invasive candidiasis. A changing face. Rev Iberoam Micol. 2014;31:42-8.

  31. Manzano-Gayosso P, Méndez-Tovar LJ, Arenas R, et al. Levaduras causantes de onicomicosis en cuatro centros dermatológicos mexicanos y su sensibilidad antifúngica a compuestos azólicos. Rev Iberoam Micol. 2011;28:32-5.

  32. Pfaller MA, Moet GJ, Messer SA, et al. Candida bloodstream infections: comparison of species distributions and antifungal resistance patterns in community-onset and nosocomial isolates in the SENTRY antimicrobial surveillance program, 2008-2009. Antimicrob Agents Chemother. 2011;55 561-6.

  33. Linton CJ, Borman AM, Cheung G, et al. Molecular identification of unusual pathogenic yeast isolates by large ribosomal subunit gene sequencing: 2 years of experience at the United Kingdom mycology reference laboratory. J Clin Microbiol. 2007;45:1152-8.

  34. Falagas ME, Roussos N, Vardakas KZ. Relative frequency of albicans and the various non-albicans Candida spp among candidemia isolates from inpatients in various parts of the world: a systematic review. Int J Infect Dis. 2010;14: e954-66.

  35. Aittakorpi A, Kuusela P, Koukila-Kahkola P, et al. Accurate and rapid identification of Candida spp. frequently associated with fungemia by using PCR and the microarray-based prove-it sepsis assay. J Clin Microbiol. 2012;50:3635-40

  36. Sardi JSO, Scorzoni L, Bernardi T, et al. Candida species: current epidemiology, pathogenicity, biofilm formation, natural antifungal products and new therapeutic options. J Med Microbiol. 2013;62:10-24.

  37. Armstrong-James D. Invasive Candida species infection: the importance of adequate empirical antifungal therapy. J Antimicrob Chemother. 2007;60:459-60.

  38. Pfaller MA, Diekema DJ. Epidemiology of invasive candidiasis: a persistent public health problem. Clin Microbiol Rev. 2007;20:133-63.

  39. Guinea J. Global trends in the distribution of Candida species causing candidemia. Clin Microbiol Infect. 2014;20:5-10.

  40. Yapar N. Epidemiology and risk factors for invasive candidiasis. Ther Clin Risk Manag. 2014;10:95-105.

  41. Estrada-Barraza D, Dávalos-Martínez A, Flores-Padilla L, et al. Comparación entre métodos convencionales, CHROMagar Candida y el método de la PCR para la identificación de especies de Candida en aislamientos clínicos. Rev beroam Micol. 2011;28:36-42.

  42. Durán-Valle MT, Sanz-Rodríguez N, Muñoz-Paraíso C, et al. Identification of clinical yeasts by Vitek MS system compared with API ID 32 C. Med Mycol. 2014;52:342-9.

  43. Pontón J. Utilidad de los marcadores biológicos en el diagnóstico de la candidiasis invasora. Rev Iberoam Micol. 2009;26 8-14.

  44. Ellepola ANB, Morrison CJ. Laboratory diagnosis of invasive candidiasis. J Microbiol. 2005;43:65-84.

  45. Pincus DH, Orenga S, Chatellier S. Yeast identification − past, present, and future methods. Med Mycol. 2007;45:97-121.

  46. Lopes-Colombo A, Cortes JA, Zurita J, et al. Recommendations for the diagnosis of candidemia in Latin America. Rev Iberoam Micol. 2013; 30:150-7.

  47. McCarty TP, Pappas PG. Invasive candidiasis. Infect Dis Clin North Am. 2016;30:103-24.

  48. Luo G, Mitchell TG. Rapid identification of pathogenic fungi directly from cultures by using multiplex PCR. J Clin Microbiol. 2002;40:2860-5.

  49. Buitrago MJ, Aguado JM, Ballen A, et al. Efficacy of DNA amplification in tissue biopsy samples to improve the detection of invasive fungal disease. Clin Microbiol Infect. 2012;19:E271-77.

  50. Arvanitis M, Anagnostou T, Fuchs BB, et al. Molecular and nonmolecular diagnostic methods for invasive fungal infections. Clin Microbiol Rev. 2014;27:490-526.

  51. Blin N, Stafford DW. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976;3:2303-8.

  52. Loffler J, Hebart H, Schumacher U, et al. Comparison of different methods for extraction of DNA of fungal pathogens from cultures and blood. J Clin Microbiol. 1997;35:3311-2.

  53. Schabereiter-Gurtner C, Selitsch B, Rotter ML, et al. Development of novel real-time PCR assays for detection and differentiation of eleven medically important Aspergillus and Candida species in clinical specimens. J Clin Microbiol. 2007;45:906-14.

  54. Guery BP, Arendrup MC, Auzinger G, et al. Management of invasive candidiasis and candidemia in adult non-neutropenic intensive care unit patients: part I. Epidemiology and diagnosis. Intensive Care Med. 2009;35:55-62.

  55. Maaroufi Y, De Bruyne JM, Duchateau V, et al. Early detection and identification of commonly encountered Candida species from simulated blood cultures by using a real-time PCR-based assay. J Mol Diagnostics. 2004;6:108-14.

  56. Filioti J, Spiroglou K, Roilides E. Invasive candidiasis in pediatric intensive care patients: epidemiology, risk factors, management, and outcome. Intensive Care Med. 2007;33:1272-83.

  57. Méndez-Tovar LJ, Manzano-Gayosso P, Cumplido-Uribe C, et al. Micosis invasivas en pacientes inmunodeprimidos con fiebre de origen desconocido. Rev Med Inst Mex Seguro Soc. 2012;50:609-14.

  58. Hernández-Hernández F, Córdova-Martínez E, Manzano-Gayosso P, et al. Frecuencia de micosis en pacientes inmunosuprimidos de un hospital regional de la Ciudad de México. Salud Publica Mex. 2003;45:455-60.

  59. González GM, Elizondo M, Ayala J. Trends in species distribution and susceptibility of bloodstream isolates of Candida collected in Monterrey, Mexico, to seven antifungal agents: results of a 3-year (2004 to 2007) surveillance study. J Clin Microbiol. 2008;46:2902-5.

  60. Philip A, Odabasi Z, Matiuzzi G, et al. Syscan3, a kit for detection of anti-Candida antibodies for diagnosis of invasive candidiasis. J Clin Microbiol. 2005;43:4834-5.

  61. Lockhart SR, Iqbal N, Cleveland AA, et al. Species identification and antifungal susceptibility testing of Candida bloodstream isolates from population-based surveillance studies in two U. S. cities from 2008 to 2011. J Clin Microbiol. 2012;50:3435-42.

  62. Quindós G. Epidemiology of candidaemia and invasive candidiasis. A changing face. Rev Iberoam Micol. 2014;31:42-8.

  63. Manzano-Gayosso P, Méndez-Tovar LJ, Arenas R, et al. Levaduras causantes de onicomicosis en cuatro centros dermatológicos mexicanos y su sensibilidad antifúngica a compuestos azólicos. Rev Iberoam Micol. 2011;28:32-5.

  64. Pfaller MA, Moet GJ, Messer SA, et al. Candida bloodstream infections: comparison of species distributions and antifungal resistance patterns in community-onset and nosocomial isolates in the SENTRY antimicrobial surveillance program, 2008-2009. Antimicrob Agents Chemother. 2011;55 561-6.




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