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2017, Number 5

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Gac Med Mex 2017; 153 (5)

Molecular detection of candida species from hospitalized patient’s specimens

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

Language: Spanish
References: 64
Page: 581-589
PDF size: 313.14 Kb.


ABSTRACT

Objective: To identify the most frequent Candida species in specimens from patients hospitalized in different medical centers of Mexico City, with suspected fungal infection. Methods: Specimens were grown on Sabouraud dextrose agar at 28°C for 72 h. In addition, DNA was extracted. Isolates were grown on CHROMagar Candida™, at 37°C for 48 h. The molecular identification was performed by polymerase chain reaction (PCR) using primers specific for four species. Results: Eighty one specimens were processed and included: bronchial lavage, pleural, cerebrospinal, peritoneal, ascites and bile fluids; blood, sputum, bone marrow, oro-tracheal cannula and ganglion. By culture, 30 samples (37%) were positive, and by PCR, 41 (50.6%). By PCR, the frequency of species was: Candida albicans 82.9%, Candida tropicalis 31.7%, Candida glabrata 24.4%, and Candida parapsilosis 4.9%. In 34.1% of specimens a species mixture was detected suggesting a co-infection: Two species in five specimens (C. albicans-C tropicalis and C. albicans-C glabrata), and three species in three specimens (C. albicans-C. glabrata-C. tropicalis). Conclusions: The PCR is an useful tool for detection the most common Candida species causing infection in hospitalized patients, it avoids the requirement of culture weather we start from clinical specimen and it favors the early diagnosis of invasive candidiasis.


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  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.

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  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.

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  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.

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  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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