Información del artículo
El desarrollo de pruebas moleculares de diagnóstico micológico no ha dejado de aumentar en los últimos años debido al incremento en la prevalencia de las infecciones fúngicas y a la lentitud en alcanzar un diagnóstico mediante las técnicas microbiológicas clásicas. Estas pruebas moleculares están diseñadas para resolver los siguientes aspectos del diagnóstico micológico: a) identificación de especie mediante secuenciación de dianas taxonómicamente relevantes; b) diagnóstico clínico precoz de las infecciones fúngicas; c) detección de los mecanismos de resistencia a los antifúngicos, y d) tipificación subespecífica de cepas clínicas. Actualmente, estas metodologías siguen circunscritas a centros de referencia tecnológicamente avanzados. No obstante, parece probable que, en breve, algunas de estas técnicas estarán disponibles en los laboratorios asistenciales.
Palabras clave:
Técnicas moleculares
Infecciones fúngicas
Diagnóstico
An increasing number of molecular techniques for the diagnosis of fungal infections have been developed in the last few years, due to the growing prevalence of mycoses and the length of time required for diagnosis when classical microbiological methods are used. These methods are designed to resolve the following aspects of mycological diagnosis: a) Identification of fungi to species level by means of sequencing relevant taxonomic targets; b) early clinical diagnosis of invasive fungal infections; c) detection of molecular mechanisms of resistance to antifungal agents; and d) molecular typing of fungi. Currently, these methods are restricted to highly developed laboratories. However, some of these techniques will probably be available in daily clinical practice in the near future.
Key words:
Molecular techniques
Fungal infections
Diagnosis
El Texto completo está disponible en PDF
Bibliografía
[1.]
B. Almirante, D. Rodríguez, B.J. Park, M. Cuenca-Estrella, A.M. Planes, M. Almela, et al.
Epidemiology and predictors of mortality in cases of Candida bloodstream infection: results from population-based surveillance, Barcelona, Spain, from 2002 to 2003.
J Clin Microbiol, 43 (2005), pp. 1829-1835
[2.]
S. Ascioglu, J. Rex, B. de Pauw, J. Bennett, J. Bille, F. Crokaert, et al.
Defining opportunistic invasive fungal infections in immunocompromised patients with cancer and hematopoietic stem cell transplants: an international consensus.
Clin Infect Dis, 34 (2002), pp. 7-14
[3.]
K. Garey, M. Rege, M. Pai, D. Mingo, K. Suda, R. Turpin, et al.
Time to initiation of fluconazole therapy impacts mortality in patients with candidemia: a multi-institutional study.
Clin Infect Dis, 43 (2006), pp. 25-31
[4.]
J. Gavaldà, O. Len, R. San Juan, J.m. Aguado, J. Fortun, C. Lumbreras, et al.
Risk Factors For Invasive Aspergillosis In Solid-Organ Transplant Recipients: A Case-Control Study.
Clin Infect Dis, 41 (2005), pp. 52-59
[5.]
M. Cuenca-Estrella, A. Gómez-López, E. Mellado, M. Buitrago, A. Monzón, Jl. Rodríguez-Tudela.
Head-to-head comparison of the activities of currently available antifungal agents against 3,378 Spanish clinical isolates of yeasts and filamentous fungi.
Antimicrob Agents Chemother, 50 (2006), pp. 917-921
[6.]
A. Alastruey-Izquierdo, M. Cuenca-Estrella, A. Monzón, Jl. Rodríguez-Tudela.
Prevalence and susceptibility testing of new species of Pseudallescheria and Scedosporium in a collection of clinical mold isolates.
Antimicrob Agents Chemother, 51 (2007), pp. 748-751
[7.]
A. Alastruey-Izquierdo, M. Cuenca-Estrella, A. Monzón, E. Mellado, Jl. Rodríguez-Tudela.
Antifungal susceptibility profile of clinical Fusarium spp. isolates identified by molecular methods.
J Antimicrob Chemother, 61 (2008), pp. 805-809
[8.]
A. Gómez-López, A. Alastruey-Izquierdo, D. Rodríguez, B. Almirante, A. Pahissa, Jl. Rodríguez-Tudela, et al.
Prevalence and susceptibility profile of Candida metapsilosis and Candida orthopsilosis: results from population-based surveillance of candidemia in Spain.
Antimicrob Agents Chemother, 52 (2008), pp. 1506-1509
[9.]
K. Nagao, T. Ota, A. Tanikawa, Y. Takae, T. Mori, S. Udagawa, et al.
Genetic identification and detection of human pathogenic rhizopus species, a major mucormycosis agent, by multiplex PCR based on internal transcribed spacer region of rrna gene.
J Dermatol Sci, 39 (2005), pp. 23-31
[10.]
T. Díaz-Guerra, E. Mellado, M. Cuenca Estrella, F. Laguna, Jl. Rodríguez-Tudela.
Molecular characterization by PCR-fingerprinting of Candida dubliniensis strains isolated from two HIV-positive patients in Spain.
Diagn Microbiol Infect Dis, 35 (1999), pp. 113-119
[11.]
J. Bishop, N. Chase, S. Magill, C. Kurtzman, M. Fiandaca, W. Merz.
Candida bracarensis detected among isolates of Candida glabrata by peptide nucleic acid fluorescence in situ hybridization: susceptibility data and documentation of presumed infection.
J Clin Microbiol, 46 (2008), pp. 443-446
[12.]
A.M. Borman, R. Petch, C.J. Linton, M.D. Palmer, P.D. Bridge, E.M. Johnson.
Candida nivariensis, an emerging pathogenic fungus with multidrug resistance to antifungal agents.
J Clin Microbiol, 46 (2008), pp. 933-938
[13.]
Jl. Rodríguez-Tudela, T.M. Díaz-Guerra, E. Mellado, V. Cano, C. Tapia, A. Perkins, et al.
Susceptibility patterns and molecular identification of Trichosporon species.
Antimicrob Agents Chemother, 49 (2005), pp. 4026-4034
[14.]
I. Gadea, M. Cuenca-Estrella.
Recomendaciones para el diagnóstico micológico y los estudios de sensibilidad a los antifúngicos.
Enferm Infecc Microbiol Clin, 22 (2004), pp. 32-39
[15.]
K. Hoffmann, S. Discher, K. Voigt.
Revision of the genus Absidia (mucorales, zygomycetes) based on physiological, phylogenetic, and morphological characters; thermotolerant Absidia spp. form a coherent group, Mycocladiaceae fam. nov.
Mycological Research, 111 (2007), pp. 1169-1183
[16.]
T. Ohst, S. de Hoog, W. Presber, V. Stavrakieva, Y. Graser.
Origins of microsatellite diversity in the Trichophyton rubrum-T. violaceum clade (dermatophytes).
J Clin Microbiol, 42 (2004), pp. 4444-4448
[17.]
A. Kaszubiak, S. Klein, G.S. de Hoog, Y. Grsser.
Population structure and evolutionary origins of Microsporum canis, M. ferrugineum and M. audouinii Infection.
Genetics and Evolution, 4 (2004), pp. 179-186
[18.]
L. Alcazar-Fuoli, E. Mellado, A. Alastruey-Izquierdo, M. Cuenca-Estrella, Jl. Rodríguez-Tudela.
Aspergillus section fumigati: antifungal susceptibility patterns and sequence-based identification.
Antimicrob Agents Chemother, 52 (2008), pp. 1244-1251
[19.]
S.A. Balajee, J.L. Gribskov, E. Hanley, D. Nickle, K.A. Marr.
Aspergillus lentulus sp. nov., a new sibling species of A. fumigatus.
Eukaryot Cell, 4 (2005), pp. 625-632
[20.]
F. Gilgado, J. Cano, J. Gene, J. Guarro.
Molecular phylogeny of the Pseudallescheria boydii species complex: proposal of two new species.
J Clin Microbiol, 43 (2005), pp. 4930-4942
[21.]
F. Gilgado, J. Cano, J. Gene, D.A. Sutton, J. Guarro.
Molecular and phenotypic data supporting distinct species statuses for Scedosporium apiospermum and Pseudallescheria boydii and the proposed new species Scedosporium dehoogii.
J Clin Microbiol, 46 (2008), pp. 766-771
[22.]
A. Loy, L. Bodrossy.
Highly parallel microbial diagnostics using oligonucleotide microarrays.
Clin Chim Acta, 363 (2006), pp. 106-119
[23.]
M.J. Buitrago, A. Gómez-López, E. Mellado, Jl. Rodríguez-Tudela, M. Cuenca-Estrella.
Detection of Aspergillus spp. by real-time PCR in a murine model of pulmonary infection.
Enferm Infecc Microbiol Clin, 23 (2005), pp. 464-468
[24.]
J.P. Donnelly.
Polymerase chain reaction for diagnosing invasive aspergillosis: getting closer but still a ways to go.
Clin Infect Dis, 42 (2006), pp. 487-489
[25.]
A. Gómez-López, M.T. Martín-Gómez, P. Martín-Dávila, P. López-Onrubia, J. Gavaldà, J. Fortún, et al.
Detection of fungal DNA by real-time polymerase chain reaction: evaluation of 2 methodologies in experimental pulmonary aspergillosis.
Diagn Microbiol Infect Dis, 56 (2006), pp. 387-393
[26.]
R. Mcmullan, L. Metwally, P.V. Coyle, S. Hedderwick, B. Mccloskey, H.j. O’neill, et al.
A prospective clinical trial of a real-time polymerase chain reaction assay for the diagnosis of candidemia in nonneutropenic, critically ill adults.
Clin Infect Dis, 46 (2008), pp. 890-896
[27.]
S. Osorio, Cr. de la, M.C. Monteserin, R. Granados, F. Ona, Jl. Rodríguez-Tudela, et al.
Recurrent disseminated skin lesions due to Metarrhizium anisopliae in an adult patient with acute myelogenous leukemia.
J Clin Microbiol, 45 (2007), pp. 651-655
[28.]
H.H. Larsen, J.A. Kovacs, F. Stock, V.H. Vestereng, B. Lundgren, S.H. Fischer, et al.
Development of a rapid real-time pcr assay for quantitation of Pneumocystis carinii F. sp. carinii.
J Clin Microbiol, 40 (2002), pp. 2989-2993
[29.]
H.H. Larsen, M.L. von Linstow, B. Lundgren, B. Hogh, H. Westh, J.D. Lundgren.
Primary Pneumocystis infection in infants hospitalized with acute respiratory tract infection.
Emerg Infect Dis, 13 (2007), pp. 66-72
[30.]
M.J. Buitrago, J. Berenguer, E. Mellado, Jl. Rodríguez-Tudela, M. Cuenca-Estrella.
Detection of imported histoplasmosis in serum of HIV-infected patients using a real-time pcr-based assay.
Eur J Clin Microbiol Infect Dis, 25 (2006), pp. 665-668
[31.]
M.J. Buitrago, A. Gómez-López, A. Monzón, Jl. Rodríguez-Tudela, M. Cuenca-Estrella.
Assessment of a quantitative PCR method for clinical diagnosis of imported histoplasmosis.
Enferm Infecc Microbiol Clin, 25 (2007), pp. 16-22
[32.]
S.H. Marques da Silva, F. Queiroz-Telles, A.L. Colombo, M.H. Blotta, J.D. Lopes, D.C. Pires.
Monitoring Gp43 antigenemia in paracoccidioidomycosis patients during therapy.
J Clin Microbiol, 42 (2004), pp. 2419-2424
[33.]
G. Ricci, I.D. da Silva, A. Sano, R.C. Borra, M. Franco.
Detection of Paracoccidioides brasiliensis by PCR in biopsies from patients with paracoccidioidomycosis: correlation with the histopathological pattern.
Pathologica, 99 (2007), pp. 41-45
[34.]
S.V. Balashov, R. Gardiner, S. Park, D.S. Perlin.
Rapid, high-throughput, multiplex, real-time PCR for identification of mutations in the Cyp51a gene of Aspergillus fumigatus that confer resistance to itraconazole.
J Clin Microbiol, 43 (2005), pp. 214-222
[35.]
G. García-Effron, A. Dilger, L. Alcázar-Fuoli, S. Park, E. Mellado, D.S. Perlin.
Rapid detection of triazole antifungal resistance in Aspergillus fumigatus.
J Clin Microbiol, 46 (2008), pp. 1200-1206
[36.]
E. Mellado, G. García-Effron, L. Alcázar-Fuoli, W.J. Melchers, P.E. Verweij, M. Cuenca-Estrella, et al.
A new Aspergillus fumigatus resistance mechanism conferring in vitro cross-resistance to azole antifungals involves a combination of Cyp51a alterations.
Antimicrob Agents Chemother, 51 (2007), pp. 1897-1904
[37.]
D. Sanglard, F. Ischer, D. Calabrese, M. Micheli, J. Bille.
Multiple resistance mechanisms to azole antifungals in yeast clinical isolates.
Drug Resist Update, 1 (1998), pp. 255-265
[38.]
S.V. Balashov, S. Park, D.S. Perlin.
Assessing resistance to the echinocandin antifungal drug caspofungin in Candida albicans by profiling mutations in Fks1.
Antimicrob Agents Chemother, 50 (2006), pp. 2058-2063
[39.]
M. Cuenca-Estrella, E. Mellado.
Are molecular techniques useful in aspergillosis surveillance and control.
Enferm Infecc Microbiol Clin, 21 (2003), pp. 469-471
[40.]
M.C. Galhardo, R.M. de Oliveira, A.C. Valle, R.A. Paes, P.M. Silvatavares, A. Monzón, et al.
Molecular epidemiology and antifungal susceptibility patterns of sporothrix schenckii isolates from a cat-transmitted epidemic of sporotrichosis in Rio de Janeiro, Brazil.
Med Mycol, 46 (2008), pp. 141-151
[41.]
F.C. Odds, M.E. Bougnoux, D.J. Shaw, J.M. Bain, A.D. Davidson, D. Diogo, et al.
Molecular phylogenetics of Candida albicans.
Eukaryot Cell, 6 (2007), pp. 1041-1052
[42.]
T.M. Díaz-Guerra, E. Mellado, M. Cuenca-Estrella, L. Gaztelurrutia, J.I. Navarro, J.L. Tudela.
Genetic similarity among one Aspergillus flavus strain isolated from a patient who underwent heart surgery and two environmental strains obtained from the operating room.
J Clin Microbiol, 38 (2000), pp. 2419-2422
[43.]
I. Gadea, M. Cuenca-Estrella, E. Prieto, T.M. Díaz-Guerra, J.I. García-Cia, E. Mellado, et al.
Genotyping and antifungal susceptibility profile of Dipodascus capitatus isolates causing disseminated infection in seven hematological patients of a tertiary hospital.
J Clin Microbiol, 42 (2004), pp. 1832-1836
Copyright © 2008. Elsevier España S.L.. Todos los derechos reservados
