Identificación molecular de especies de candida en pacientes quemados

Medical histories of 36 burned patients with documented fungal infection,\nadmitted to the Intensive Care Unit of the Hospital Municipal de Quemados del Gobierno\nde la Ciudad de Buenos Aires between January 2011 and December 2014, were analyzed.\nA total of 52 species of Candida were identified by...

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Autor principal:	Badino Varela, María Gabriela Mercedes
Otros Autores:	Landaburu, María F.
Formato:	Tesis de maestría acceptedVersion
Lenguaje:	Español
Publicado:	Facultad de Farmacia y Bioquímica 2017
Materias:	Candida Quemaduras Infecciones fúngicas Patogénesis de quemaduras Ciencias de la vida
Acceso en línea:	http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_1901 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_1901.dir/1901.PDF
Aporte de:	Repositorio Digital de la Universidad de Buenos Aires (UBA) de Universidad de Buenos Aires

id	I28-R145-HWA_1901
record_format	dspace
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-145
collection	Repositorio Digital de la Universidad de Buenos Aires (UBA)
language	Español
orig_language_str_mv	spa
topic	Candida Quemaduras Infecciones fúngicas Patogénesis de quemaduras Ciencias de la vida
spellingShingle	Candida Quemaduras Infecciones fúngicas Patogénesis de quemaduras Ciencias de la vida Badino Varela, María Gabriela Mercedes Identificación molecular de especies de candida en pacientes quemados
topic_facet	Candida Quemaduras Infecciones fúngicas Patogénesis de quemaduras Ciencias de la vida
description	Medical histories of 36 burned patients with documented fungal infection,\nadmitted to the Intensive Care Unit of the Hospital Municipal de Quemados del Gobierno\nde la Ciudad de Buenos Aires between January 2011 and December 2014, were analyzed.\nA total of 52 species of Candida were identified by conventional and molecular methods\nfrom the culture collection stored at -20 ºC at the Laboratory of Hospital de Quemados in\nthe patients studied. The mean hospitalization period was 46.7 days, ranging from 9 to\n218 days. They belonged to females 63.9% (n = 23) and 36.1% male (n = 13). The burned\nbody surface area was estimated at 31-50% in 33.3% of the patients (p=0.03). This\npercentage was observed among the higher age group (59 years of age). Instead, patients\nin the group with a mean age of 19 years suffered burns affecting 71% - 100% of their\nbody surface area. With regard to burn depth, fourth-degree burns affected 69.4% of the\npatients, which indicates the seriousness of the injuries in the patients studied. A total of\n75% of the patients showed burns in different locations (torso, face, lower and upper\nlimbs) and only 25% showed injuries only on the lower limbs, upper limbs or torso. The\ncausative agents of burns were: combustible substances (alcohol, gasoline, explosion):\n41.7%, direct fire: 25%, heaters: 13.9%, water: 16.7% and lastly, electricity: 2.8%. It is\nremarkable that in those cases in which the burn causative agent was a combustible\nsubstance, alcohol was the prevalent agent. When studying causative agents in relation to\nage, burn injuries by fire or heaters were found mainly older patients. Conversely, burns\ncaused by alcohol, combustible substances and electricity were reported on younger\npatients. When studying causative agents in relation to sex, alcohol was found to be the\nprevalent element in females. This higher rate for women is associated with the fact that the causative agents of the burn injury are alcohol and direct fire, according to medical\nhistory including injuries by a close third party. They are the current causes of the\nfemicides reported in our country in women under 40 years of age. Electrical burns and\ngasoline burn injuries were affected males exclusively.When analyzing the correlation\nbetween the burned surface area and the cause (combustible substances vs. other agents),\nit was found that causes other than combustible substances generate smaller burn injuries\n(p= 0.01). However, second-degree burns were observed when the agent was a\ncombustible substance (alcohol, gasoline or explosion) (p=0.03), as a result of a deeper\nburn.Fifty per cent of the burned patients showed inhalation injuries (SII), with both\ncombustible substances and fire as causative agents of the burn injury. A central line was\nrequired in 88.9% of the burned patients. Femoral lines were used in 90.6% and upper\nlines (subclavian and/or jugular lines), in 9.4%. The mean number of lines used per\npatient was 5.8. Orotracheal intubation and tracheostomy were performed in 72.2% and\n41.7% of the patients, respectively. A total of 91.7 % of the patients underwent surgical\ntreatment (escharectomy: 100%, escharotomy: 42.4%, fasciotomy: 54.5% and limb\namputation: 6.1%) while 8.3% of the patients did not undergo any surgical treatment\n(only wound lavage). Following surgical treatment and, after reaching viable tissue, the\nwound was covered with skin grafts. Autograft was performed in 97.2% of the patients,\nshowing firm adherence to the underlying tissue in 75% of the cases. In twenty-five per\ncent of the unsuccessful cases, the mortality rate was 100% (p< 0.01), the main cause\nbeing the septic shock. Inotropes were required in 72.2% of the cases and anti-fungal\ndrugs in 91.7%. A total of 58.8% were initially treated with Fluconazole. The rest of the\ncases were treated with Amphotericin B Liposomal. Death occurred in 38.8% of the patients, its rate being higher among patients over 65 years of age (p= 0.018), on\ninotropes (noradrenaline) (p < 0.001), those patients suffering inhalation injury (p= 0.04),\nthose who had undergone fasciotomy (p=0.04) and those with a history or in a high-risk\ngroup (p = 0.002). Infection by bacteria occurred in 100% of the burned patients, the\nmicroorganisms being: Gram-negative bacteria such as multiresistant Acinetobacter\nbaumannii, Pseudomonas aeruginosa and carbapenemase-producing (KPC) Klebsiella\npneumoniae. Among Gram-positive bacteria, the most prevalent was methicillin-resistant\nS. Aureus (MRSA). The mean time of the onset of bacterial infections was 4 days\nfollowing admission to the Intensive Care Unit. A total of 38.9% of the patients had an\nAPACHE II score of 31-40 while 8.3% had a score of 41-46. A total of 77.7% showed a\nCandida score of 3.Fungal infections are a major cause of morbi-mortality in burned\npatients since they are ideal hosts for opportunistic infections. The incidence of fungal\ninfection was 39.3 for every 1,000 burned patients. On average, the cases of Candidiasis\noccurred after 21.4 days of the admission to the intensive care unit (1-3 weeks).The most\nfrequently isolated yeast was C. albicans in all samples analyzed (wound biopsy samples,\nurocultures, blood cultures, tracheal aspirate cultures, catheter tip cultures). Thus, C.\nalbicans amounted to 53.8%, C. tropicalis to 23.1%, C. parapsilosis sensu stricto\nto13.5%, C. krusei to 5.8%, C. glabrata and C krusei to 1.9 % each. C. albicans was the\nprevalent species in blood cultures, followed by C. parapsilosis. In the urocultures, nonalbicans\nCandida species surpassed C. albicans. Patients with positive blood cultures for\ncandidaemias showed higher mortality than those with negative blood cultures for\ncandidaemias, with no statistically significant difference (p= 0.091). Molecular\nidentification (HWP1 gene amplification by PCR) and microbiological identification agreed with C. albicans and C. dubliniensis. For some C. albicans isolates, the\namplification produced two fragments of DNA. The smaller fragment (941 bp) was\nproduced by the HWP1 gene and the larger fragment resulted from the RBT1 gene partial\namplification by a non-specific primer binding but the identification of C. albicans was\nconfirmed by clone sequencing. Molecular identification of isolates was required for the\nidentification of C. parapsilosis and C. glabrata as C. parapsilosis sensu stricto and C.\nglabrata species complex. For C. glabrata species complex, PCR reaction for RLP31\ngene, which codes for a 60S ribosomal protein, determined the absence of C. bracarensis\nand C. nivariensis, and the presence of C. glabrata. Molecular identification of C.\nparapsilosis species complex by specific primers for each of these complex species by\nsequencing of ITS1 and ITS2 regions of rDNA enabled the identification of C.\nparasilopsis sensu stricto. C. orthosilopsis and C. metapsilosis were not detected
author2	Landaburu, María F.
author_facet	Landaburu, María F. Badino Varela, María Gabriela Mercedes
format	Tesis de maestría Tesis de maestría acceptedVersion
author	Badino Varela, María Gabriela Mercedes
author_sort	Badino Varela, María Gabriela Mercedes
title	Identificación molecular de especies de candida en pacientes quemados
title_short	Identificación molecular de especies de candida en pacientes quemados
title_full	Identificación molecular de especies de candida en pacientes quemados
title_fullStr	Identificación molecular de especies de candida en pacientes quemados
title_full_unstemmed	Identificación molecular de especies de candida en pacientes quemados
title_sort	identificación molecular de especies de candida en pacientes quemados
publisher	Facultad de Farmacia y Bioquímica
publishDate	2017
url	http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_1901 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_1901.dir/1901.PDF
work_keys_str_mv	AT badinovarelamariagabrielamercedes identificacionmoleculardeespeciesdecandidaenpacientesquemados
_version_	1766017487314354176
spelling	I28-R145-HWA_19012019-09-27 Medical histories of 36 burned patients with documented fungal infection,\nadmitted to the Intensive Care Unit of the Hospital Municipal de Quemados del Gobierno\nde la Ciudad de Buenos Aires between January 2011 and December 2014, were analyzed.\nA total of 52 species of Candida were identified by conventional and molecular methods\nfrom the culture collection stored at -20 ºC at the Laboratory of Hospital de Quemados in\nthe patients studied. The mean hospitalization period was 46.7 days, ranging from 9 to\n218 days. They belonged to females 63.9% (n = 23) and 36.1% male (n = 13). The burned\nbody surface area was estimated at 31-50% in 33.3% of the patients (p=0.03). This\npercentage was observed among the higher age group (59 years of age). Instead, patients\nin the group with a mean age of 19 years suffered burns affecting 71% - 100% of their\nbody surface area. With regard to burn depth, fourth-degree burns affected 69.4% of the\npatients, which indicates the seriousness of the injuries in the patients studied. A total of\n75% of the patients showed burns in different locations (torso, face, lower and upper\nlimbs) and only 25% showed injuries only on the lower limbs, upper limbs or torso. The\ncausative agents of burns were: combustible substances (alcohol, gasoline, explosion):\n41.7%, direct fire: 25%, heaters: 13.9%, water: 16.7% and lastly, electricity: 2.8%. It is\nremarkable that in those cases in which the burn causative agent was a combustible\nsubstance, alcohol was the prevalent agent. When studying causative agents in relation to\nage, burn injuries by fire or heaters were found mainly older patients. Conversely, burns\ncaused by alcohol, combustible substances and electricity were reported on younger\npatients. When studying causative agents in relation to sex, alcohol was found to be the\nprevalent element in females. This higher rate for women is associated with the fact that the causative agents of the burn injury are alcohol and direct fire, according to medical\nhistory including injuries by a close third party. They are the current causes of the\nfemicides reported in our country in women under 40 years of age. Electrical burns and\ngasoline burn injuries were affected males exclusively.When analyzing the correlation\nbetween the burned surface area and the cause (combustible substances vs. other agents),\nit was found that causes other than combustible substances generate smaller burn injuries\n(p= 0.01). However, second-degree burns were observed when the agent was a\ncombustible substance (alcohol, gasoline or explosion) (p=0.03), as a result of a deeper\nburn.Fifty per cent of the burned patients showed inhalation injuries (SII), with both\ncombustible substances and fire as causative agents of the burn injury. A central line was\nrequired in 88.9% of the burned patients. Femoral lines were used in 90.6% and upper\nlines (subclavian and/or jugular lines), in 9.4%. The mean number of lines used per\npatient was 5.8. Orotracheal intubation and tracheostomy were performed in 72.2% and\n41.7% of the patients, respectively. A total of 91.7 % of the patients underwent surgical\ntreatment (escharectomy: 100%, escharotomy: 42.4%, fasciotomy: 54.5% and limb\namputation: 6.1%) while 8.3% of the patients did not undergo any surgical treatment\n(only wound lavage). Following surgical treatment and, after reaching viable tissue, the\nwound was covered with skin grafts. Autograft was performed in 97.2% of the patients,\nshowing firm adherence to the underlying tissue in 75% of the cases. In twenty-five per\ncent of the unsuccessful cases, the mortality rate was 100% (p< 0.01), the main cause\nbeing the septic shock. Inotropes were required in 72.2% of the cases and anti-fungal\ndrugs in 91.7%. A total of 58.8% were initially treated with Fluconazole. The rest of the\ncases were treated with Amphotericin B Liposomal. Death occurred in 38.8% of the patients, its rate being higher among patients over 65 years of age (p= 0.018), on\ninotropes (noradrenaline) (p < 0.001), those patients suffering inhalation injury (p= 0.04),\nthose who had undergone fasciotomy (p=0.04) and those with a history or in a high-risk\ngroup (p = 0.002). Infection by bacteria occurred in 100% of the burned patients, the\nmicroorganisms being: Gram-negative bacteria such as multiresistant Acinetobacter\nbaumannii, Pseudomonas aeruginosa and carbapenemase-producing (KPC) Klebsiella\npneumoniae. Among Gram-positive bacteria, the most prevalent was methicillin-resistant\nS. Aureus (MRSA). The mean time of the onset of bacterial infections was 4 days\nfollowing admission to the Intensive Care Unit. A total of 38.9% of the patients had an\nAPACHE II score of 31-40 while 8.3% had a score of 41-46. A total of 77.7% showed a\nCandida score of 3.Fungal infections are a major cause of morbi-mortality in burned\npatients since they are ideal hosts for opportunistic infections. The incidence of fungal\ninfection was 39.3 for every 1,000 burned patients. On average, the cases of Candidiasis\noccurred after 21.4 days of the admission to the intensive care unit (1-3 weeks).The most\nfrequently isolated yeast was C. albicans in all samples analyzed (wound biopsy samples,\nurocultures, blood cultures, tracheal aspirate cultures, catheter tip cultures). Thus, C.\nalbicans amounted to 53.8%, C. tropicalis to 23.1%, C. parapsilosis sensu stricto\nto13.5%, C. krusei to 5.8%, C. glabrata and C krusei to 1.9 % each. C. albicans was the\nprevalent species in blood cultures, followed by C. parapsilosis. In the urocultures, nonalbicans\nCandida species surpassed C. albicans. Patients with positive blood cultures for\ncandidaemias showed higher mortality than those with negative blood cultures for\ncandidaemias, with no statistically significant difference (p= 0.091). Molecular\nidentification (HWP1 gene amplification by PCR) and microbiological identification agreed with C. albicans and C. dubliniensis. For some C. albicans isolates, the\namplification produced two fragments of DNA. The smaller fragment (941 bp) was\nproduced by the HWP1 gene and the larger fragment resulted from the RBT1 gene partial\namplification by a non-specific primer binding but the identification of C. albicans was\nconfirmed by clone sequencing. Molecular identification of isolates was required for the\nidentification of C. parapsilosis and C. glabrata as C. parapsilosis sensu stricto and C.\nglabrata species complex. For C. glabrata species complex, PCR reaction for RLP31\ngene, which codes for a 60S ribosomal protein, determined the absence of C. bracarensis\nand C. nivariensis, and the presence of C. glabrata. Molecular identification of C.\nparapsilosis species complex by specific primers for each of these complex species by\nsequencing of ITS1 and ITS2 regions of rDNA enabled the identification of C.\nparasilopsis sensu stricto. C. orthosilopsis and C. metapsilosis were not detected Fil: Badino Varela, María Gabriela Mercedes. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Buenos Aires, Argentina Landaburu, María F. Facultad de Farmacia y Bioquímica Mujica, María T. Badino Varela, María Gabriela Mercedes 2017-05-18 Se analizaron las historias clínicas de 36 pacientes quemados con infección\nfúngica documentada, internados durante el período comprendido entre enero 2011 y\ndiciembre del 2014 en el área de internación de Terapia Intensiva del Hospital Municipal\nde Quemados del Gobierno de la Ciudad de Buenos Aires. Se identificaron por\nmicrobiología convencional y molecular a 52 especies de Candida provenientes de la\ncolección de cultivo que el Laboratorio del Hospital de Quemados conserva a -20 ºC,\ncorrespondiente a los pacientes estudiados. La internación de los pacientes fue promedio\n46,7 días, con un rango entre 9 y 218 días. Pertenecieron al sexo femenino el 63,9 % (n=23) y al masculino 36,1 % (n=13). Se observó un predominio de una extensión de la\nsuperficie corporal quemada entre 31 y 50 % en el 33,3 % de los pacientes (p=0,03). Esta\nse presentó en el grupo etario de mayor edad (59 años), en tanto entre el 71 y 100%, la\nedad promedio fue de 19 años. Al considerar la profundidad de la quemadura\nespecíficamente la de grado 4, se observó en el 69,4 % de pacientes, esto marca la severa\ngravedad de las lesiones en los pacientes estudiados. El 75 % de los pacientes\npresentaron una combinación de sitios quemados (tronco, cara, miembros inferiores y\nmiembros superiores) y solo el 25 % exhibieron lesiones en miembros inferiores,\nsuperiores o en tronco. Los agentes causantes de la quemadura fueron: combustible\n(alcohol, nafta, explosión) 41,7 %, fuego directo 25,0 %, estufa 13,9 %, agua 16,7 % y\npor último electricidad 2,8 %. Debemos destacar que cuando el agente causal de la\nquemadura resultó el combustible, el alcohol fue el agente de mayor prevalencia. Al\nrelacionar a los agentes causales y la edad observamos que cuando la fuente fue la estufa\ny el fuego las quemaduras se presentaron a una mayor edad promedio, en tanto que\ncuando lo fue el alcohol, combustible y electricidad se presentaron a menor edad. Al\nanalizar los agentes causales y el género, podemos apreciar que el elemento alcohol, fue\nprevalente en el sexo femenino. Esta superioridad femenina, se relaciona con el agente\ncausante alcohol y fuego directo como causa de la quemadura, según figura historia\nclínica por lesiones de un tercero con vínculo afectivo. Son, en parte las causas actuales\nde femicidios, que se informan en nuestro país y que se presentan en menores de 40 años.\nLa quemadura por electricidad y nafta furon exclusivas del sexo masculino.Al relacionar\nla superficie quemada y la causa (combustible vs. otros agentes), se encontró que causas\nno combustibles ocasionan una menor superficie quemada (p= 0,01), en tanto el grado 2 se presenta cuando el agente fue combustible (alcohol, nafta, explosión) (p=0,03), como\nresultado de una mayor profundidad de la quemadura. El 50 % de los pacientes quemados\npresentaron LIH, implicados el combustible y el fuego como agente de la quemadura. Se requirió\nde vías centrales en el de 88,9 % de los quemados. La vía femoral se usó en el 90,6 % y el 9,4 %\nlas vías superiores (subclavia y/o yugular). El promedio de vías utilizadas en cada paciente fue de\n5,8. La intubación orotraqueal y la traqueotomía se practicaron en el 72,2 % y 41,7 %,\nrespectivamente. Se practicó tratamiento quirúrgico en el 91,7 % de los pacientes (escarectomías\n100%, escarotomías 42,4%, fasciotomías 54,5% y amputación de miembros 6,1%) y sin\ntratamiento quirúrgico (sólo curación con lavados) se encontraron 8,3 % de los quemados. Luego\ndel tratamiento quirúrgico y de obtener un lecho de tejido viable, se procedió a cubrir la herida\ncon injertos de piel. Se les aplicó autoinjerto a 97,2%. El mismo exhibió buena adherencia al\ntejido subyacente en un 75%. En el 25% que no fue exitoso, el porcentaje de mortalidad fue de\n100 % (p< 0,01), cuya causa principal es el shock séptico. El requerimiento de inotrópicos fue del\n72,2 % y de antifúngico del 91,7%. Se inició la terapia con fluconazol en el 58,8% y el resto\nanfotericina B liposomal. La mortalidad ocurrió en el 38,8 % pacientes y se observó que fue\nmayor entre los mayores de 65 años (p= 0,018), con el uso de inotrópico (noradrenalina) (p <\n0,001), con LIH (p= 0,04), en pacientes que se les practicó una fasciotomía (p=0.04) y en el grupo\ncon antecedentes o factores de riegos (p = 0,002). Las infecciones bacterianas estuvieron\npresentes en el 100 % de los pacientes quemados y los microorganismos fueron: bacterias Gram\nnegativas multirresistentes como Acinetobacter baumannii, Pseudomonas aeruginosa y\nKlebsiella pneumoniae productora de carbapenemasa (KPC). Entre los Gram positivos, S. aureus\nmeticilinoresistente (SMR) fue el más prevalente. El momento de aparición de las infecciones\nbacterianas en promedio fue 4 días post ingreso a la unidad de cuidados intensivos. El 38,9 % de\nlos pacientes presentaron un APACHE II entre 31-40 y el 8,3 % entre 41?46. El Candida score\nfue de 3 en el 77,7 %. Las infecciones fúngicas son una causa importante de morbimortalidad en pacientes quemados, ya que constituyen un huésped ideal para las infecciones oportunistas. La\nincidencia de infección fúngica representó en 39,3 por cada 1000 quemados y las candidiasis se\nprodujeron en promedio a los 21,4 días de ingreso a la unidad de cuidados intensivos (rango 1-3\nsemanas). La levadura de más frecuente aislamiento fue C. albicans en todos los materiales\nclínicos analizados (biopsia de la lesión, urocultivo, hemocultivo, aspirado traqueal, punta de\ncatéter). Asi, C. albican estuvo representada con el 53,8 %, C. tropicalis con el 23,1 %, C.\nparapsilosis sensu stricto 13,5 %, C. kruse 5,8 %, C. glabrata y C. krusei 1,9 % cada una. En los\nhemocultivos, C. albicans fue la especie prevalente, seguida por C. parapsilosis. En el urocultivo\nlas especies Candida no-C. albicans sobrepasaron a C. albicans. Los pacientes con hemocultivos\npositivos (candidemias), presentaron mayor mortalidad que aquellos con candidemias negativas\naunque sin diferencia estadísticamente significativa (p= 0,091). La identificación molecular (la\namplificación por PCR del gen HWP1 coincidió con la microbiológica en C. albicans y C.\ndubliniensis. Existieron aislamientos de C. albicans que la amplificación produjo dos fragmentos\nde ADN. El fragmento de menor (941 pb) tamaño es producto del gen HWP1 y el fragmento de\nmayor tamaño se debió a la amplificación parcial del gen RBT1 por un pegado inespecífico de los\ncebadores, pero se confirmó por secuenciación de ambos clones a C. albicans.Para la\nidentificación de C. parapsilosis y de C. glabrata como C. parapsilosis sensu stricto y C.\nglabrata dentro del complejo de esta última especie, se requirió de la identificación molecular a\nlos aislamientos. En el complejo de C. glabrata a través de la reacción de PCR para el gen RLP31\nque codifica para una proteína ribosomal de 60S determinó la ausencia de C. bracarensis y C.\nnivariensis; y confirmó la presencia de C. glabrata. La identificación molecular de las especies\ndel complejo de C. parapsilosis con cebadores específicos para cada una de las especies de este\ncomplejo derivados de secuencias dentro de la región de ITS1 y ITS2 del rDNA permitió la\nidentificación C. parasilopsi sensu stricto. No se detectó la presencia de C. orthosilopsis y C.\nmetapsilosis.\n application/pdf Catalano, Mariana Iovannitti, Cristina Arechavala, Alicia Candida Quemaduras Infecciones fúngicas Patogénesis de quemaduras spa Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/2.5/ar/ Ciencias de la vida Identificación molecular de especies de candida en pacientes quemados info:eu-repo/semantics/masterThesis info:ar-repo/semantics/tesis de maestría info:eu-repo/semantics/acceptedVersion http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_1901 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_1901.dir/1901.PDF

Identificación molecular de especies de candida en pacientes quemados

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