Identificación fenotípica y molecular de Coccidioides posadasii en una paciente examinada para un trasplante bipulmonar / Phenotypic and molecular identification of Coccidioides posadasii in a patient evaluated for bilateral lung transplantation

Antecedentes. La coccidioidomicosis es una infección micótica endémica debida a Coccidioides immitis y Coccidioides posadasii. Su presencia en receptores de un trasplante puede revestir especial gravedad, en particular en pacientes con una infección activa o previa por Coccidioides. En este grupo de pacientes se han descrito casos letales de la infección. Objetivos. Describimos un caso grave de neumonía debida a C. posadassi en una mujer blanca de 29 años de edad, que fue ingresada en el hospital como parte de la valoración de un trasplante bipulmonar. La paciente había nacido y residía en Catamarca, Argentina. La metodología molecular contribuyó a la identificación de la especie. Métodos. Para establecer el diagnóstico de la infección e identificar C. posadasii, se efectuaron una exploración clínica, pruebas de laboratorio y exámenes microbiológicos. Resultados. A partir del cultivo del líquido de lavado broncoalveolar se aisló el hongo. La caracterización fenotípica, la reacción en cadena de la polimerasa específica y la inoculación experimental a un animal demostraron la presencia de C. posadasii. La paciente respondió al tratamiento con anfotericina B desoxicolato. El trasplante de pulmón se difirió. Conclusiones. La RCP específica puede ser una importante alternativa para la correcta identificación de C. immitis o C. posadasii en los laboratorios con medios implementados de biología molecular. El caso descrito destaca la necesidad de que, en las unidades de trasplante de órganos, se efectúe una valoración sistemática de los pacientes que viven en áreas endémicas de coccidioidomicosis(AU)

Background. Coccidioidomycosis is an endemic fungal infection caused by Coccidioides immitis and Coccidioides posadasii. It can be particularly severe in transplant recipients that have a current or a previous coccidioidal infection. Fatal case of coccidioidomycosis has been described in this group of patients. Aims. We report a severe case of pneumonia caused by C. posadassi in a 29 year-old white woman that had been admitted to hospital as part of the evaluation for bilateral lung transplantation. The patient was a native and resident of Catamarca, Argentina. Molecular methodologies contributed to the species identification. Methods. Clinical, laboratory records and microbiological tests were carried out to diagnose the infection and to identify C. posadasii. Results. A fungus was isolated from BAL culture. Phenotypic characterization, specific PCR and experimental animal inoculation demonstrated the presence of C. posadasii. The patient responded well to amphotericin B deoxycholate. Lung transplantation was postponed. Conclusions. Specific PCR can be an important alternative for the correct identification of C. immitis or C. posadasii in laboratories with implemented molecular biology tools. This case emphasizes the need for a systematic assessment in organ transplant units of patients inhabiting endemic areas of coccidioidomycosis(AU)

Phenotypic and molecular identification of Coccidioides posadasii in a patient evaluated for bilateral lung transplantation.

BACKGROUND: Coccidioidomycosis is an endemic fungal infection caused by Coccidioides immitis and Coccidioides posadasii. It can be particularly severe in transplant recipients that have a current or a previous coccidioidal infection. Fatal case of coccidioidomycosis has been described in this group of patients. AIMS: We report a severe case of pneumonia caused by C. posadassi in a 29 year-old white woman that had been admitted to hospital as part of the evaluation for bilateral lung transplantation. The patient was a native and resident of Catamarca, Argentina. Molecular methodologies contributed to the species identification. METHODS: Clinical, laboratory records and microbiological tests were carried out to diagnose the infection and to identify C. posadasii. RESULTS: A fungus was isolated from BAL culture. Phenotypic characterization, specific PCR and experimental animal inoculation demonstrated the presence of C. posadasii. The patient responded well to amphotericin B deoxycholate. Lung transplantation was postponed. CONCLUSIONS: Specific PCR can be an important alternative for the correct identification of C. immitis or C. posadasii in laboratories with implemented molecular biology tools. This case emphasizes the need for a systematic assessment in organ transplant units of patients inhabiting endemic areas of coccidioidomycosis.

Capacidad de los laboratorios nacionales de referencia en Latinoamérica para detectar mecanismos de resistencia emergentes / Capability of national reference laboratories in Latin America to detect emerging resistance mechanisms

Objetivo. Evaluar la capacidad de 17 laboratorios nacionales de referencia que participan en el Programa Latinoamericano de Control de Calidad en Bacteriología y Resistencia a los Antimicrobianos (LA-EQAS) para detectar mecanismos de resistencia emergentes, a saber: resistencia de enterobacterias a carbapenemes por presencia de Klebsiella pneumoniae carbapenemasa (KPC); resistencia de enterobacterias a carbapenemes por presencia de metalobetalactamasas (MBL) tipo IMP, y resistencia intermedia a vancomicina de aislamientos de Staphylococcus aureus (VISA). Métodos. Se enviaron los siguientes tres aislamientos a los 17 laboratorios participantes del LA-EQAS: Klebsiella pneumoniae OPS-161 productor de KPC, Enterobacter cloacae OPS-166 productor de IMP y S. aureus OPS-165 con resistencia intermedia a vancomicina. Se evaluó la interpretación de las pruebas de sensibilidad y detección del mecanismo de resistencia y el tamaño de los halos de inhibición (método de difusión por discos) o valor de la concentración inhibitoria mínima (CIM). Resultados. La concordancia en la detección de los mecanismos de resistencia fue de 76,4%, 73,3% y 66,7% con respecto a la cepas K. pneumoniae OPS-161, E. cloacae OPS-166 y S. aureus OPS-165, respectivamente. La concordancia entre las zonas de inhibición obtenidas por los laboratorios participantes y los rangos establecidos por el laboratorio coordinador fue aceptable en los tres aislamientos, ya que alcanzó 90,8%, 92,8% y 88,9%, respectivamente, para cada cepa. Conclusiones. La concordancia global en la detección de los mecanismos de resistencia KPC, MBL y VISA fue de 72,1%. Consideramos que los laboratorios nacionales de referencia de América Latina son capaces de reconocer estos mecanismos de resistencia emergentes y se espera que en el futuro la concordancia alcance su nivel máximo.

Objective. To evaluate the capability of 17 national reference laboratories participating in the Latin American Quality Control Program in Bacteriology and Antibiotic Resistance (LA-EQAS) to detect emerging resistance mechanisms— namely: resistance of enterobacteria to carbapenems due to the presence of Klebsiella pneumoniae carbapenemase (KPC) and metallo-beta-lactamase (MBL) type IMP, and intermediate resistance of Staphylococcus aureus isolates to vancomycin (vancomycinintermediate resistant S. aureus—VISA). Methods. The following three isolates were sent to the 17 participating LA-EQAS laboratories: KPC-producing Klebsiella pneumoniae PAHO-161, IMP-producing Enterobacter cloacae PAHO-166, and S. aureus PAHO-165 with intermediate resistance to vancomycin. Performance of each of the following operations was evaluated: interpretation of sensitivity tests, detection of the resistance mechanism, and assessment of either inhibition halo size (disk diffusion method) or minimum inhibitory concentration (MIC). Results. Concordance in the detection of resistance mechanisms was 76.4%, 73.3%, and 66.7% for the K. pneumoniae PAHO-161, E. cloacae PAHO-166, and S. aureus PAHO- 165 strains, respectively. Concordance between the inhibition areas observed by the participating laboratories and the ranges established by the coordinating laboratory was acceptable for all three isolates, at 90.8%, 92.8%, and 88.9%, respectively. Conclusions. Overall concordance in on the detection of KPC, MBL, and VISA resistance mechanisms was 72.1%. We consider the national reference laboratories in Latin America capable of recognizing these emerging resistance mechanisms and expect that maximum levels of concordance will be reached in the future.

[Capability of national reference laboratories in Latin America to detect emerging resistance mechanisms]. / Capacidad de los laboratorios nacionales de referencia en Latinoamérica para detectar mecanismos de resistencia emergentes.

OBJECTIVE: To evaluate the capability of 17 national reference laboratories participating in the Latin American Quality Control Program in Bacteriology and Antibiotic Resistance (LA-EQAS) to detect emerging resistance mechanisms- namely: resistance of enterobacteria to carbapenems due to the presence of Klebsiella pneumoniae carbapenemase (KPC) and metallo-beta-lactamase (MBL) type IMP, and intermediate resistance of Staphylococcus aureus isolates to vancomycin (vancomycin-intermediate resistant S. aureus-VISA). METHODS: The following three isolates were sent to the 17 participating LA-EQAS laboratories: KPC -producing Klebsiella pneumoniae PAHO-161, IMP-producing Enterobacter cloacae PAHO-166, and S. aureus PAHO-165 with intermediate resistance to vancomycin. Performance of each of the following operations was evaluated: interpretation of sensitivity tests, detection of the resistance mechanism, and assessment of either inhibition halo size (disk diffusion method) or minimum inhibitory concentration (MIC). RESULTS: Concordance in the detection of resistance mechanisms was 76.4%, 73.3%, and 66.7% for the K. pneumoniae PAHO-161, E. cloacae PAHO-166, and S. aureus PAHO-165 strains, respectively. Concordance between the inhibition areas observed by the participating laboratories and the ranges established by the coordinating laboratory was acceptable for all three isolates, at 90.8%, 92.8%, and 88.9%, respectively. CONCLUSIONS: Overall concordance in on the detection of KPC, MBL, and VISA resistance mechanisms was 72.1%. We consider the national reference laboratories in Latin America capable of recognizing these emerging resistance mechanisms and expect that maximum levels of concordance will be reached in the future.

Complex class 1 integrons with diverse variable regions, including aac(6')-Ib-cr, and a novel allele, qnrB10, associated with ISCR1 in clinical enterobacterial isolates from Argentina.

Transferable quinolone resistance has not previously been reported in Argentina. Here we describe three complex class 1 integrons harboring the novel allele qnrB10 in a unique region downstream of orf513, one of them also containing aac(6')-Ib-cr within the variable region of integrons. The three arrays differed from bla(CTX-M-2)-bearing integrons, which are broadly distributed in Argentina.

Molecular characterization of methicillin-resistant Staphylococcus aureus isolates reveals the spreading of a new clone in Buenos Aires City.

We performed a hospital-acquired methicillin-resistant Staphylococcus aureus molecular study in Buenos Aires city. Four clones were found harboring the transposable elements Tn4001 and Tn5405 and the erythromycin resistance determinants ermA and mef(E). Of the isolates, 73% belonged to a clone found previously in the city of Córdoba, which showed an epidemic behavior initially attributed to the widely disseminated South American clone.

Molecular characterization of methicillin-resistant staphylococcus aureus isolates reveals the spreading of a new clone in Buenos Aires city

Realizamos un estudio molecular en aislamientos de Staphylococcus aureus adquiridos en el ambiente hospitalario de la ciudad de Buenos Aires. Se hallaron cuatro clones que albergaban los elementos transponibles Tn4001 y Tn5405 y los determinantes de resistencia a eritromicina, ermA y mef(E). El 73% de los aislamientos pertenecían a un clon hallado previamente en la ciudad de Córdoba con características epidemiológicas atribuidas inicialmente al clon sudamericano ampliamente diseminado en el mundo.

Molecular characterization of methicillin-resistant staphylococcus aureus isolates reveals the spreading of a new clone in Buenos Aires city

Realizamos un estudio molecular en aislamientos de Staphylococcus aureus adquiridos en el ambiente hospitalario de la ciudad de Buenos Aires. Se hallaron cuatro clones que albergaban los elementos transponibles Tn4001 y Tn5405 y los determinantes de resistencia a eritromicina, ermA y mef(E). El 73% de los aislamientos pertenecían a un clon hallado previamente en la ciudad de Córdoba con características epidemiológicas atribuidas inicialmente al clon sudamericano ampliamente diseminado en el mundo. (AU)

Molecular characterization of methicillin-resistant staphylococcus aureus isolates reveals the spreading of a new clone in Buenos Aires city

Realizamos un estudio molecular en aislamientos de Staphylococcus aureus adquiridos en el ambiente hospitalario de la ciudad de Buenos Aires. Se hallaron cuatro clones que albergaban los elementos transponibles Tn4001 y Tn5405 y los determinantes de resistencia a eritromicina, ermA y mef(E). El 73% de los aislamientos pertenecían a un clon hallado previamente en la ciudad de Córdoba con características epidemiológicas atribuidas inicialmente al clon sudamericano ampliamente diseminado en el mundo. (AU)

Emergence of a Streptococcus pneumoniae clinical isolate highly resistant to telithromycin and fluoroquinolones.

Streptococcus pneumoniae is a major pathogen causing community-acquired pneumonia and acute bronchitis. Macrolides, fluoroquinolones (FQs), and, recently, telithromycin (TEL) constitute primary therapeutic options, and rare cases of resistance have been reported. In this report, we describe the emergence of an S. pneumoniae clinical isolate with high-level TEL resistance (MIC, 256 microg/ml) and simultaneous resistance to FQs. Ongoing studies are oriented to elucidate the precise mechanism of resistance to TEL.

Erythromycin-resistant Streptococcus pyogenes in Argentina.

Erythromycin (ERY) resistance in Streptococcus pyogenes has recently emerged as a problem of growing concern all through the world. We are presenting the comparison of results of the continuous surveillance of erythromycin resistance in S. pyogenes performed since 1989 in the Hospital de Pediatría J.P. Garrahan of Buenos Aires City, with independently observed rates in other five centers of Buenos Aires and seven centers of six other Argentinian cities, obtained between 1999 and 2001. A significant increase of erythromycin resistance was observed among S. pyogenes isolated in the Hospital Garrahan (6.6% in 1998-1999 to 9.9% in 2000). Similar trends were also detected in other centers of other Argentinian cities when recent data were compared to results of a multicenter study performed in 1995. However, lower rates of resistance were recorded in Mendoza, Cipolletti and Neuquén in comparison with data of 1995, 1998 and 1998 respectively. The reason of such decreasing resistance rates deserves to be investigated. The average of ERY-resistance rates obtained in the surveyed centers was 6.7% (range 0.5-14.1%). Control of antimicrobial use should be performed to warrant the future effectiveness of macrolide antibiotics regarding the positive association between use and resistance. These results also suggest that susceptibility tests for macrolides should be performed whenever S. pyogenes is isolated in Argentina.

Erythromycin-resistant Streptococcus pyogenes in Argentina.

Erythromycin (ERY) resistance in Streptococcus pyogenes has recently emerged as a problem of growing concern all through the world. We are presenting the comparison of results of the continuous surveillance of erythromycin resistance in S. pyogenes performed since 1989 in the Hospital de Pediatría J.P.Garrahan of Buenos Aires City, with independently observed rates in other five centers of Buenos Aires and seven centers of six other Argentinian cities, obtained between 1999 and 2001. A significant increase of erythromycin resistance was observed among S. pyogenes isolated in the Hospital Garrahan (6.6

in 1998-1999 to 9.9

in 2000). Similar trends were also detected in other centers of other Argentinian cities when recent data were compared to results of a multicenter study performed in 1995. However, lower rates of resistance were recorded in Mendoza, Cipolletti and Neuquén in comparison with data of 1995, 1998 and 1998 respectively. The reason of such decreasing resistance rates deserves to be investigated. The average of ERY-resistance rates obtained in the surveyed centers was 6.7

(range 0.5-14.1

). Control of antimicrobial use should be performed to warrant the future effectiveness of macrolide antibiotics regarding the positive association between use and resistance. These results also suggest that susceptibility tests for macrolides should be performed whenever S. pyogenes is isolated in Argentina.

Conservación de catéteres a 4 °C. Influencia sobre los cultivos cuantitativos / Catheter storage at 4 ºC. Influence on quantitative cultures

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El laboratorio de microbiología en el diagnóstico de la bacteriemia relacionada con los catéteres / The clinical microbiology laboratory in the diagnosis of catheter related bacteremia

La sepsis relacionada con los catéteres es un problema de gran importancia en pacientes de todas las edades. El diagnóstico microbiológico debe contemplar las principales vías de infección (interfase piel-catéter y la vía endoluminal). Con esta finalidad, analizamos 1.496 catéteres centrales y periféricos de corta permanencia y 119 episodios de bacteriemia asociada con catéteres (BAC). Éstos fueron sembrados por la técnica cuantitativa de Brun Buisson (QT), la semicuantitativa de Maki (SQ) y la cualitativa (QL) en caldo, obteniéndose los siguientes valores de sensibilidad, especificidad, valor predictivo positivo, valor predictivo negativo e índice de Youden: SQ = 87 por ciento; 88 por ciento; 40 por ciento; 99 por ciento; 0,75; QT (>= 100 unidades formadoras de colonias [UFC]/ml) = 88 por ciento, 89 por ciento, 43 por ciento, 99 por ciento, 0,77; QT (>= 1.000 UFC/ml) = 77 por ciento, 92 por ciento, 48 por ciento, 97 por ciento, 0,69, y QL = 94 por ciento, 68 por ciento, 20 por ciento, 99 por ciento, 0,62. Los resultados entre SQ y QT (>= 100 UFC/ml) fueron comparables, y el agregado de QT a SQ permitió incrementar la detección de BAC en el 12,8 por ciento, en tanto que la situación inversa se dio en el 10 por ciento de los casos. De acuerdo con estos datos es aconsejable combinar rutinariamente SQ y QT. Finalmente, en 42 episodios de BAC asociados a catéteres implantables procesados por medio del cultivo cuantitativo diferencial de sangre obtenida a través de catéter frente a sangre obtenida de vena periférica, el 79 por ciento tuvieron una relación superior a 1.000, el 7 por ciento entre 100 y 1.000, el 9 por ciento entre 10 y 100 y sólo el 5 por ciento entre 5 y 10 UFC/ml (AU)