Utility evaluation of two molecular methods for Leptospira spp. typing in human serum samples.

Most of the available genotyping methods were applied and evaluated in Leptospira isolates and only few of them in a relevant sample size of blood specimens but not of sera. The objective of this study was to evaluate the utility of one partial 16S rRNA gene sequencing assay (16S rRNA) and an optimized. Multilocus sequence typing scheme (MLST) for Leptospira typing directly in serum samples. Confirmed leptospirosis patients (n = 228) from Argentina (2005-2016) were randomly included. Septicemic-phase serum samples (n = 228) were studied by two genotyping methods. Available immune-phase serum samples of the included patients (n = 159) were studied by MAT to compare serological and molecular results. In culture-proven cases (n = 8), genotyping results between clinical samples and isolates were compared. Typing success rate (TSR) was 21.9% for 16S rRNA and 11.4% for MLST (full allelic profile) and a positive trend in both TSR during the study period was observed. Two species (L. interrogans and L. borgpertesenii) were identified by both methods and MLST assigned 8 different STs. The probable serogroups identified by MLST were coincident with the presumptive infecting serogroups identified by MAT, but with different frequencies. The three serogroups (Canicola, Sejroe and Icterohaemorrhagiae) most frequently identified by MAT were also genotyped by MLST. Typing results via 16S rRNA and MLST in clinical samples and isolates of culture-proven cases, were consistent except for one case. Performance of partial 16S rRNA gene sequencing assay and the optimized MLST scheme directly in sera may increase and improve the knowledge about species and serogroups causing human leptospirosis, especially in countries with low rates of culture sample collection or Leptospira isolation.

Primer informe de Leptospira interrogans en el roedor sigmodontino Scapteromys aquaticus

[RESUMEN]. La leptospirosis es una enfermedad zoonótica de distribución mundial que puede transmitirse por contacto directo o indirecto con orina o tejidos de animales infectados. En Argentina, la leptospirosis es endémica en la provincia de Santa Fe y presenta brotes epidémicos durante las inundaciones. Sin embargo, se sabe muy poco sobre el papel que cumplen los roedores silvestres en la diseminación de la enfermedad en el país. El objetivo de este estudio fue identificar las especies hospederas de leptospiras patógenas entre los roedores presentes en un asentamiento ribereño de la provincia de Santa Fe. Se realizó un muestreo de roedores durante octubre de 2015. Los riñones de los animales capturados se analizaron por real-time PCR para el gen LipL32 de leptospiras patógenas. En los animales que resultaron positivos, se realizó test de microaglutinación (MAT) y tipificación molecular por amplificación del gen 16S rRNA y dos esquemas de MLST. Se capturaron 37 roedores de las especies Akodon azarae, Cavia aperea, Oligoryzomys flavescens, Rattus rattus y Scapteromys aquaticus. En el análisis por real-time PCR resultó positivo un macho de Scapteromys aquaticus. El suero de este individuo y del resto de los S. aquaticus capturados (n = 18) se analizaron por test de microaglutinación (MAT), y fueron no reactivos para los 10 serovares probados. La amplificación del gen 16S rRNA identificó la especie infectante como Leptospira interrogans, mientras que no se obtuvo amplificación para los dos esquemas de MLST. El hallazgo de este estudio aporta nueva información acerca de presencia de leptospiras patógenas en roedores silvestres, que es relevante para la zona por tratarse de una especie ampliamente distribuida en ambientes pantanosos e inundables de América del Sur.

[ABSTRACT]. Leptospirosis is a globally distributed zoonosis that can be transmitted through direct or indirect contact with the urine or tissues of infected animals. In Argentina, leptospirosis is endemic in the province of Santa Fe and epidemic outbreaks occur during floods. However, very little is known about the role that wild rodents play in the spread of the disease in Argentina. The objective of this study was to identify the host species of pathogenic Leptospira among rodents in a riverine settlement in the province of Santa Fe. We conducted a trapping session in October 2015. Kidneys of the captured animals were analyzed by real-time PCR for the LipL32 gene of pathogenic Leptospira. Animals that were positive were subjected to microscopic agglutination test (MAT) and molecular typing by amplification of the 16S rRNA gene and two multilocus sequence typing (MLST) schemes. A total of 37 rodents of the species Akodon azarae, Cavia aperea, Oligoryzomys flavescens, Rattus rattus, and Scapteromys aquaticus were captured. Real-time PCR found one male Scapteromys aquaticus that was positive. The serum of this individual and of the rest of the S. aquaticus captured (n = 18) were analyzed by MAT and were non-reactive for the 10 serovars tested. Amplification of the 16S rRNA gene identified the infective species as Leptospira interrogans, while amplification could not be obtained for the two MLST schemes. The findings of this study contribute new information concerning the presence of pathogenic Leptospira in wild rodents, which is relevant in this region because the species is widely distributed in swampy and flood-prone environments of South America.

[RESUMO]. A leptospirose é uma doença zoonótica de distribuição mundial transmitida pelo contato direto ou indireto com a urina ou os tecidos de animais infectados. Na Argentina, a leptospirose é endêmica na Província de Santa Fé com surtos epidêmicos ocorrendo com as enchentes. Sabe-se pouco sobre o papel dos roedores silvestres na propagação da doença no país. O objetivo deste estudo foi identificar as espécies hospedeiras de leptospiras patogênicas em roedores encontrados em um núcleo de povoamento ribeirinho na Província de Santa Fé. A amostragem dos roedores foi feita no mês de outubro de 2015. Os tecidos dos rins dos animais capturados foram analisados com a técnica de reação em cadeia da polimerase em tempo real (PCR-RT) quanto à presença do gene LipL32 de leptospiras patógenas. Para os animais com resultados positivos, foi realizado o teste de microaglutinação (MAT) e tipagem molecular baseada na amplificação do gene 16S rRNA e dois esquemas de tipagem por sequenciamento de locos múltiplos (MLST). Ao todo, foram capturados 37 roedores das espécies Akodon azarae, Cavia aperea, Oligoryzomys flavescens, Rattus e Scapteromys aquaticus. O ensaio de PCR-RT foi positivo em um roedor macho da espécie Scapteromys aquaticus. Os soros deste animal e dos outros S. aquaticus capturados (n = 18) foram analisados com o MAT e os resultados foram não reagentes para os 10 sorovares testados. A amplificação do gene 16S rRNA permitiu identificar a espécie infetante como sendo Leptospira interrogans e não houve amplificação nos dois esquemas de MLST. O achado deste estudo fornece um novo dado quanto à presença de leptospiras patogênicas em roedores silvestres, importante para esta área por se tratar de uma espécie de ampla distribuição em terras pantanosas e inundáveis da América do Sul.

[First report on Leptospira interrogans in the sigmodontine rodent Scapteromys aquaticusPrimeiro relato da presença de Leptospira interrogans em roedores sigmodontíneos Scapteromys aquaticus]. / Primer informe de Leptospira interrogans en el roedor sigmodontino Scapteromys aquaticus.

Leptospirosis is a globally distributed zoonosis that can be transmitted through direct or indirect contact with the urine or tissues of infected animals. In Argentina, leptospirosis is endemic in the province of Santa Fe and epidemic outbreaks occur during floods. However, very little is known about the role that wild rodents play in the spread of the disease in Argentina. The objective of this study was to identify the host species of pathogenic Leptospira among rodents in a riverine settlement in the province of Santa Fe.We conducted a trapping session in October 2015. Kidneys of the captured animals were analyzed by real-time PCR for the LipL32 gene of pathogenic Leptospira. Animals that were positive were subjected to microscopic agglutination test (MAT) and molecular typing by amplification of the 16S rRNA gene and two multilocus sequence typing (MLST) schemes.A total of 37 rodents of the species Akodon azarae, Cavia aperea, Oligoryzomys flavescens, Rattus rattus, and Scapteromys aquaticus were captured. Real-time PCR found one male Scapteromys aquaticus that was positive. The serum of this individual and of the rest of the S. aquaticus captured (n = 18) were analyzed by MAT and were non-reactive for the 10 serovars tested. Amplification of the 16S rRNA gene identified the infective species as Leptospira interrogans, while amplification could not be obtained for the two MLST schemes.The findings of this study contribute new information concerning the presence of pathogenic Leptospira in wild rodents, which is relevant in this region because the species is widely distributed in swampy and flood-prone environments of South America.

A leptospirose é uma doença zoonótica de distribuição mundial transmitida pelo contato direto ou indireto com a urina ou os tecidos de animais infectados. Na Argentina, a leptospirose é endêmica na Província de Santa Fé com surtos epidêmicos ocorrendo com as enchentes. Sabe-se pouco sobre o papel dos roedores silvestres na propagação da doença no país. O objetivo deste estudo foi identificar as espécies hospedeiras de leptospiras patogênicas em roedores encontrados em um núcleo de povoamento ribeirinho na Província de Santa Fé.A amostragem dos roedores foi feita no mês de outubro de 2015. Os tecidos dos rins dos animais capturados foram analisados com a técnica de reação em cadeia da polimerase em tempo real (PCR-RT) quanto à presença do gene LipL32 de leptospiras patógenas. Para os animais com resultados positivos, foi realizado o teste de microaglutinação (MAT) e tipagem molecular baseada na amplificação do gene 16S rRNA e dois esquemas de tipagem por sequenciamento de locos múltiplos (MLST).Ao todo, foram capturados 37 roedores das espécies Akodon azarae, Cavia aperea, Oligoryzomys flavescens, Rattus e Scapteromys aquaticus. O ensaio de PCR-RT foi positivo em um roedor macho da espécie Scapteromys aquaticus. Os soros deste animal e dos outros S. aquaticus capturados (n = 18) foram analisados com o MAT e os resultados foram não reagentes para os 10 sorovares testados. A amplificação do gene 16S rRNA permitiu identificar a espécie infetante como sendo Leptospira interrogans e não houve amplificação nos dois esquemas de MLST.O achado deste estudo fornece um novo dado quanto à presença de leptospiras patogênicas em roedores silvestres, importante para esta área por se tratar de uma espécie de ampla distribuição em terras pantanosas e inundáveis da América do Sul.

Primer informe de Leptospira interrogans en el roedor sigmodontino Scapteromys aquaticus / First report on Leptospira interrogans in the sigmodontine rodent Scapteromys aquaticus / Primeiro relato da presença de Leptospira interrogans em roedores sigmodontíneos Scapteromys aquaticus

RESUMEN La leptospirosis es una enfermedad zoonótica de distribución mundial que puede transmitirse por contacto directo o indirecto con orina o tejidos de animales infectados. En Argentina, la leptospirosis es endémica en la provincia de Santa Fe y presenta brotes epidémicos durante las inundaciones. Sin embargo, se sabe muy poco sobre el papel que cumplen los roedores silvestres en la diseminación de la enfermedad en el país. El objetivo de este estudio fue identificar las especies hospederas de leptospiras patógenas entre los roedores presentes en un asentamiento ribereño de la provincia de Santa Fe. Se realizó un muestreo de roedores durante octubre de 2015. Los riñones de los animales capturados se analizaron por real-time PCR para el gen LipL32 de leptospiras patógenas. En los animales que resultaron positivos, se realizó test de microaglutinación (MAT) y tipificación molecular por amplificación del gen 16S rRNA y dos esquemas de MLST. Se capturaron 37 roedores de las especies Akodon azarae, Cavia aperea, Oligoryzomys flavescens, Rattus rattus y Scapteromys aquaticus. En el análisis por real-time PCR resultó positivo un macho de Scapteromys aquaticus. El suero de este individuo y del resto de los S. aquaticus capturados (n = 18) se analizaron por test de microaglutinación (MAT), y fueron no reactivos para los 10 serovares probados. La amplificación del gen 16S rRNA identificó la especie infectante como Leptospira interrogans, mientras que no se obtuvo amplificación para los dos esquemas de MLST. El hallazgo de este estudio aporta nueva información acerca de presencia de leptospiras patógenas en roedores silvestres, que es relevante para la zona por tratarse de una especie ampliamente distribuida en ambientes pantanosos e inundables de América del Sur.(AU)

ABSTRACT Leptospirosis is a globally distributed zoonosis that can be transmitted through direct or indirect contact with the urine or tissues of infected animals. In Argentina, leptospirosis is endemic in the province of Santa Fe and epidemic outbreaks occur during floods. However, very little is known about the role that wild rodents play in the spread of the disease in Argentina. The objective of this study was to identify the host species of pathogenic Leptospira among rodents in a riverine settlement in the province of Santa Fe. We conducted a trapping session in October 2015. Kidneys of the captured animals were analyzed by real-time PCR for the LipL32 gene of pathogenic Leptospira. Animals that were positive were subjected to microscopic agglutination test (MAT) and molecular typing by amplification of the 16S rRNA gene and two multilocus sequence typing (MLST) schemes. A total of 37 rodents of the species Akodon azarae, Cavia aperea, Oligoryzomys flavescens, Rattus rattus, and Scapteromys aquaticus were captured. Real-time PCR found one male Scapteromys aquaticus that was positive. The serum of this individual and of the rest of the S. aquaticus captured (n = 18) were analyzed by MAT and were non-reactive for the 10 serovars tested. Amplification of the 16S rRNA gene identified the infective species as Leptospira interrogans, while amplification could not be obtained for the two MLST schemes. The findings of this study contribute new information concerning the presence of pathogenic Leptospira in wild rodents, which is relevant in this region because the species is widely distributed in swampy and flood-prone environments of South America.(AU)

RESUMO A leptospirose é uma doença zoonótica de distribuição mundial transmitida pelo contato direto ou indireto com a urina ou os tecidos de animais infectados. Na Argentina, a leptospirose é endêmica na Província de Santa Fé com surtos epidêmicos ocorrendo com as enchentes. Sabe-se pouco sobre o papel dos roedores silvestres na propagação da doença no país. O objetivo deste estudo foi identificar as espécies hospedeiras de leptospiras patogênicas em roedores encontrados em um núcleo de povoamento ribeirinho na Província de Santa Fé. A amostragem dos roedores foi feita no mês de outubro de 2015. Os tecidos dos rins dos animais capturados foram analisados com a técnica de reação em cadeia da polimerase em tempo real (PCR-RT) quanto à presença do gene LipL32 de leptospiras patógenas. Para os animais com resultados positivos, foi realizado o teste de microaglutinação (MAT) e tipagem molecular baseada na amplificação do gene 16S rRNA e dois esquemas de tipagem por sequenciamento de locos múltiplos (MLST). Ao todo, foram capturados 37 roedores das espécies Akodon azarae, Cavia aperea, Oligoryzomys flavescens, Rattus e Scapteromys aquaticus. O ensaio de PCR-RT foi positivo em um roedor macho da espécie Scapteromys aquaticus. Os soros deste animal e dos outros S. aquaticus capturados (n = 18) foram analisados com o MAT e os resultados foram não reagentes para os 10 sorovares testados. A amplificação do gene 16S rRNA permitiu identificar a espécie infetante como sendo Leptospira interrogans e não houve amplificação nos dois esquemas de MLST. O achado deste estudo fornece um novo dado quanto à presença de leptospiras patogênicas em roedores silvestres, importante para esta área por se tratar de uma espécie de ampla distribuição em terras pantanosas e inundáveis da América do Sul.(AU)

Leptospirosis humana en Argentina: un esquema de análisis, 2014 / Human Leptospirosis in Argentina: An Analysis Scheme, 2014

INTRODUCCIÓN: En general, sólo existen estimaciones sobre el número de casos de leptospirosis en las Américas. La fuente más común son los Ministerios de Salud, tanto nacionales como provinciales, que proporcionan datos útiles sobre tendencias de incidencia de la leptospirosis, identificación de brotes y efectos de intervenciones gubernamentales. OBJETIVOS: Conocer la incidencia de casos de leptospirosis detectados por laboratorio durante 2014 en Argentina y generar un esquema de análisis de bases de datos de diferentes organismos nacionales para que sea repetido y difundido anualmente. MÉTODOS: Se analizaron las bases de datos del Sistema de Vigilancia Laboratorial (SIVILA)y del Instituto Nacional de Enfermedades Respiratorias (INER), teniendo en cuenta los casos notificados en 2014, según la fecha de inicio de síntomas. RESULTADOS: La incidencia de casos confirmados de leptospirosis en 2014 fue de 0,32/100 000 habitantes, mayormente detectados en las provincias de Santa Fe, Buenos Aires y Entre Ríos. Los serogrupos circulantes más prevalentes fueron Icterohaemorrhagiae, Sejroe y Canicola. CONCLUSIONES: Se resalta la utilidad de este tipo de estudios, que incentivan la búsqueda de casos, notificación y vigilancia de la leptospirosis, tanto para mejorar el conocimiento de la incidencia de la enfermedad y su distribución como para orientar las acciones hacia los lugares de mayor riesgo del país.

INTRODUCTION: In general, there are only estimates of the number of cases in the Americas. This data is commonly obtained by both national and provincial Ministries of Health which, despite data limitations, provide useful information on leptospirosis incidence trends, outbreak identification and effects of government interventions. OBJECTIVES: To know the incidence of leptospirosis cases detected by laboratory during 2014 in Argentina, and to generate an analysis scheme of databases from different national organisms to be repeated and diffused annually. METHODS: The SIVILA and INER databases were analyzed, taking into account the cases reported in 2014, according to the date of onset of symptoms. RESULTS: The incidence of confirmed leptospirosis cases in 2014 was 0.32/100 000 persons, mostly detected in Santa Fe, Buenos Aires and Entre Ríos provinces. The most prevalent circulating serogroups were Icterohaemorrhagiae, Sejroe and Canicola. CONCLUSIONS: These studies motivate the suspicion, notification and surveillance of leptospirosis and are useful both to improve the knowledge of the incidence of cases and their distribution, as well as to guide actions towards the most risky places in the country.

Ampliación del sistema de Vigilancia intensificada de leptospirosis en las provincias de Santa Fe, Entre Ríos y Buenos Aires (año 2016) / Expansion of the intensified leptospirosis surveillance system in the provinces of Santa Fe, Entre Ríos and Buenos Aires (year 2016)

INTRODUCCIÓN La leptospirosis es una de las zoonosis de mayor prevalencia mundial. Es sub-diagnosticada debido a las formas clínicas leves, a la falta de conocimiento de la enfermedad, a la falta de accesibilidad y a las desventajas de las técnicas diagnósticas. Las provincias de Santa Fe, Entre Ríos y Buenos Aires concentran el mayor número de casos. OBJETIVOS Ampliar el sistema de vigilancia intensificada a 9 establecimientos de salud pública y privada de las provincias mencionadas. MÉTODOS Se incluyeron 64 pacientes con sospecha de leptospirosis (1 fallecidos), desde enero de 2016 hasta abril de 2017, a los que se les realizó TR, MAT, IgM-Elisa, PCR en tiempo real y cultivo. RESULTADOS Se obtuvieron 64 primeras muestras, 29 segundas y 7 terceras. Se confirmaron 11 casos por MAT (3), PCR en tiempo real (3) y por ambas técnicas (5). La PCR en tiempo real permitió la confirmación precoz, a partir del 2º día de evolución, en 8 pacientes. Las técnicas de screening (TR, IgM-Elisa) fueron positivas en todos los casos. Se aisló una cepa perteneciente al serogrupo Canicola. La tipificación molecular de las muestras positivas por PCR en tiempo real presentó similitud con L. interrogans, a 4 de ellos se les pudo definir el serogrupo; Pyrogenes (2), Icterohaemorrhagiae y Canicola. En los pacientes confirmados la clínica y epidemiologia de la enfermedad coincide con lo descripto por otros autores en nuestro país. DISCUSIÓN Los resultados de este trabajo demuestran la relevancia de la sospecha clínica-epidemiológica de la enfermedad, remarcan la importancia de la obtención de segundas muestras y la utilidad de las técnicas de screening en los laboratorios de la Red Nacional de Leptospirosis. También demuestran la precocidad del diagnóstico utilizando PCR en tiempo Real y la necesidad de implementar el cultivo de muestras clínicas para el aislamiento de Leptospira spp, de vital importancia para la prevención y control de la enfermedad.

Isolation and clinical sample typing of human leptospirosis cases in Argentina.

Leptospira typing is carried out using isolated strains. Because of difficulties in obtaining them, direct identification of infective Leptospira in clinical samples is a high priority. Multilocus sequence typing (MLST) proved highly discriminatory for seven pathogenic species of Leptospira, allowing isolate characterization and robust assignment to species, in addition to phylogenetic evidence for the relatedness between species. In this study we characterized Leptospira strains circulating in Argentina, using typing methods applied to human clinical samples and isolates. Phylogenetic studies based on 16S ribosomal RNA gene sequences enabled typing of 8 isolates (6 Leptospira interrogans, one Leptospira wolffii and one Leptospira broomii) and 58 out of 85 (68.2%) clinical samples (55 L. interrogans, 2 Leptospira meyeri, and one Leptospira kirschneri). MLST results for the L. interrogans isolates indicated that five were probably Canicola serogroup (ST37) and one was probably Icterohaemorrhagiae serogroup (ST17). Eleven clinical samples (21.6%), provided MLST interpretable data: five were probably Pyrogenes serogroup (ST13), four Sejroe (ST20), one Autumnalis (ST22) and one Canicola (ST37). To the best of our knowledge this study is the first report of the use of an MLST typing scheme with seven loci to identify Leptospira directly from clinical samples in Argentina. The use of clinical samples presents the advantage of the possibility of knowing the infecting strain without resorting to isolates. This study also allowed, for the first time, the characterization of isolates of intermediate pathogenicity species (L. wolffii and L. broomii) from symptomatic patients.

[Evaluation of a reduced panel of leptospira strains for microagglutination]. / Evaluación de un panel reducido de cepas de leptospiras para el diagnóstico en humanos por microaglutinación (MAT).

OBJECTIVE: To evaluate if the use of the 19 Leptospira strains panel suggested by the International Leptospirosis Society of World Health Organization for microagglutination allows confirmation of more cases that the 12 strains panel used in Argentina. MATERIALS AND METHODS: Cross-sectional observational study. We studied 441 serum samples corresponding to Argentinean patients with suspected leptospirosis derived during from July to December, 2009 and from January to October, 2013. RESULTS: The same number of positive samples was obtained using the MAT with the 19 or 12 strains. In six cases a serovar of the expanded collection was presumably infecting, but always coagglutinated with strains of the reduced panel. CONCLUSION: In Argentina, the diagnosis of leptospirosis by MAT could be made using the reduced 12 strains panel, obtaining the same result in case detection as using the 19 strains panel. Additional information provided by the use of all strains could be the presumably infecting serogroup.

Evaluación de un panel reducido de cepas de leptospiras para el diagnóstico en humanos por microaglutinación (MAT) / Evaluation of a reduced panel of leptospira strains for microagglutination

Objetivo. Evaluar si el uso del panel de 19 cepas de leptospiras, sugerido por la Sociedad Internacional de Leptospirosis para la microaglutinación (MAT, por sus siglas en inglés), permite mayor confirmación de casos que el de 12 cepas. Material y métodos. Estudio observacional de corte transversal. Se estudiaron 441 muestras de sueros de pacientes de Argentina, derivadas para el diagnóstico de leptospirosis en los periodos de julio de 2009 a diciembre de 2010 y enero a octubre de 2013. Resultados. Se obtuvo el mismo resultado con el panel reducido que con el ampliado. En seis casos resultó presumiblemente infectante algún serovar del panel ampliado, aunque siempre coaglutinando con cepas del reducido. Conclusión. En Argentina, el diagnóstico de leptospirosis por MAT podría continuar realizándose con el panel reducido, lo que reduciría el costo y tiempo de diagnóstico. La información adicional que aportaría el panel ampliado está relacionada con la epidemiología, mediante un mejor conocimiento del serogrupo presumiblemente infectante.

Objective. To evaluate if the use of the 19 Leptospira strains panel suggested by the International Leptospirosis Society of World Health Organization for microagglutination allows confirmation of more cases that the 12 strains panel used in Argentina. Materials and methods. Cross-sectional observational study. We studied 441 serum samples corresponding to Argentinean patients with suspected leptospirosis derived during from July to December, 2009 and from January to October, 2013. Results. The same number of positive samples was obtained using the MAT with the 19 or 12 strains. In six cases a serovar of the expanded collection was presumably infecting, but always coagglutinated with strains of the reduced panel. Conclusion. In Argentina, the diagnosis of leptospirosis by MAT could be made using the reduced 12 strains panel, obtaining the same result in case detection as using the 19 strains panel. Additional information provided by the use of all strains could be the presumably infecting serogroup.

Diseño y aplicación de una nueva herramienta para la evaluación de la Red Nacional de Laboratorios de Leptospirosis / Design and application of a new tool for the evaluation of the national betwork of leptospirosis laboratories

INTRODUCCION: La leptospirosis es unaenfermedad febril aguda, frecuentemente subdiagnosticada ysubnotificada por su diversa presentación clínica, baja sospechay falta de disponibilidad de métodos diagnósticos rápidos, simples y eficientes. El Laboratorio Nacional de Referencia de Leptospirosis (LNRL) del Instituto Nacional de EnfermedadesRespiratorias (INER) ôDr. Emilio Coniö produce antígeno termorresistente (TR) y lo distribuye a los laboratorios de la Red Nacional de Laboratorios de Leptospirosis (RNLL). Luego, éstosrealizan la técnica y envían las muestras positivas al LNRL para la confirmación del diagnóstico y las negativas para control de calidad (CC). Sin embargo, hasta la realización de este estudio noexistía un control de uso de TR y envío de muestras. OBJETIVOS: Diseñar y aplicar una herramienta para evaluar operativamente la RNLL. METODOS: Durante 2012 y 2013 se aplicó una planilla de registro de uso de TR, y se crearon y analizaron indicadores de eficiencia de uso, aplicación y reposición del TR y derivación de muestras. RESULTADOS: Se observó que entre 2012 y 2013 mejoró la participación en el CC y la eficiencia de uso del TR.La derivación de muestras positivas y la solicitud de reposición de TR se mantuvieron similares. CONCLUSIONES: La planilladiseñada permitió obtener información de cantidad y calidad de uso del TR, y calcular indicadores para evaluar su distribución y aplicación. Se observó que el desempeño de los laboratoriosmejoró de un año al otro.

INTRODUCTION: Leptospirosis is an acute febrile disease, often underdiagnosed and underreported because of its diverse clinical presentation, low suspicion, and unavailability of rapid, simple and efficient diagnostic methods. TheNational Leptospirosis Reference Laboratory (LNRL) of the National Institute of Respiratory Diseases ôDr. Emilio Coniö produces thermo-resistant antigen (TR) and distributes it to the National Network of Leptospirosis Laboratories (RNLL). These laboratories perform the screening, send the positivesamples for confirmation and the negative ones for quality control. However, until this study there was no control of TR uses and shipment of samples. OBJECTIVES: To design andimplement a tool to operatively assess the RNLL. METHODS: During the years 2012 and 2013, a registration form for the TR use was applied, also creating and analyzing indicators of use efficiency, application and replacement of TR, and referral of samples. RESULTS: It was noted that in the period2012-2013, the participation in the CC and efficiency of use of TR improved. The derivation of positive samples and request for replacement TR remained similar. CONCLUSIONS: The designed form allowed for the first time to obtain information on the quantity and quality of use of the reagent, to design andcalculate indicators to assess the distribution and application of TR during 2012 and 2013. In general, the performance of decentralized laboratories improved from one year to the next.

Vigilancia intensificada de Leptospirosis en Santa Fe y Entre Ríos (2012-2013) / Leptospirosis Enhanced Surveillance System in Santa Fe and Entre Rios (2012-2013)

La leptospirosis es la zoonosis de mayor prevalencia mundial. Santa Fe y Entre Ríos concentran la mayoría de casos en Argentina. OBJETIVO: Implementar y describir un sistema de vigilancia intensificada de casos de leptospirosis en Santa Fe y Entre Ríos. MÉTODOS: La investigación, desarrollada desde enero de 2012 hasta marzo de 2013, implicó la intervención en sistemas y servicios de salud, así como el fortalecimiento de la red de laboratorios para el diagnóstico específico y aislamiento de leptospiras. La información obtenida a partir de las fichas clínico-epidemiológicas de los casos confirmados fue analizada y presentada mediante medidas de resumen. La vigilancia intensificada se implementó en siete hospitales estratégicos. RESULTADOS: Ingresaron 183 pacientes. Se confirmaron 24 casos (13%): 10 por MAT, 9 por PCR en tiempo real y 5 por ambos métodos. Se obtuvieron 3 aislamientos (sero grupo Canicola). Fallecieron 4 pacientes con hemorragia pulmonar sumada a compromiso renal, hepático y/o plaquetopenia. CONCLUSIONES: La vigilancia intensificada permitió obtener aislamientos humanos, ratificó el valor de la MAT, evidenció la utilidad de PCR en tiempo real para diagnóstico precoz y corroboró la dificultad de obtener segundas muestras. La presentación clínica evidenció una elevada mortalidad global y una alta frecuencia de compromiso respiratorio asociado a disfunciones orgánicas múltiples de aparición precoz...

Leptospirosis is the most prevalent zoonosis worldwide. In Argentina, Santa Fe and Entre Ríos are the provinces where most cases occur. OBJECTIVE:To implement and describe a system of enhanced surveillance of leptospirosis in Santa Fe and Entre Ríos. METHODS: The enhanced surveillance was carried out from January 2012 to March 2013. The study involved the intervention in health services and systems as well as the laboratory network strengthening for specific diagnosis and leptospira isolation. The information collected from clinical epidemiological records of confirmed cases was analyzed and presented through summary measures. The surveillance was implemented in seven strategic hospitals. RESULTS: A total of 183 patients were enrolled, and 24 cases (13%)were confirmed: 10 through MAT, 9 through real time PCR and 5 through both methods. It was possible to obtain 3 isolates (Canicola serogroup). 4 patients died with pulmonary hemorrhage coupled with renal impairment, hepaticand/ or thrombocytopenia. CONCLUSIONS: The enhanced surveillance allowed to obtain human isolates, confirmed the diagnostic value of MAT, showed the utility of real time PCR for early diagnosis and the difficulties to obtain second samples. The clinical presentation revealed a high global mortality and a high frequency of respiratory involvement related to multiple organ dysfunctions from early stages...