Evidence map of diagnosis, treatment, prognosis, prevention, and control in visceral leishmaniasis / Mapa de evidencia sobre el diagnóstico, tratamiento, pronóstico, prevención y control de la leishmaniasis visceral / Mapa de evidências de diagnóstico, tratamento, prognóstico, prevenção e controle da leishmaniose visceral

ABSTRACT Objective. To develop an evidence map on visceral leishmaniasis prevention, control, diagnosis, treatment, and prognosis. Methods. Systematic reviews on visceral leishmaniasis were searched using MEDLINE/PubMed and Virtual Health Library. After selection, each included systematic review was assessed, characterized, and categorized by intervention type and by outcomes, according to the methodology offered by the PAHO/WHO Latin American and Caribbean Center on Health Sciences Information (BIREME). The methodological quality was assessed using the AMSTAR2 tool to determine the confidence level of the evidence obtained. Results. Among the prevention and control interventions, insecticide spraying, bednets, dog collars, and dog culling were the most assessed, emphasizing that insecticidal dog collars can reduce visceral leishmaniasis incidence in dogs. Regarding diagnosis, polymerase chain reaction (PCR), rK39 immunochromatographic test (rK39 ICT), and direct agglutination test (DAT) presented high sensitivity and specificity. As for treatment, pentavalent antimonials and amphotericin B were the most analyzed drugs and showed therapeutic success; however, serious adverse events can occur due to their use. The prognostic factors identified were anemia, edema, bleeding, jaundice, age, and HIV coinfection. Conclusions. The evidence map developed shows rK39 ICT and DAT as promising diagnostic alternatives and reinforces the efficacy of liposomal amphotericin B and pentavalent antimonials. Insecticide-impregnated dog collars appear as a promising measure for the control of visceral leishmaniasis, but there is also a need for future studies and reviews with higher methodological quality, especially on prevention and control interventions.

RESUMEN Objetivo. Elaborar un mapa de evidencia sobre la prevención, el control, el diagnóstico, el tratamiento y el pronóstico de la leishmaniasis visceral. Métodos. Se realizaron búsquedas de revisiones sistemáticas sobre la leishmaniasis visceral en MEDLINE/PubMed y la Biblioteca Virtual en Salud. Tras la selección, cada revisión sistemática incluida fue sometida a evaluación, caracterización y categorización según tipo de intervención y resultados, de acuerdo con la metodología ofrecida por el Centro Latinoamericano y del Caribe de Información en Ciencias de la Salud de la OPS/OMS (BIREME). La calidad metodológica se evaluó con la herramienta AMSTAR2 para determinar el nivel de confianza de la información obtenida. Resultados. Entre las intervenciones de prevención y control, las más evaluadas fueron la fumigación con insecticidas, los mosquiteros, los collares para perros y el sacrificio de perros y se hizo hincapié en que los collares insecticidas para perros pueden reducir la incidencia de leishmaniasis visceral en perros. En cuanto al diagnóstico, la reacción en cadena de la polimerasa (PCR, por su sigla en inglés), la prueba inmunocromatográfica rK39 (rK39 ICT) y la prueba de aglutinación directa (DAT, por su sigla en inglés) mostraron alta sensibilidad y especificidad. Con respecto al tratamiento, los fármacos más analizados que arrojaron éxito terapéutico fueron los antimoniales pentavalentes y la anfotericina B; sin embargo, su uso puede provocar efectos adversos graves. Los factores pronósticos que se identificaron fueron anemia, edema, sangrado, ictericia, edad y coinfección por el VIH. Conclusiones. El mapa de evidencia elaborado presenta la prueba inmunocromatográfica rK39 y la prueba de aglutinación directa como alternativas diagnósticas prometedoras, y consolida la eficacia de la anfotericina B liposomal y los antimoniales pentavalentes. Los collares de perro impregnados de insecticida parecen ser una medida prometedora para el control de la leishmaniasis visceral, si bien también son necesarios estudios y revisiones adicionales de mayor calidad metodológica, especialmente sobre intervenciones de prevención y control.

RESUMO Objetivo. Desenvolver um mapa de evidências de prevenção, controle, diagnóstico, tratamento e prognóstico da leishmaniose visceral. Métodos. Foram realizadas buscas por revisões sistemáticas sobre leishmaniose visceral no MEDLINE/PubMed e na Biblioteca Virtual em Saúde. Após a seleção, cada revisão sistemática incluída foi avaliada, caracterizada e classificada por tipo de intervenção e por desfechos, de acordo com a metodologia oferecida pelo Centro Latino-Americano e do Caribe de Informação em Ciências da Saúde (BIREME) da OPAS/OMS. A qualidade metodológica foi avaliada utilizando a ferramenta AMSTAR2 para determinar o nível de confiança das evidências obtidas. Resultados. Entre as intervenções de prevenção e controle, pulverização com inseticida, mosquiteiros, coleiras para cães e abate de cães foram as mais frequentemente avaliadas, com destaque para as coleiras inseticidas na redução da incidência de leishmaniose visceral em cães. Quanto ao diagnóstico, a reação em cadeia da polimerase (PCR), o teste imunocromatográfico rK39 (rK39 ICT) e o teste de aglutinação direta (DAT, sigla em inglês para direct agglutination test) apresentaram alta sensibilidade e especificidade. Em relação ao tratamento, os antimoniais pentavalentes e a anfotericina B foram os medicamentos mais analisados e demonstraram sucesso terapêutico, embora seu uso possa resultar em eventos adversos graves. Os fatores prognósticos identificados foram anemia, edema, sangramento, icterícia, idade e coinfecção com HIV. Conclusões. O mapa de evidências desenvolvido mostra o rK39 ICT e o DAT como alternativas promissoras para o diagnóstico e reforça a eficácia da anfotericina B lipossomal e dos antimoniais pentavalentes. As coleiras impregnadas com inseticida aparecem como medida promissora para o controle da leishmaniose visceral, mas estudos e revisões futuras com mais qualidade metodológica, especialmente sobre intervenções de prevenção e controle, são necessários.

C-reactive protein for the diagnosis and prognosis of visceral leishmaniasis caused by Leishmania infantum

Acute-phase reaction (APR) and its marker C-reactive protein (CRP) are heightened in some infectious diseases. APR may contribute to clinical manifestations of systemic inflammation such as hemorrhages, anemia and edema. These symptoms are shared by visceral leishmaniasis (VL), a disease caused by the protozoa Leishmania infantum and L. donovani. The former is zoonotic, hitting mostly children and the immunosuppressed, with higher mortality. As APR and CRP have not been studied in VL caused by L. infantum, we decided to investigate their role as diagnostic and prognostic tools in Brazil. We measured CRP in 136 patients before the treatment of VL and 128 who survived and returned 30 days later and compared it to the clinical presentation, HIV status, and disease severity. Sensitivity for the disease was 97.8% (95%CI: 93.7 - 99.5) and specificity was 85.9% (95%CI: 78.7 - 91.0) with the cut-off of 10mg/L. There was no association of CRP concentration with demographic, clinical and laboratory data. The correlation between pre-and post-treatment levels existed but was poor. Marginal association with the presence of parasites in the bone marrow and death was noticed. The role of APR in the pathogenesis of VL and disease severity remains to be explored. However, the study reveals the significant role of CRP for VL caused by L. infantum and should be routinely required for the diagnosis and follow-up.

Nutritional status and vitamin A and zinc levels in patients with kala-azar in Piauí, Brazil

Abstract INTRODUCTION: Malnutrition and kala-azar (or visceral leishmaniasis) are significant public health problems in different parts of the world. Immunity and susceptibility to infectious and parasitic diseases are directly linked to the host's nutritional state, but little is known about the interaction between nutrition and kala-azar. This study aimed to evaluate nutritional status with kala-azar and correlate these findings with the clinical and laboratory manifestations of the disease, and zinc and retinol levels. METHODS: This was a cross-sectional study of 139 patients with kala-azar. Nutritional status classification was performed according to international recommendations. Parametric or nonparametric tests were applied whenever indicated in a two-sided test with a 5% significance level. RESULTS: Weight loss and malnutrition were more frequent in adults. Body mass index-for-age, fat area of the arm, and upper arm muscle area were significantly associated with probability of death. The presence of human immunodeficiency virus, hepatomegaly, and splenomegaly was correlated with nutritional assessment. Blood leukocyte and lymphocyte, serum creatine, and vitamin A levels were significantly higher in adult men. Vitamin A levels were highly associated with the level of hemoglobin and C-reactive protein (CRP) in multivariate analysis. All patients had reduced plasma zinc levels, but this finding had no association with the outcome variables. CONCLUSIONS: Malnutrition was correlated with severe disease and was more prevalent in older people with kala-azar. Vitamin A deficiency was associated with hemoglobin and CRP. Zinc levels were reduced in patients with kala-azar.

Cutaneous parasitism in patients with American visceral leishmaniasis in an endemic area

Abstract INTRODUCTION: Visceral leishmaniasis (VL) represents a public health concern in several areas of the world. In the American continent, VL transmission is typically zoonotic, but humans with active VL caused by Leishmania infantum are able to infect sandflies. Thus, individuals with cutaneous parasitic infections may act as reservoirs and allow interhuman transmission. Additionally, the skin may be responsible for reactivation of the disease after therapy. This study's objective was to evaluate cutaneous parasitism in humans with VL in an American endemic area. METHODS: A cross-sectional hospital-based study was conducted in northeast Brazil from October 2016 to April 2017. Biopsies of healthy skin for histopathology and immunohistochemistry were performed prior to treatment in all study patients. RESULTS: Twenty-two patients between the ages of five months to 78 years were included in the study. Seven patients (31.8%) tested positive for HIV. Only one patient had cutaneous parasitism, as confirmed by immunohistochemistry prior to treatment. Parasitism was not detected after treatment. CONCLUSIONS: Cutaneous parasitism in the healthy skin of humans with visceral leishmaniasis, although unusual, may be a source of infection for phlebotomine sandflies.

Proposta de abordagem simplificada para suspeitas de meningites: relato de experiência de serviço de referência no estado do Piauí, 2007-2016 / Propuesta para el enfoque inicial simplificado de las sospechas de meningitis: relato de experiencia de servicio de referencia en el estado de Piauí, Brasil, 2007-2016 / Proposal for a simplified approach to suspected meningitis case diagnosis: experience report of a reference service in the state of Piauí, Brazil, 2007-2016

Objetivo: descrever um protocolo simplificado para abordagem de casos suspeitos de meningite. Métodos: relato de experiência na abordagem diagnóstica das meningites no serviço de referência em doenças tropicais do estado do Piauí, Brasil, no período de 2007-2016; extraíram-se informações do Sistema de Informação de Agravos de Notificação (Sinan) e do livro de registro do laboratório; utilizou-se o teste do qui-quadrado para comparar os indicadores da vigilância epidemiológica baseados na classificação final dos casos de meningite; utilizou-se o teste de concordância Phi para verificar a correlação entre os diagnósticos etiológicos presumidos pelo protocolo e aqueles confirmados laboratorialmente. Resultados: considerando-se 4.096 casos de meningite estudados, houve redução na classificação genérica desses casos, de 72 para 47% (p<0,001); a indicação do perfil de investigação laboratorial mostrou concordância com o diagnóstico final dos casos de meningite (rf=0,66; p<0,001). Conclusão: o diagnóstico etiológico específico das meningites foi alcançado em maior proporção durante a utilização do protocolo.

Objetivo: describir un protocolo para el abordaje de casos sospechosos de meningitis. Métodos: se relata la experiencia del diagnóstico de meningitis en el servicio de referencia para enfermedades tropicales del estado de Piauí, Brasil, entre 2007 y 2016; extrajimos informaciones del Sistema de Información de Notificación (Sinan) y del libro de laboratorio; la prueba de chi-cuadrado verificó la alteración de los indicadores de la vigilancia epidemiológica basados en la clasificación de los casos de meningitis; usamos la prueba de concordancia Phi para correlacionar los diagnósticos etiológicos presumidos por el protocolo y aquellos confirmados laboratorialmente. Resultados: considerando los 4.096 casos de meningitis investigados, se observó reducción en la clasificación genérica de casos del 72 al 47% (p<0,001); la selección del perfil de investigación mostró concordancia con la clasificación final de los casos de meningitis (rf=0,66; p<0,001). Conclusión: el diagnóstico etiológico de las meningitis se alcanzó en mayor proporción durante la utilización del protocolo.

Objective: to describe a simplified protocol to diagnose suspected cases of meningitis. Methods: this is an experience report on the approach to diagnosing meningitis at the Tropical Diseases Reference Service in the state of Piauí, Brazil, between 2007 and 2016; information was extracted from the Notifiable Diseases Information System (SINAN) and the laboratory record book; the chi-square test was used to compare epidemiological surveillance indicators based on final meningitis case classification; the Phi coefficient was used to verify the correlation between presumed diagnosis and laboratory-confirmed diagnosis. Results: considering the 4,096 cases of meningitis investigated, there was a reduction in the generic classification of meningitis cases from 72% to 47% (p<0.001); indicated laboratory investigation profile showed agreement with final meningitis case diagnosis. (rf=0.66; p<0.001). Conclusion: a greater proportion of specific etiologic diagnosis of meningitis was achieved while the protocol was in use.

Serological tests fail to discriminate dogs with visceral leishmaniasis that transmit Leishmania infantum to the vector Lutzomyia longipalpis

Abstract INTRODUCTION The control of reservoirs for Leishmania infantum -induced zoonotic visceral leishmaniasis requires the identification of dogs posing a population risk. Here, we assessed the performance of several assays to identify Lutzomyia longipalpis infectious dogs. METHODS We evaluated 99 dogs that were positive for visceral leishmaniasis based on parasite identification. Serological analyses were performed using an enzyme-linked immunosorbent assay, immunofluorescence antibody tests in 1:40 and 1:80 dilutions, rapid dual path platform tests, immunochromatographic assay with a recombinant rK39 antigen, fast agglutination screening tests, and direct agglutination tests. We also performed PCR to analyze peripheral blood and xenodiagnosis. RESULTS Forty-six dogs infected at least one L. longipalpis specimen. Although the serological test sensitivities were above 85% for detecting L. longipalpis infectious dogs, none showed a satisfactory performance, as both specificity (0.06 to 13%) and the area under the receiver operating characteristic curve (45 to 53%) were low. The PCR results were also weak, with a sensitivity of 30%, specificity of 72%, and an area under the receiver operating characteristic curve of 51%. The infected L. longipalpis proportion was higher among asymptomatic dogs than symptomatic dogs. Among the symptomatic dogs, those with ulceration-free skin diseases were more infectious, with an odds ratio of 9.3 (confidence interval of 1.10 - 428.5). The larger the number of insects fed, the greater the detected infectiousness. CONCLUSIONS Our study supports the imperative to develop novel technologies for identifying the infectious dogs that transmit L. infantum for the benefit of public health.

The performance of serological tests for Leishmania infantum infection screening in dogs depends on the prevalence of the disease

ABSTRACT Dogs are considered the main reservoir of Leishmania infantum. This protozoan causes visceral leishmaniasis (VL), an uncontrolled urban zoonosis in Brazil. Serological tests and polymerase chain reaction (PCR) on peripheral blood were performed to identify infected dogs in scenarios of higher and lower prevalence of the disease (Teresina and Vitória). One-hundred infected and 57 non-infected animals from Teresina and 100 non-infected animals from Vitória were studied. Animal selection was not dependent on previous serology. The sensitivity (Teresina) and specificity (Teresina and Vitória) were as follows: indirect antibody fluorescence (IFAT) cut-off of 1:40 (IFAT 1:40): 96%, 18%, and 76%; IFAT 1:80: 90%, 33%, and 93%; direct agglutination test (DAT): 96%, 33%, and 98%; fast agglutination screening test (FAST): 93%, 68%, and 100%; immunochromatographic assay with a recombinant rK39 antigen (rK39): 88%, 74%, and 98%; enzyme linked immunosorbent assay (ELISA): 91%, 79%, and 98%; rapid dual-path platform test (TR DPP®): 98%, 60%, and 98%; and blood PCR: 29%, 93%, and 97%, respectively. In the high transmission area, none of the tests adequately discriminated L. infantum-infected from non-infected dogs. However, in the high transmission city, the area under the receiver operating characteristic (ROC) curve of FAST, DAT, ICrK39, ELISA and TR DPP® was high.

Predicting death from kala-azar: construction, development, and validation of a score set and accompanying software

Abstract INTRODUCTION Early identification of patients at higher risk of progressing to severe disease and death is crucial for implementing therapeutic and preventive measures; this could reduce the morbidity and mortality from kala-azar. We describe a score set composed of four scales in addition to software for quick assessment of the probability of death from kala-azar at the point of care. METHODS: Data from 883 patients diagnosed between September 2005 and August 2008 were used to derive the score set, and data from 1,031 patients diagnosed between September 2008 and November 2013 were used to validate the models. Stepwise logistic regression analyses were used to derive the optimal multivariate prediction models. Model performance was assessed by its discriminatory accuracy. A computational specialist system (Kala-Cal(r)) was developed to speed up the calculation of the probability of death based on clinical scores. RESULTS: The clinical prediction score showed high discrimination (area under the curve [AUC] 0.90) for distinguishing death from survival for children ≤2 years old. Performance improved after adding laboratory variables (AUC 0.93). The clinical score showed equivalent discrimination (AUC 0.89) for older children and adults, which also improved after including laboratory data (AUC 0.92). The score set also showed a high, although lower, discrimination when applied to the validation cohort. CONCLUSIONS: This score set and Kala-Cal(r) software may help identify individuals with the greatest probability of death. The associated software may speed up the calculation of the probability of death based on clinical scores and assist physicians in decision-making.

Visual evoked potentials show strong positive association with intracranial pressure in patients with cryptococcal meningitis / Potenciais evocados visuais mostram correlação positiva forte com a pressão intracraniana em pacientes com meningite criptocócica

Objective : To verify the relationship between intracranial pressure and flash visual evoked potentials (F-VEP) in patients with cryptococcal meningitis. Method The sample included adults diagnosed with cryptococcal meningitis admitted at a reference hospital for infectious diseases. The patients were subjected to F-VEP tests shortly before lumbar puncture. The Pearson’s linear correlation coefficient was calculated and the linear regression analysis was performed. Results : Eighteen individuals were subjected to a total of 69 lumbar punctures preceded by F-VEP tests. At the first lumbar puncture performed in each patient, N2 latency exhibited a strong positive correlation with intracranial pressure (r = 0.83; CI = 0.60 - 0.94; p < 0.0001). The direction of this relationship was maintained in subsequent punctures. Conclusion : The intracranial pressure measured by spinal tap manometry showed strong positive association with the N2 latency F-VEP in patients with cryptococcal meningitis. .

Objetivo : Verificar a relação entre pressão intracraniana e potencial evocado visual por flash (PEV-F) em pacientes com meningite criptocócica. Método A amostra incluiu pacientes admitidos em um hospital de referência para doenças infecciosas. Realizou-se PEV-F antes de cada punção lombar. Calculou-se o coeficiente de correlação de Pearson e a equação de regressão linear entre as variáveis latência N2 e pressão intracraniana inferida através de raquimanometria. Resultados Dezoito pacientes foram submetidos a um total de 69 punções lombares. A latência N2 mostrou correlação positiva forte com a pressão de abertura verificada na primeira punção lombar a que cada paciente foi submetido (r = 0,83; IC = 0,60 – 0,94; p < 0,0001). A positividade da correlação foi mantida nas aferições subsequentes. Conclusão Houve associação positiva forte entre a latência N2 do PEV-F e pressão intracraniana em pacientes com meningite criptocócica. .

Genes that encodes NAGT, MIF1 and MIF2 are not virulence factors for kala-azar caused by Leishmania infantum

Introduction Kala-azar is a disease resulting from infection by Leishmania donovani and Leishmania infantum. Most patients with the disease exhibit prolonged fever, wasting, anemia and hepatosplenomegaly without complications. However, some patients develop severe disease with hemorrhagic manifestations, bacterial infections, jaundice, and edema dyspnea, among other symptoms, followed by death. Among the parasite molecules that might influence the disease severity are the macrophage migration inhibitory factor-like proteins (MIF1 and MIF2) and N-acetylglucosamine-1-phosphotransferase (NAGT), which act in the first step of protein N-glycosylation. This study aimed to determine whether MIF1, MIF2 and NAGT are virulence factors for severe kala-azar. Methods To determine the parasite genotype in kala-azar patients from Northeastern Brazil, we sequenced the NAGT genes of L. infantum from 68 patients as well as the MIF1 and MIF2 genes from 76 different subjects with diverse clinical manifestations. After polymerase chain reaction (PCR), the fragments were sequenced, followed by polymorphism identification. Results The nucleotide sequencing of the 144 amplicons revealed the absence of genetic variability of the NAGT, MIF1 and MIF2 genes between the isolates. The conservation of these genes suggests that the clinical variability of kala-azar does not depend upon these genes. Additionally, this conservation suggests that these genes may be critical for parasite survival. Conclusions NAGT, MIF1 and MIF2 do not alter the severity of kala-azar. NAGT, MIF1 and MIF2 are highly conserved among different isolates of identical species and exhibit potential for use in phylogenetic inferences or molecular diagnosis. .

Identification of blood meal sources of Lutzomyia longipalpis using polymerase chain reaction-restriction fragment length polymorphism analysis of the cytochrome B gene

An analysis of the dietary content of haematophagous insects can provide important information about the transmission networks of certain zoonoses. The present study evaluated the potential of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mitochondrial cytochrome B (cytb) gene to differentiate between vertebrate species that were identified as possible sources of sandfly meals. The complete cytb gene sequences of 11 vertebrate species available in the National Center for Biotechnology Information database were digested with Aci I, Alu I, Hae III and Rsa I restriction enzymes in silico using Restriction Mapper software. The cytb gene fragment (358 bp) was amplified from tissue samples of vertebrate species and the dietary contents of sandflies and digested with restriction enzymes. Vertebrate species presented a restriction fragment profile that differed from that of other species, with the exception of Canis familiaris and Cerdocyon thous. The 358 bp fragment was identified in 76 sandflies. Of these, 10 were evaluated using the restriction enzymes and the food sources were predicted for four: Homo sapiens (1), Bos taurus (1) and Equus caballus (2). Thus, the PCR-RFLP technique could be a potential method for identifying the food sources of arthropods. However, some points must be clarified regarding the applicability of the method, such as the extent of DNA degradation through intestinal digestion, the potential for multiple sources of blood meals and the need for greater knowledge regarding intraspecific variations in mtDNA.

Congenitally transmitted visceral leishmaniasis: report of two brazilian human cases

Visceral leishmaniasis is a relevant public health problem worldwide. Most of the reported cases in Latin America are from Brazil. Herein we report two human cases of congenitally transmitted visceral leishmaniasis in two patients who developed symptoms during pregnancy. The diagnosis was made by visual examination of Leishmania parasites in bone marrow aspirates of the mothers and by detecting parasite kDNA in bone marrow samples of the newborn children using polymerase chain reaction.

Human immunodeficiency virus/Leishmania infantum in the first foci of urban American visceral leishmaniasis: clinical presentation from 1994 to 2010

INTRODUCTION: Human immunodeficiency virus (HIV) coinfection with Leishmania infantum or Leishmania donovani, the agents of visceral leishmaniasis (or kala-azar), has become a fatal public health problem in the tropics where kala-azar is endemic. METHODS: The clinical presentation of patients with HIV and L. infantum coinfection is described using two unique databases that together produce the largest case series of patients with kala-azar infected with HIV in South America. First, a retrospective study paired the list of all patients with kala-azar from 1994 to 2004 with another of all patients with HIV/AIDS from the reference hospital for both diseases in the City of Teresina, State of Piauí, Brazil. Beginning in 2005 through to 2010 this information was prospectively collected at the moment of hospitalization. RESULTS: During the study, 256 admissions related to 224 patients with HIV/L. infantum coinfection were registered and most of them were males between 20-40 years of age. Most of the 224 patients were males between 20-40 years of age. HIV contraction was principally sexual. The most common symptoms and signs were pallor, fever, asthenia and hepatosplenomegaly. 16.8% of the cohort died. The primary risk factors associated to death were kidney or respiratory failure, somnolence, hemorrhagic manifestations and a syndrome of systemic inflammation. The diagnosis of HIV and kala-azar was made simultaneously in 124 patients. CONCLUSIONS: The urban association between HIV and kala-azar coinfection in South America is worrisome due to difficulty in establishing the diagnosis and higher mortality among the coinfected then those with either disease independently. HIV/L. infantum coinfection exhibits some singular characteristics and due to its higher mortality it requires immediate assistance to patients and greater research on appropriate combination therapy. .

Genetic diversity of Leishmania infantum field populations from Brazil

Leishmania infantum (syn. Leishmania chagasi) is the etiological agent of visceral leishmaniasis (VL) in Brazil. The epidemiology of VL is poorly understood. Therefore, a more detailed molecular characterization at an intraspecific level is certainly needed. Herein, three independent molecular methods, multilocus microsatellite typing (MLMT), random amplification of polymorphic DNA (RAPD) and simple sequence repeats-polymerase chain reaction (SSR-PCR), were used to evaluate the genetic diversity of 53 L. infantum isolates from five different endemic areas in Brazil. Population structures were inferred by distance-based and Bayesian-based approaches. Eighteen very similar genotypes were detected by MLMT, most of them differed in only one locus and no correlation was found between MLMT profiles, geographical origin or the estimated population structure. However, complex profiles composed of 182 bands obtained by both RAPD and SSR-PCR assays gave different results. Unweighted pair group method with arithmetic mean trees built from these data revealed a high degree of homogeneity within isolates of L. infantum. Interestingly, despite this genetic homogeneity, most of the isolates clustered according to their geographical origin.

Evaluation of an enzyme immunoassay for clinical diagnosis of neurocysticercosis in symptomatic patients / Avaliação de um teste imunoenzimático para o diagnóstico clínico de neurocisticercose em pacientes sintomáticos

INTRODUCTION: Neurocysticercosis is an infection of the human central nervous system caused by the metacestode larvae of Taenia solium. Neurocysticercosis is the most common parasitic disease in developing countries. Epilepsy is the most common clinical manifestation. Difficulties in confirming the diagnosis motivated the evaluation of the enzyme-linked immunosorbent assay on cerebral spinal fluid (CSF). METHODS: Twenty-two patients with NCC and 44 control patients were studied. CSF was analyzed using a commercial ELISA kit developed for NCC. Sensitivity and specificity were measured and a multivariate logistic regression was performed. RESULTS: Sensitivity and specificity of ELISA were 31.8 percent and 100 percent, respectively, with accuracy of 77.3 percent. Only the size of the lesions proved to be important for performance of the test. CONCLUSIONS: The results showed that ELISA contributes to the diagnosis of neurocysticercosis if the result is negative or if the patient has a lesion of 2 cm or more.

INTRODUÇÃO: Neurocisticercose é a infecção do sistema nervoso central causada pela larva metacestódea da Taenia solium. Neurocisticercose é a parasitose mais comum nos países em desenvolvimento. Epilepsia é a sua manifestação clínica mais comum. A dificuldade para confirmar o diagnóstico motivou a avaliação do ensaio imunoenzimático ligado à enzima no líquido cérebro-espinhal. MÉTODOS: Vinte e dois pacientes com NCC e 44 pacientes controles foram estudados. Líquido cérebro-espinhal foi analisado por um kit ELISA comercial desenvolvido para NCC. A sensibilidade e especificidade foram medidas e uma análise multivariada de regressão logística foi realizada. RESULTADOS: A sensibilidade e a especificidade de ELISA foram, respectivamente, 31,8 por cento e 100 por cento, com acurácia de 77,3 por cento. Apenas o tamanho das lesões mostrou-se importante para o desempenho do teste. CONCLUSÕES: Este estudo concluiu que ELISA contribui para o diagnóstico de NCC, caso o teste seja negativo ou caso o paciente seja portador de lesão cuja dimensão seja igual ou maior que dois centímetros.

Is severe visceral leishmaniasis a systemic inflammatory response syndrome? A case control study / A leishmaniose visceral grave é uma síndrome da resposta inflamatória sistêmica? Um estudo caso-controle

INTRODUCTION: The objective of the study is to identify the main risk factors for death by New World visceral leishmaniasis and establish a coherent pathogenic substrate of severe disease based on clinical findings. METHODS: Seventy-six deceased inpatients and 320 successfully treated inpatients with VL were studied in a case control study. RESULTS: Bacterial infection and bleeding were mutually exclusive events leading to death. Five risk factors were unique for death by bacterial infection (malnutrition, pulmonary rales, severe anemia, severe absolute neutropenia and higher neutrophil count), while another six were unique for death by bleeding (jaundice, severe relative neutropenia, severe thrombocytopenia, liver injury, kidney failure, higher bone marrow parasite load). Bacterial infection, bleeding, severe anemia, diarrhea, dyspnea, edema, jaundice and bone marrow parasite load were the main syndromes of visceral leishmaniasis among successfully treated patients. CONCLUSIONS: The data support the idea that bacterial infections are due to immune paralysis. Broad organ and system involvement is plausibly due to the high production of proinflammatory cytokines, whose actions fit well with visceral leishmaniasis. The syndromes and causative mediators are typical of a slowly developing systemic inflammatory response syndrome.

INTRODUÇÃO: O objetivo do estudo foi i dentificar os principais fatores de risco para morte na leishmaniose visceral do Novo Mundo e estabelecer um substrato patogênico baseado nos achados clínicos coerente para doença grave. MÉTODOS: Em um estudo caso-controle, foram estudados 76 pacientes internados que faleceram e 320 pacientes internados tratados com sucesso. RESULTADOS: Infecção bacteriana e sangramento foram eventos que levaram à morte, mutuamente exclusivos. Cinco fatores de risco foram únicos para morte por infecção bacteriana (desnutrição, estertores pulmonares, anemia grave, neutropenia absoluta grave e número de leucócitos aumentados), enquanto outros seis foram exclusivos para morte por sangramento (icterícia, neutropenia relativa grave, trombocitopenia grave, lesão hepática, insuficiência renal, maior carga de parasitas na medula óssea). Entre os pacientes tratados com sucesso, as principais síndromes de leishmaniose visceral foram infecções bacterianas, sangramento, anemia grave, diarreia, dispneia, edema, icterícia e carga de parasitas na medula óssea. CONCLUSÕES: Os dados apoiam a ideia de que as infecções bacterianas são secundárias a imunoparalisia. O amplo envolvimento de órgãos e sistemas é de forma plausível devido a elevada produção de citocinas pró-inflamatórias, cujas ações se encaixam com a leishmaniose visceral. As síndromes e os mediadores causais são típicos da síndrome de resposta inflamatória sistêmica, desenrolando-se lentamente.

Validação do teste imunocromatográfico rápido IT-LEISH® para o diagnóstico da leishmaniose visceral humana / Validation of the rapid immunochromatographic test IT-LEISH® for the diagnosis of human visceral leishmaniasis

O teste imunocromatográfico rápido IT-LEISH® (DiaMed IT-LEISH®) foi validado para o diagnóstico da leishmaniose visceral (LV) em quatro áreas endêmicas do Brasil. O desempenho do IT-LEISH® foi comparado ao da reação de imunofluorescência indireta; e ao da reação imunoenzimática, usando-se antígeno solúvel de Leishmania chagasi e recombinante K39 (rK39). O estudo incluiu 332 pacientes com quadro clínico sugestivo de LV: 213 casos de LV confirmados parasitologicamente; e 119 não-casos, com confirmação de outra etiologia. O teste IT-LEISH® apresentou sensibilidade de 93 por cento e especificidade de 97 por cento. As técnicas RIFI (imunofluorescência indireta), ELISA L. chagasi e ELISA rK39 apresentaram sensibilidade de 88 por cento, 92 por cento e 97 por cento e especificidades de 81 por cento, 77 por cento e 84 por cento, respectivamente. Os resultados confirmam a validade do teste IT-LEISH® para o diagnóstico da LV no Brasil...

Colelitíase associada ao uso de ceftriaxona / Cholelithiasis associated with the use of ceftriaxone

Crianca de sete anos recebeu ceftriaxona para o tratamento de meningite, evoluindo com dor em hipocôndrio direito associada a cálculo na vesícula biliar. Após três meses, a ultrassonografia abdominal foi normal. O conhecimento de que a ceftriaxona pode levar ao surgimento de colelitíase pode evitar intervencões cirúrgicas desnecessárias.