Duplex qPCR for Leishmania species identification using lesion imprint on filter paper.

BACKGROUND: American cutaneous leishmaniasis (ACL) is caused by different Leishmania parasites, which stimulate and direct the immune response against the infection. OBJECTIVE: To evaluate the TaqMan probe technology applicability to diagnose and identifying of Leishmania spp. related to the ACL etiology. METHODOLOGY: Through the MEGA 6.0 software, performed an in silico analysis using multiple alignments of Leishmania spp. which were available on GenBank for different genomic targets. The efficiency (e), specificity and detection limit (DL) were calculated for each system, these were associated to compose a duplex-qPCR (DqPCR). The samples of blood, lesion biopsy and lesion imprint on filter paper from patients residing in states of Amazonas (AM) and Pernambuco (PE)-Brazil, (cases and controls) were used to perform the DqPCR technique. The capacity to identify the Leishmania species was determined by comparison with isoenzymes method and sequencing analysis. RESULTS: Internal Transcribed Spacer 1 (rDNA) was the target selected. Two sets of primers and probes were designed and combined: SVS for subgenus Viannia and LaS for L. (L.) amazonensis. The results were: SVSe = 93.24%, SVS DL = 50 fg/µL; LaSe = 89.3%, LaSLD = 5 fg/µL presented 100% of specificity. In total, 236 individuals participated of the present study, wherein were 101 blood samples, 33 biopsies and 147 lesion imprints. The imprint was the most sensitive sample, showing 83.06% of sensitivity, 86.96% of specificity and substantial agreement between the techniques analysis (k = 0.531; p < 0,001). Regarding the species identification, DqPCR and sequencing/isoenzymes have agreed at 100%, since the infection is caused by a single Leishmania species. CONCLUSION: The DqPCR technique was applicable in diagnosis and identification of Leishmania spp. (subgenus Viannia and L. amazonensis). Furthermore, the lesion imprint is less invasive, allowing a fewer discomfort and greater acceptance by the patients, in addition of being low cost and easy handling.

Clinical and epidemiological profiles of patients with American cutaneous leishmaniasis from the states of Pernambuco and Amazonas, Brazil.

INTRODUCTION: Brazil has a high number of cases of American cutaneous leishmaniasis (ACL) in the north and northeast regions. Therefore, continuous surveillance of environmental and socioeconomic factors in endemic areas is needed to develop strategic control measures. This study aimed to describe the clinical and epidemiological profiles of patients with ACL. METHODS: All patients were from the states of Amazonas and Pernambuco, and examinations were carried out between 2015 and 2018. All patients had a clinical and epidemiological history compatible with ACL after positive diagnostic tests. Information obtained from medical records included gender, employment activity, level of education, age, and number and sites of lesions. RESULTS: A total of 213 patients were included, of whom 30.98% were female and 69.02% were male. The main employment activity was agriculture (27.56%). The most common level of education was elementary (62.42%). The average age was approximately 39 years. The majority of the patients presented only with one lesion (54.87%), and legs/feet were the most commonly affected area (48.25%), followed by the arms/hands (44.75%). CONCLUSIONS: These data demonstrated that irrespective of the patients' places of origin, interventions need to be focused on men of economically productive age, in view of the high risk of exposure to the vector in this group. Education activities need to be directed to farmers about the importance of protection against ACL vectors during work. Such information must also be directed to employers as a way of implementing and maintaining appropriate working conditions and stepping up vector control.

Report of human fascioliasis under different climatic conditions in Brazil / Registro de fasciolose humana em diferentes condições climáticas no Brasil / Registro de fascioliasis humana bajo diferentes condiciones climáticas en Brasil

Introduction: Human fascioliasis is a plant-borne and water-borne infection caused by the trematodes Fasciola hepatica and Fasciola gigantica. It is one of the main neglected tropical diseases, and infections in humans occur via the ingestion of contaminated water and food. This study reviews all the recorded cases of human fascioliasis in Brazil under different climatic conditions in the national territory. Methodology:A survey of human fascioliasis cases in Brazil was carried out using the Google Scholar, Lilacs and PubMed databases. The climatic variables such as temperature, precipitation, moisture and altitude were obtained from the database of the Instituto Nacional de Meteorologia (INMET). Results: Between the years 1958 and 2022, sixty-six cases of human fascioliasis were recorded in places with temperature levels between 22 °C to 33 °C, humidity 78% to 86%, precipitation 90 mm to 167 mm, and at an altitude of 16 to 935 meters above sea level. Conclusion: The parasite's ability to adapt to different climatic conditions is observed in Brazil and the number of cases of human fascioliasis in the national territory may be higher due to underreporting related to the difficulty in diagnosing the infection.

Introdução: A fasciolíase humana é uma infecção de origem vegetal e hídrica, causada pelos trematódeos Fasciola hepatica e Fasciola gigantica. É uma das principais doenças tropicais negligenciadas, e as infecções em humanos ocorrem através da ingestão de água e alimentos contaminados. Este estudo revisa todos os casos registrados de fasciolíase humana no Brasil sob diferentes condições climáticas no território nacional. Metodologia: Um levantamento dos casos de fasciolíase humana no Brasil foi realizado nas bases de dados Google Scholar, Lilacs e PubMed. As variáveis climáticas como temperatura, precipitação, umidade e altitude foram obtidas do banco de dados do Instituto Nacional de Meteorologia (INMET). Resultados: Entre os anos de 1958 e 2022, sessenta e seis casos de fasciolíase humana foram registrados em locais com níveis de temperatura entre 22 °C a 33 °C, umidade de 78% a 86%, precipitação de 90 mm a 167 mm e altitude de 16 a 935 metros acima do nível do mar. Conclusão: A capacidade de adaptação do parasito a diferentes condições climáticas é observada no Brasil e o número de casos de fasciolíase humana no território nacional pode ser maior devido à subnotificação relacionada à dificuldade de diagnóstico da infecção.

Introducción: La fascioliasis humana es una infección de origen vegetal y acuático, causada por los trematodos Fasciola hepatica y Fasciola gigantica. Es una de las principales enfermedades tropicales desatendidas, y las infecciones en humanos ocurren a través de la ingestión de agua y alimentos contaminados. Este estudio revisa todos los casos registrados de fascioliasis humana en Brasil bajo diferentes condiciones climáticas en el territorio nacional. Metodología: Se realizó una encuesta de casos de fascioliasis humana en Brasil utilizando las bases de datos Google Scholar, Lilacs y PubMed. Las variables climáticas como temperatura, precipitación, humedad y altitud se obtuvieron de la base de datos del Instituto Nacional de Meteorología (INMET). Resultados: Entre los años 1958 y 2022 se registraron sesenta y seis casos de fascioliasis humana en lugares con temperatura entre 22 °C a 33 °C, humedad entre 78% y 86%, precipitación entre 90 mm y 167 mm y una altitud de 16 a 935 metros sobre el nivel del mar. Conclusión: La capacidad de adaptación del parásito a diferentes condiciones climáticas se observa en Brasil y el número de casos de fascioliasis humana en el territorio nacional puede ser mayor debido al subregistro relacionado con la dificultad en el diagnóstico de la infección.

Clinical and epidemiological profiles of patients with American cutaneous leishmaniasis from the states of Pernambuco and Amazonas, Brazil

Abstract INTRODUCTION: Brazil has a high number of cases of American cutaneous leishmaniasis (ACL) in the north and northeast regions. Therefore, continuous surveillance of environmental and socioeconomic factors in endemic areas is needed to develop strategic control measures. This study aimed to describe the clinical and epidemiological profiles of patients with ACL. METHODS: All patients were from the states of Amazonas and Pernambuco, and examinations were carried out between 2015 and 2018. All patients had a clinical and epidemiological history compatible with ACL after positive diagnostic tests. Information obtained from medical records included gender, employment activity, level of education, age, and number and sites of lesions. RESULTS: A total of 213 patients were included, of whom 30.98% were female and 69.02% were male. The main employment activity was agriculture (27.56%). The most common level of education was elementary (62.42%). The average age was approximately 39 years. The majority of the patients presented only with one lesion (54.87%), and legs/feet were the most commonly affected area (48.25%), followed by the arms/hands (44.75%). CONCLUSIONS: These data demonstrated that irrespective of the patients' places of origin, interventions need to be focused on men of economically productive age, in view of the high risk of exposure to the vector in this group. Education activities need to be directed to farmers about the importance of protection against ACL vectors during work. Such information must also be directed to employers as a way of implementing and maintaining appropriate working conditions and stepping up vector control.