Diagnosis of American cutaneous leishmaniasis by enzyme immunoassay in patients from Northern Paraná State, Brazil

Abstract

American cutaneous leishmaniasis (ACL) is an endemic disease caused by Leishmania parasites. The ACL diagnosis is commonly accomplished by parasitological and immunological methods. The objective of this work was to evaluate the enzyme immunoassay (EIA) for ACL laboratorial diagnosis. IgG antibodies against Leishmania (Viannia) braziliensis promastigotes were researched. For the method standardization 240 sera were used: 72 from patients with positive parasitological diagnosis, 38 from normal individuals and 113 from individuals with other pathologies. The sensibility was 93% and the specificity was 70%. Regarding diagnosis, EIA showed 97% positivity in patients with ACL and 97% negativity in patients with other cutaneous lesions. The EIA presented a good performance when used for diagnosis, thus, it may become an important tool for it and for further ACL epidemiological studies in endemic areas.

Investigação sorológica de casos subclínicos de leishmaniose tegumentar após um surto em uma localidade endêmica

“…Os testes sorológicos são pouco utilizados para o diagnóstico da LTA 20 , sendo necessária a associação de diferentes métodos para aumentar a positividade do diagnóstico da doença 16 . Têm Têm Têm sido encontradas algumas dificuldades na utilização do teste intradérmico de Montenegro para o diagnóstico da leishmaniose, como por exemplo, o uso de diferentes espécies do parasita e de uso de diferentes espécies do parasita e de diferentes formas de preparação de antígenos além da capacidade do teste em induzir reações de hipersensibilidade tardia quando realizada pela segunda vez.…”

Serological and Molecular Investigation of Cutaneous Leishmaniasis in Healthy Individuals from an American Cutaneous Leishmaniasis-Endemic Region

“…Parasites were washed in phosphate buffered saline (PBS; pH 7.2), centrifuged at 4°C and 1,700 × g for 10 min, lyophilized and stored at 4°C. The antigens were prepared according to the protocol described by Yoneyama et al (2007). Briefly, the plates were sensitized with a dilution of the extract in carbonate-bicarbonate buffer (0.1 M, pH 9.6); the serum samples were diluted to a rate of 1:150 and added to this plate.…”

“…All samples were analyzed in duplicate. Samples with an average absorbance greater than 0.68 were considered to be positive (Yoneyama et al, 2007). The assay reactivity was confirmed by against the positive and negative controls provided in each plate.…”

“…Enzyme-Linked Immunosorbent Assay (ELISA)-based methods of diagnosis are known to be highly sensitive (Yoneyama et al, 2007) and have been employed in the diagnosis of both clinical and subclinical cases of ACL (Arraes et al, 2008;Szargiki et al, 2009). Previous reports have also confirmed the applicability of Polymerase Chain Reaction (PCR) in the diagnosis of ACL, although it is not routinely used for this purpose (Yoneyama et al, 2007).…”

Presence of anti-Leishmania (Viannia) braziliensis antibodies in blood donors in the West-Central region of the State of Paraná, Brazil

“…Enzyme immunoassay was performed as described by Yoneyama et al (10) using antigens from promastigotes of L. braziliensis (MHOM/BR/1987/M11272). Briefl y, plates were sensitized with the extract crude antigen (10µg/well) diluted in 0.1M carbonate-bicarbonate buffer (pH 9.6).…”