Enzyme immunoassay using Leishmania (Viannia) braziliensis antigens for laboratorial diagnosis of American cutaneous leishmaniasis

Vidigal, Camila de Pádua; Marcussi, Viviane Mathias; Marcussi, Lílian Mathias; Mikcha, Jane Martha Graton; Arraes, Sandra Mara Alessi Aristides; Lonardoni, Maria Valdrinez Campana; Silveira, Thaís Gomes Verzignassi

doi:10.1016/j.actatropica.2008.04.026

Cited by 11 publications

(3 citation statements)

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“…Cross-reactivity with other diseases, such as Chagas disease, toxoplasmosis and visceral leishmaniasis is one of the most frequent problems in studies of diagnostic tests for CL [ 20 , 28 – 30 ]. One of the reasons for high rates of cross-reactivity in serological tests is due to the fact that many organisms share proteins with high degrees of homology.…”

Section: Discussionmentioning

confidence: 99%

“…braziliensis promastigotes was used, investigators found a sensitivity of 85.41% and a specificity of 92.42% in comparison to other diseases, including Chagas disease, toxoplasmosis and paracoccidioidomycosis. However, this test had a high rate of false positive results among healthy controls (10.4%) [ 30 ]. Flow cytometry for the diagnosis of leishmaniasis has been compared with other tests in addition to the ELISA, such as an indirect immunofluorescence test.…”

Section: Discussionmentioning

confidence: 99%

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Detection of IgG Anti-Leishmania Antigen by Flow Cytometry as a Diagnostic Test for Cutaneous Leishmaniasis

et al. 2016

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Diagnosis of cutaneous leishmaniasis (CL) relies on clinical presentation, parasite isolation, histopathologic evaluation and positive Montenegro skin test. However, the low amounts of parasites in the lesion of these individuals make parasite isolation and histopatologic diagnosis unreliable, often leading to false-negative results. Also, 15% of people living in endemic areas have sub-clinical infection characterized by positive Montenegro skin test, which may contribute to misdiagnosis. Although the main Leishmania killing mechanism is through cell-mediated immune response, antibodies against Leishmania antigens are found in infected individuals. Here our goal was to develop a new serological technique using polystyrene microspheres sensitized with soluble Leishmania antigens as a tool for the detection of IgG in serum from CL patients by flow cytometry. To validate the assay we carried out a comparative test (ELISA) commonly used as a diagnostic test for parasitic diseases. To determine cross-reactivity we used serum from patients with Chagas disease, caused by a trypanosome that has several proteins with high homology to those of the Leishmania genus. We observed that the flow cytometry technique was more sensitive than the ELISA, but, less specific. Our results show that the flow cytometry serologic test can be used to confirm CL cases in L. braziliensis transmission areas, however, presence of Chagas disease has to be ruled out in these individuals.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Detection of IgG Anti-Leishmania Antigen by Flow Cytometry as a Diagnostic Test for Cutaneous Leishmaniasis

et al. 2016

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“…In this way, a polymorphic specific-PCR (PS-PCR) approach developed and directly applied on clinical samples, and the sequencing of cytb gene have shown promising results [5,10]. However, these techniques are difficult to apply in rural health centers of endemic areas, because of the technical requirements and the relatively high cost, which still limit its routine use [11].…”

Section: Introductionmentioning

confidence: 99%

High performance of an enzyme linked immunosorbent assay for American tegumentary leishmaniasis diagnosis with Leishmania (Viannia) braziliensis amastigotes membrane crude antigens

et al. 2020

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The diagnosis of American tegumentary leishmaniasis (ATL) still requires the design of more effective tools. Leishmania (Viannia) braziliensis is the causal agent of the 90% of Argentinean ATL cases. Considering the current knowledge, an ELISA based crude antigen (CA) for the diagnosis was designed. Ninety-nine subjects diagnosed as ATL, 27 as no-ATL, and 84 donors from non-ATL-endemic areas were included in this study. The current ATL diagnosis was based four techniques, dermal smear microscopic examination (parasitological test), PCR, Leishmanin skin test, and clinical records. We obtained CA extracts from promastigotes and amastigotes from macrophage cultures of different zymodemes of endemic Leishmania species circulating in the study area. Crude antigens from the 'local' main zymodeme of L. (V.) braziliensis showed the highest reactivity against anti-Leishmania antibodies compared to the other included species. The CA of amastigotes of this zymodeme was 3.4 fold more reactive than promastigotes one. Moreover, amastigote-membrane CA (MCA) were 3.6 fold more reactive than the soluble antigens. The MCA-ELISA reached a sensitivity and specificity of 98% (CI = 94.7%-100%) and 63.6% (53.9-73.1), respectively. When anti-Trypanosoma cruzi reactive sera were excluded, the specificity reached 98.4% (94.4-100), while the sensitivity was similar, with a positive predictive value (PV) of 98.6% (94.6-100) and negative PV of 96.3% (91.6-100). The performance of the MCA-ELISA results strongly contribute to the final diagnostic decision, since a non-reactive serological result almost discards the suspected ATL, because of its high negative PV. The developed PLOS ONE

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Immunoenzymatic assay for the diagnosis of American tegumentary leishmaniasis using soluble and membrane‐enriched fractions from infectious Leishmania (Viannia) braziliensis

Cataldo¹,

Mello²,

Mouta-Confort³

et al. 2010

Clinical Laboratory Analysis

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The diagnosis of American tegumentary leishmaniasis (ATL) is based on the visualization or isolation of the parasite, which is a time-consuming and poorly sensitive method. In this study, we evaluated the accuracy and reliability of ELISA for the diagnosis of ATL using soluble (SF) and membrane-enriched (MF) antigen fractions obtained from an infectious strain of Leishmania (Viannia) braziliensis. A total of 152 serum samples investigated at a referral center in Rio de Janeiro, Brazil, between 2005 and 2007 were studied. Each sample was tested twice with each fraction for the calculation of reliability (intraclass coefficient (ICC)). Cut-off values of 0.22 (SF) and 0.33 (MF) were defined. The use of the fractions resulted in good discrimination between patients, with a large area under the curve (P<0.0001), but no difference was observed between the two fractions (P=0.45). Sensitivity was 89.5% for each fraction, specificity was 89.5% for SF and 93.4% for MF, and the positive likelihood ratio was 8.5 for SF and 13.6 for MF. The ICCs were excellent (SF: 0.96 and MF: 0.90). The antigens tested provided precision and accuracy for the diagnosis of ATL, with SF being recommended due to its lower cost and greater practicality.

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Enzyme immunoassay using Leishmania (Viannia) braziliensis antigens for laboratorial diagnosis of American cutaneous leishmaniasis

Cited by 11 publications

References 16 publications

Detection of IgG Anti-Leishmania Antigen by Flow Cytometry as a Diagnostic Test for Cutaneous Leishmaniasis

Detection of IgG Anti-Leishmania Antigen by Flow Cytometry as a Diagnostic Test for Cutaneous Leishmaniasis

High performance of an enzyme linked immunosorbent assay for American tegumentary leishmaniasis diagnosis with Leishmania (Viannia) braziliensis amastigotes membrane crude antigens

Immunoenzymatic assay for the diagnosis of American tegumentary leishmaniasis using soluble and membrane‐enriched fractions from infectious Leishmania (Viannia) braziliensis

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