Comparison of flow cytometry and indirect immunofluorescence assay in the diagnosis and cure criterion after therapy of American tegumentary leishmaniasis by anti-live Leishmania (Viannia) braziliensis immunoglobulin G

Oliveira, Andresa Pereira de; Castro, Maria Carolina Accioly Brelaz de; Almeida, André; Souza, Marina de Assis; Oliveira, Beatriz Coutinho de; Reis, Luiza Campos; Goto, Hiro; Brito, Maria Edileuza Felinto de; Celeste, Beatriz Julieta; Martins‐Filho, Olindo Assis; Pereira, Valéria Rêgo Alves

doi:10.1016/j.jim.2012.11.002

Cited by 12 publications

(5 citation statements)

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“…Thus, the low reactivity of antibodies showed by CUR patients in the present work suggests that our method can be useful for clinical cure monitoring. These findings agree with other authors that have also shown that low levels of antibodies after therapy can indicate a successful cure [37–39].…”

Section: Discussionsupporting

confidence: 93%

American tegumentary leishmaniasis diagnosis using L. (V.) braziliensis fixed promastigotes: a comparative performance of serological tests and spontaneous cure identification

et al. 2019

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BackgroundThe present study aimed to demonstrate the applicability of a flow cytometry-based serology approach to identify spontaneous cure by the detection of immunoglobulin G, and also, the diagnosis and cure criterion by the IgG1 isotype in American Tegumentary Leishmaniasis – ATL caused by L. (V.) braziliensis. Also, a comparison between flow cytometry with the serological conventional technique was performed.MethodsForty five individuals were included in study. They were assessed in two moments: First, 8 subjects spontaneously cured of ATL, 8 healthy individuals and 15 patients who had a positive diagnosis for ATL were selected before treatment to identify spontaneous cure by immunoglobulin G detection. Secondly, 14 patients who were positive for ATL were selected and had their blood collected before and 1, 2 and 5 years after treatment, respectively, for the diagnostic tests (ELISA and flow cytometry) and cure criterion evaluation using the IgG1 isotype.ResultsThe analysis of the mean percentage of positive fluorescent parasites (PPFP) along with the titration curves of IgG anti-fixed promastigotes of L.(V.)braziliensis, confirmed the applicability of this method for monitoring spontaneous cure in ATL with outstanding co-positivity (100%) and co-negativity (100%) performance indexes. Regarding the results of the comparison between flow cytometry and ELISA it was seen that there was a better accuracy of the first one in relation to the other. When IgG1 applicability was evaluated, it was observed that before treatment, 36.8% of the patients were negative; in patients 1 year post-treatment, 82.3%; 2 years post-treatment, 27.2% and in patients 5 years post-treatment, 87.5%. The overall analysis of the results suggests that flow cytometry can be applied to ATL detection, and that the use of IgG1 isotype has possibilities to contribute as a more specific diagnostic method.ConclusionsTherefore, this area has great perspectives use for the diagnosis and cure criterion, and also it can be scaled up with the possibility to characterize the different clinical stages of the disease. Together, these findings demonstrate the applicability of a flow cytometry-based serology approach and opens up new avenues of research with this technique, such as the understanding the humoral response in ATL patients.

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Section: Discussionsupporting

confidence: 93%

American tegumentary leishmaniasis diagnosis using L. (V.) braziliensis fixed promastigotes: a comparative performance of serological tests and spontaneous cure identification

et al. 2019

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“…To our knowledge, this is the first study using the detection of antibodies anti-L. infantum by flow cytometry for VL diagnosis in HIV-infected individuals. Previous reports started by using this technique to evaluate IgG binding to live promastigotes to assay individuals with VL (24,25) and with American tegumentary leishmaniasis (26). The used of fixed cells as an alternative was also investigated with both Chagas disease and tegumentary leishmaniasis (27,28).…”

Section: Discussionmentioning

confidence: 99%

A Flow Cytometry-Based Serological Assay to Detect Visceral Leishmaniasis in HIV-Infected Patients

et al. 2021

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Visceral Leishmaniasis (VL) is a severe parasitic disease that has emerged as an important opportunistic condition in HIV-infected patients and whose control is impaired by inaccurate identification. This is mainly due to the serological tests used for VL having a reduced performance in cases of VL-HIV coinfection due to a low humoral response. In this situation, however, a positive test has even greater diagnostic value when combined with the clinical status. This study aimed to evaluate the application and performance of flow cytometry to detect anti-Leishmania infantum antibodies in HIV-infected patients. Sera from VL/HIV coinfected patients, characterized using “gold standard” techniques, were compared with sera from healthy controls plus sera from HIV-infected individuals. The flow cytometry results were expressed as levels of IgG reactivity, based on the percentage of positive fluorescent parasites (PPFP). A ROC curve analysis of a serum titration indicated a PPFP of 1.26% as being the cutoff point to segregate positive and negative results. At the 1:2,048 dilution, with 89% sensitivity and 83% specificity, flow cytometry showed greater sensitivity in relation to the serological tests evaluated. Futhermore, flow cytometry was the only assay that positively identified all VL-HIV patients with quantified HIV load. Together, these findings suggest that flow cytometry may be used as an alternative serological approach for VL identification and as a tool to characterize the humoral response against Leishmania infantum in HIV-infected patients.

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“…Flow cytometry for the diagnosis of leishmaniasis has been compared with other tests in addition to the ELISA, such as an indirect immunofluorescence test. These studies also showed that the flow cytometry method had higher sensitivity and lower specificity in relation to indirect immunofluorescence [ 22 , 32 ].…”

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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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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Comparison of flow cytometry and indirect immunofluorescence assay in the diagnosis and cure criterion after therapy of American tegumentary leishmaniasis by anti-live Leishmania (Viannia) braziliensis immunoglobulin G

Cited by 12 publications

References 33 publications

American tegumentary leishmaniasis diagnosis using L. (V.) braziliensis fixed promastigotes: a comparative performance of serological tests and spontaneous cure identification

American tegumentary leishmaniasis diagnosis using L. (V.) braziliensis fixed promastigotes: a comparative performance of serological tests and spontaneous cure identification

A Flow Cytometry-Based Serological Assay to Detect Visceral Leishmaniasis in HIV-Infected Patients

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

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