Performance evaluation of anti-fixed Leishmania infantum promastigotes immunoglobulin G (IgG) detected by flow cytometry as a diagnostic tool for visceral Leishmaniasis

Silva, Elis D.; Oliveira, Beatriz Coutinho de; Oliveira, Andresa Pereira de; Santos, Wagner José Tenório dos; Diniz, George Tadeu Nunes; Neto, Osvaldo P. de Melo; Costa, Carlos Henrique Nery; Silva, Mauro R.B.; Andrade, Luiz Dias de; Medeiros, Zulma; Pereira, Valéria Rêgo Alves

doi:10.1016/j.jim.2019.02.009

Cited by 3 publications

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“…The used of fixed cells as an alternative was also investigated with both Chagas disease and tegumentary leishmaniasis (27,28). In a previous study, we also directly investigated the use of fixed promastigotes to assess IgG binding by flow cytometry for the diagnosis of individuals with VL, reaching a sensitivity of 92-96% in these individuals (15). In the present study, we found a good, but not ideal, sensitivity, although in relation to the conventional tests used for comparison, the sensitivity of flow cytometry was greater.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, it can also be applied to parasitic diseases, such as Chagas Disease and leishmaniasis (13,14). This technique has several advantages for immunoassays, such as high throughput capacity, possibility of analyte quantification, reduced sample volume, high reproducibility and sensitivity (14,15). More importantly, it allows the development of multiplex studies using recombinant antigens, and it can be used as a monitoring tool for cured patients, allowing a more sensitive detection of anti-Leishmania antibodies (16)(17)(18).…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

et al. 2021

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Recent advances and new strategies in Leishmaniasis diagnosis

Brito

Aguiar-Soares

Cardoso

et al. 2020

Appl Microbiol Biotechnol

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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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Performance evaluation of anti-fixed Leishmania infantum promastigotes immunoglobulin G (IgG) detected by flow cytometry as a diagnostic tool for visceral Leishmaniasis

Cited by 3 publications

References 38 publications

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

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

Recent advances and new strategies in Leishmaniasis diagnosis

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

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