An assessment of serological techniques for the identification of asymptomatic visceral leishmaniasis in blood donors in Northeastern Brazil

Authors

DOI:

https://doi.org/10.33448/rsd-v11i6.28827

Keywords:

Flow cytometry; Direct agglutination test; ELISA; Recombinant ELISA.

Abstract

Background: Visceral leishmaniasis (VL) is a neglected disease caused by species of the Leishmania donovani complex and transmitted mainly by the bite of infected sandflies. A less frequent form of transmission is the transfusion route, important in countries lacking screening procedures for VL in blood banks. Methods: To identify asymptomatic infection by Leishmania sp, our goal here was to evaluate and compare the performance of three different serological techniques: Flow Cytometry (FC); Direct Agglutination Test (DAT); and Immunoenzymatic Test (ELISA). Thirty-one asymptomatic carriers of L. infantum were selected, from a single blood bank located in Recife, northeastern Brazil. Results : Foi avaliada a frequência de amostras positivas assintomáticas nos testes, no ELISA baseado no antígeno solúvel de Leishmania (25,8%), FC (22,8%), DAT (9,7%) e ELISA baseado em uma única proteína recombinante (12,9%). A sensibilidade dos testes também foi avaliada, o FC (22,58%) e o ELISA com o antígeno solúvel (22,58%) apresentaram os maiores valores de sensibilidade que o DAT (6,45%) e o ELISA recombinante (12,9%). Apenas a CF apresentou baixa especificidade (22,22%), em contraste com as demais técnicas (100% de especificidade). Conclusion: Given the results presented, we highlight the low effectiveness of serological techniques for the identification of asymptomatic individuals, thus emphasizing the need to develop more accurate methods for this purpose.

References

Machado de Assis, T. S., Azeredo-da-Silva, A. L., Werneck, G. L., & Rabello, A. (2016). Cost-effectiveness analysis of diagnostic tests for human visceral leishmaniasis in Brazil. Transactions of the Royal Society of Tropical Medicine and Hygiene, 110(8), 464–471.

Brazil. (2019). Ministério da Saúde. Guia de vigilância epidemiológica. Normas e Manuais Técnicos. (3a ed.), Ministério da Saúde.

Caballero, Z. C., Sousa, O. E., Marques, W. P., Saez-Alquezar, A., & Umezawa, E. S. (2007). Evaluation of serological tests to identify Trypanosoma cruzi infection in humans and determine cross-reactivity with Trypanosoma rangeli and Leishmania spp. Clinical and vaccine immunology : CVI, 14(8), 1045–1049.

de Carvalho, F., Riboldi, E. O., Bello, G. L., Ramos, R. R., Barcellos, R. B., Gehlen, M., Halon, M. L., Romão, P., Dallegrave, E., & Rossetti, M. (2018). Canine visceral leishmaniasis diagnosis: a comparative performance of serological and molecular tests in symptomatic and asymptomatic dogs. Epidemiology and infection, 146(5), 571–576..

Chappuis, F., Rijal, S., Soto, A., Menten, J., & Boelaert, M. (2006). A meta-analysis of the diagnostic performance of the direct agglutination test and rK39 dipstick for visceral leishmaniasis. BMJ (Clinical research ed.), 333(7571), 723.

Cota, G. F., de Sousa, M. R., de Freitas Nogueira, B. M., Gomes, L. I., Oliveira, E., Assis, T. S., de Mendonça, A. L., Pinto, B. F., Saliba, J. W., & Rabello, A. (2013). Comparison of parasitological, serological, and molecular tests for visceral leishmaniasis in HIV-infected patients: a cross-sectional delayed-type study. The American journal of tropical medicine and hygiene, 89(3), 570–577.

ElmahallawyE. K., Sampedro MartinezA., Rodriguez-GrangerJ., Hoyos-MallecotY., AgilA., Navarro MariJ. M., & Gutierrez FernandezJ. (2014). Diagnosis of leishmaniasis. The Journal of Infection in Developing Countries, 8(08), 961-972.

Ferreira-Silva, M. M., Teixeira, L., Tibúrcio, M. S., Pereira, G. A., Rodrigues, V., Palis, M., Afonso, P., Alves, M., Feitosa, J. M., Urias, E., Santos, E. M., Carvalho, S., & Moraes-Souza, H. (2018). Socio-epidemiological characterisation of blood donors with asymptomatic Leishmania infantum infection from three Brazilian endemic regions and analysis of the transfusional transmission risk of visceral leishmaniasis. Transfusion medicine (Oxford, England), 28(6), 433–439.

França, A., de Castro, V. L., Lima, M. S., Jr, Pontes, E. R., & Dorval, M. E. (2013). Anti-Leishmania antibodies in blood donors from the Midwest region of Brazil. Transfusion and apheresis science : official journal of the World Apheresis Association : official journal of the European Society for Haemapheresis, 49(3), 627–630.

Fukutani, K. F., Figueiredo, V., Celes, F. S., Cristal, J. R., Barral, A., Barral-Netto, M., & de Oliveira, C. I. (2014). Serological survey of Leishmania infection in blood donors in Salvador, Northeastern Brazil. BMC infectious diseases, 14, 422.

Gadisa, E., Custodio, E., Cañavate, C., Sordo, L., Abebe, Z., Nieto, J., Chicharro, C., Aseffa, A., Yamuah, L., Engers, H., Moreno, J., & Cruz, I. (2012). Usefulness of the rK39-immunochromatographic test, direct agglutination test, and leishmanin skin test for detecting asymptomatic Leishmania infection in children in a new visceral leishmaniasis focus in Amhara State, Ethiopia. The American journal of tropical medicine and hygiene, 86(5), 792–798.

Galluzzi, L., Ceccarelli, M., Diotallevi, A., Menotta, M., & Magnani, M. (2018). Real-time PCR applications for diagnosis of leishmaniasis. Parasites & vectors, 11(1), 273.

Gama Ker, H., Dian de Oliveira Aguiar-Soares, R., Mendes Roatt, B., das Dores Moreira, N., Coura-Vital, W., Martins Carneiro, C., Teixeira-Carvalho, A., Martins-Filho, O. A., Cordeiro Giunchetti, R., da Silveira-Lemos, D., & Barbosa Reis, A. (2013). Effect of the preservative and temperature conditions on the stability of Leishmania infantum promastigotes antigens applied in a flow cytometry diagnostic method for canine visceral leishmaniasis. Diagnostic microbiology and infectious disease, 76(4), 470–476.

Garcez, Lourdes M., Shaw, Jeffrey J. e Silveira, Fernando T. (1996). Teste de aglutinação direta no sorodiagnóstico da leishmaniose visceral no estado do Pará. Revista da Sociedade Brasileira de Medicina Tropical [online]. v. 29, n. 2 , pp. 165-180.

Gomes, I. T., Carvalho, S. F., Rocha, R. D., Peruhype-Magalhães, V., Dietze, R., Martins-Filho, O. A., & Lemos, E. M. (2010). Anti-Leishmania chagasi immunoglobulin G3 detected by flow cytometry for early cure assessment in American visceral leishmaniasis. Journal of immunological methods, 360(1-2), 76–83.

Gontijo, Célia Maria Ferreira e Melo, Maria Norma.(2004) Leishmaniose visceral no Brasil: quadro atual, desafios e perspectivas. Revista Brasileira de Epidemiologia [online]. v. 7, n. 3, pp. 338-349.

Harith, A. E., Kolk, A. H., Kager, P. A., Leeuwenburg, J., Muigai, R., Kiugu, S., Kiugu, S., & Laarman, J. J. (1986). A simple and economical direct agglutination test for serodiagnosis and sero-epidemiological studies of visceral leishmaniasis. Transactions of the Royal Society of Tropical Medicine and Hygiene, 80(4)

Ibarra-Meneses, A. V., Ghosh, P., Hossain, F., Chowdhury, R., Mondal, D., Alvar, J., Moreno, J., & Carrillo, E. (2017). IFN-γ, IL-2, IP-10, and MIG as Biomarkers of Exposure to Leishmania spp., and of Cure in Human Visceral Leishmaniasis. Frontiers in cellular and infection microbiology, 7, 200.

Ker, H. G., Coura-Vital, W., Valadares, D. G., Aguiar-Soares, R., de Brito, R., Veras, P., Fraga, D., Martins-Filho, O. A., Teixeira-Carvalho, A., & Reis, A. B. (2019). Multiplex flow cytometry serology to diagnosis of canine visceral leishmaniasis. Applied microbiology and biotechnology, 103(19), 8179–8190.

Kühne, V., Verstraete, R., van Ostade, X., & Büscher, P. (2020). Experimental Evidence on the Nature of the Antigen in the Direct Agglutination Test for Visceral Leishmaniasis. The American journal of tropical medicine and hygiene, 102(4), 788–796..

Landis, J. R., & Koch, G. G. (1977). The measurement of observer agreement for categorical data. Biometrics, 33(1), 159–174.

Liew, F, Y., O'donnell, C,A., Immunology of leishmaniasis. In: Advances in Parasitology. Academic Press, 1993; 161–259.

de Lima, V. M., Fattori, K. R., Michelin, A., da Silveira Neto, L., & Vasconcelos, R. (2010). Comparison between ELISA using total antigen and immunochromatography with antigen rK39 in the diagnosis of canine visceral leishmaniasis. Veterinary parasitology, 173(3-4), 330–333.

McKinnon K. M. (2018). Flow Cytometry: An Overview. Current protocols in immunology, 120, 5.1.1–5.1.11. https://doi.org/10.1002/cpim.40

Mettler, M., Grimm, F., Capelli, G., Camp, H., & Deplazes, P. (2005). Evaluation of enzyme-linked immunosorbent assays, an immunofluorescent-antibody test, and two rapid tests (immunochromatographic-dipstick and gel tests) for serological diagnosis of symptomatic and asymptomatic Leishmania infections in dogs. Journal of clinical microbiology, 43(11), 5515–5519.

Michel, G., Pomares, C., Ferrua, B., & Marty, P. (2011). Importance of worldwide asymptomatic carriers of Leishmania infantum (L. chagasi) in human. Acta tropica, 119(2-3), 69–75.

Mohammed, E. A., Wright, E. P., Kager, P. A., Laarman, J. J., & Pondman, K. W. (1985). ELISA using intact promastigotes for immunodiagnosis of kala-azar. Transactions of the Royal Society of Tropical Medicine and Hygiene, 79(3), 344–350.

Nolan, J. P., & Condello, D. (2013). Spectral flow cytometry. Current protocols in cytometry, Chapter 1, Unit1.27. https://doi.org/10.1002/0471142956.cy0127s63

Oliveira, G. G., Magalhães, F. B., Teixeira, M. C., Pereira, A. M., Pinheiro, C. G., Santos, L. R., Nascimento, M. B., Bedor, C. N., Albuquerque, A. L., dos-Santos, W. L., Gomes, Y. M., Moreira, E. D., Jr, Brito, M. E., Pontes de Carvalho, L. C., & de Melo Neto, O. P. (2011). Characterization of novel Leishmania infantum recombinant proteins encoded by genes from five families with distinct capacities for serodiagnosis of canine and human visceral leishmaniasis. The American journal of tropical medicine and hygiene, 85(6), 1025–1034.

Pedral-Sampaio, G., Alves, J. S., Schriefer, A., Magalhães, A., Meyer, R., Glesby, M. J., Carvalho, E. M., & Carvalho, L. P. (2016). Detection of IgG Anti-Leishmania Antigen by Flow Cytometry as a Diagnostic Test for Cutaneous Leishmaniasis. PloS one, 11(9), e0162793.

Porcino, G. N., Carvalho, K., Braz, D. C., Costa Silva, V., Costa, C., & de Miranda Santos, I. (2019). Evaluation of methods for detection of asymptomatic individuals infected with Leishmania infantum in the state of Piauí, Brazil. PLoS neglected tropical diseases, 13(7), e0007493.

Reithinger, R., Quinnell, R. J., Alexander, B., & Davies, C. R. (2002). Rapid detection of Leishmania infantum infection in dogs: comparative study using an immunochromatographic dipstick test, enzyme-linked immunosorbent assay, and PCR. Journal of clinical microbiology, 40(7), 2352–2356.

Riera, C., Fisa, R., López-Chejade, P., Serra, T., Girona, E., Jiménez, M., Muncunill, J., Sedeño, M., Mascaró, M., Udina, M., Gállego, M., Carrió, J., Forteza, A., & Portús, M. (2008). Asymptomatic infection by Leishmania infantum in blood donors from the Balearic Islands (Spain). Transfusion, 48(7), 1383–1389.

Riera, C., Fisa, R., Udina, M., Gállego, M., & Portus, M. (2004). Detection of Leishmania infantum cryptic infection in asymptomatic blood donors living in an endemic area (Eivissa, Balearic Islands, Spain) by different diagnostic methods. Transactions of the Royal Society of Tropical Medicine and Hygiene, 98(2), 102–110.

Rocha, R. D., Gontijo, C. M., Elói-Santos, S. M., Teixeira Carvalho, A., Corrêa-Oliveira, R., Marques, M. J., Genaro, O., Mayrink, W., & Martins-Filho, O. A. (2002). Anticorpos antipromastigotas vivas de Leishmania (Viannia) braziliensis, detectados pela citometria de fluxo, para identificação da infecção ativa na leishmaniose tegumentar americana [Anti-live Leishmania (Viannia) braziliensis promastigote antibodies, detected by flow cytometry, to identify active infection in american cutaneous leishmaniasis]. Revista da Sociedade Brasileira de Medicina Tropical, 35(6), 551–562.

Sakkas, H., Gartzonika, C., & Levidiotou, S. (2016). Laboratory diagnosis of human visceral leishmaniasis. Journal of vector borne diseases, 53(1), 8–16.

Santos, W., Tavares, D., Castro Neto, A. L., Nascimento, M. B., Dhalia, R., Albuquerque, A. L., Costa, C., Magalhães, F. B., Rezende, A. M., & de Melo Neto, O. P. (2020). Gene design, optimization of protein expression and preliminary evaluation of a new chimeric protein for the serological diagnosis of both human and canine visceral leishmaniasis. PLoS neglected tropical diseases, 14(7), e0008488.

Silva, E. D., Oliveira, B. C., Oliveira, A. P., Santos, W., Diniz, G. T., de Melo Neto, O. P., Costa, C., Silva, M., Andrade, L. D., Medeiros, Z. M., & Pereira, V. (2019). Performance evaluation of anti-fixed Leishmania infantum promastigotes immunoglobulin G (IgG) detected by flow cytometry as a diagnostic tool for visceral Leishmaniasis. Journal of immunological methods, 469, 18–25.

Silva, L. P., Montenegro, S., Werkauser, R., Sales, K., Soares, F., Costa, V., Bezerra, A. C., Pinto, M., Ferreira, S. M., Neitzke-Abreu, H. C., Dantas-Torres, F., & Lima Junior, M. (2020). Asymptomatic Leishmania infection in blood donors from a major blood bank in Northeastern Brazil: a cross-sectional study. Revista do Instituto de Medicina Tropical de Sao Paulo, 62, e92.

Silvestre, R., Santarém, N., Cunha, J., Cardoso, L., Nieto, J., Carrillo, E., Moreno, J., & Cordeiro-da-Silva, A. (2008). Serological evaluation of experimentally infected dogs by LicTXNPx-ELISA and amastigote-flow cytometry. Veterinary parasitology, 158(1-2), 23–30.

Singh, S., & Sivakumar, R. (2003). Recent advances in the diagnosis of leishmaniasis. Journal of postgraduate medicine, 49(1), 55–60.

Srivastava, P., Dayama, A., Mehrotra, S., & Sundar, S. (2011). Diagnosis of visceral leishmaniasis. Transactions of the Royal Society of Tropical Medicine and Hygiene, 105(1), 1–6.

Srividya, G., Kulshrestha, A., Singh, R., & Salotra, P. (2012). Diagnosis of visceral leishmaniasis: developments over the last decade. Parasitology research, 110(3), 1065–1078.

Sundar, S., & Singh, O. P. (2018). Molecular Diagnosis of Visceral Leishmaniasis. Molecular diagnosis & therapy, 22(4), 443–457.

Terán-Angel, Guillermo, Schallig, Henk, Zerpa, Olga, Rodríguez, Vestalia, Ulrich, Marian, & Cabrera, Maira. (2007). Evaluación de una prueba de aglutinación directa como método alternativo para el diagnóstico de leishmaniasis visceral canina y humana en Venezuela. Biomédica, 27(3), 447-453.

Thakur, S., Joshi, J., & Kaur, S. (2020). Leishmaniasis diagnosis: an update on the use of parasitological, immunological and molecular methods. Journal of parasitic diseases : official organ of the Indian Society for Parasitology, 44(2), 253–272.

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23/04/2022

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SILVA, L. P.; SILVA, E. D. .; PEREIRA, A. M. de S. .; SANTOS, W. J. T. dos .; SILVA, A. H. da .; NEITZKE-ABREU, H. C. .; MEDEIROS, Z. M. de .; LIMA JÚNIOR, M. S. da C. .; PEREIRA, V. R. A. . An assessment of serological techniques for the identification of asymptomatic visceral leishmaniasis in blood donors in Northeastern Brazil. Research, Society and Development, [S. l.], v. 11, n. 6, p. e14011628827, 2022. DOI: 10.33448/rsd-v11i6.28827. Disponível em: https://rsdjournal.org/index.php/rsd/article/view/28827. Acesso em: 28 may. 2022.

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Health Sciences