Hostname: page-component-cd9895bd7-lnqnp Total loading time: 0 Render date: 2024-12-28T13:41:01.629Z Has data issue: false hasContentIssue false
  • English
  • Français

Evaluation of two heterologous recombinant antigens for the serological diagnosis of human polycystic echinococcosis

Published online by Cambridge University Press:  17 March 2022

D. Daipert-Garcia ,
V.G. Virginio ,
F.B. Oliveira ,
N.G. Siqueira ,
H.B. Ferreira  and
R. Rodrigues-Silva Open the ORCID record for R. Rodrigues-Silva [Opens in a new window]
D. Daipert-Garcia
Affiliation:
Laboratório de Helmintos Parasitos de Vertebrados, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, RJ, Brazil
V.G. Virginio
Affiliation:
Laboratório de Genômica Estrutural e Funcional, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
F.B. Oliveira
Affiliation:
Laboratório de Helmintos Parasitos de Vertebrados, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, RJ, Brazil
N.G. Siqueira
Affiliation:
Fundacão Hospital Estadual do Acre, Acre, Brazil Departamento de Medicina, Universidade Federal do Acre, Acre, Brazil
H.B. Ferreira
Affiliation:
Laboratório de Genômica Estrutural e Funcional, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
R. Rodrigues-Silva*
Affiliation:
Laboratório de Helmintos Parasitos de Vertebrados, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, RJ, Brazil
*
Author for correspondence: R. Rodrigues-Silva. E-mail: rosanrsilva@gmail.com
Get access Rights & Permissions [Opens in a new window]

Abstract

Polycystic echinococcosis (PE) is a zoonosis endemic in the Neotropical region of the Americas. It is caused by the larval stage of the cestode Echinococcus vogeli, which develops as harmful cysts that slowly grow in the liver, lungs and other organs of humans and other host species. Human PE diagnosis is usually based on clinical and epidemiological aspects and imaging techniques, often requiring confirmation by immunological assays. The currently available serological tests for detecting antibodies against Echinococcus spp. are mostly based on complex, variable and poorly characterized mixtures of native parasite antigens, which impairs specificity and/or sensitivity. In this scenario, the evaluation of well-characterized alternative antigens is urgently needed for the improvement of PE diagnosis. Here, two subunits (AgB8/1 and AgB8/2) of the major secretory antigen from Echinococcus granulosus (antigen B (AgB)), of diagnostic value for cystic echinococcosis, were validated for PE diagnosis. These antigens, produced as pure recombinant proteins (rAgB8/1 and rAgB8/2) in Escherichia coli, allowed detecting specific immunoglobulin G antibodies in sera from PE patients in an enzyme-linked immunosorbent assay, with sensitivities of 83.72% and 81.40%, respectively, and specificities of 83.12% and 80.09%, respectively. The use of recombinant proteins overcomes difficulties to obtain parasite material and reduced non-specific reactions and costs. Our results demonstrated reproducibility and accuracy high enough to be considered valid according to the acceptance criteria for Food and Drug Administration assay validation. This qualifies rAgB8/1 and rAgB8/2 as potential substitutes for the currently used parasite crude or partially purified antigens.

Keywords

Echinococcus vogelipolycystic echinococcosisantigen Brecombinant antigensELISAimmunodiagnosis
Type
Research Paper
Information
Journal of Helminthology , Volume 96 , 2022 , e21
Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Almeida, SCX, Martins, RLM, Moraes, MAP, Viegas, CA and Grilo, M (1997) Hidatidose pulmonar policística mimetizando lesões metastáticas: relato de caso. Jornal de Pneumologia 23, 261–263.Google Scholar
Amagai, M, Komai, A, Hashimoto, T, et al. (1999) Usefulness of enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3 for serodiagnosis of pemphigus. British Journal of Dermatology 140, 351–357.CrossRefGoogle ScholarPubMed
Brehm, K and Koziol, U (2017) Echinococcus-host interactions at cellular and molecular levels. Advances in Parasitology 95, 147–212.CrossRefGoogle ScholarPubMed
Chemale, G, Haag, KL, Ferreira, HB and Zaha, A (2001) Echinococcus granulosus antigen B is encoded by a gene family. Molecular and Biochemical Parasitology 116, 233–237.CrossRefGoogle ScholarPubMed
Craig, PS, Hegglin, D, Lightowlers, MW, Torgerson, PR and Wang, Q (2017) Echinococcosis: control and prevention. Advances in Parasitology 96, 55–158.CrossRefGoogle ScholarPubMed
D'Alessandro, A (1997) Polycystic echinococcosis in tropical America: Echinococcus vogeli and E. Oligarthrus. Acta Tropica 67, 43–65.CrossRefGoogle ScholarPubMed
D'Alessandro, A and Rausch, RL (2008) New aspects of neotropical polycystic (Echinococcus vogeli) and unicystic (Echinococcus oligarthrus) echinococcosis. Clinical Microbiology Reviews 21, 380–401.CrossRefGoogle ScholarPubMed
Debourgogne, A, Blanchet, D, Fior, A, Umhang, G, Simon, S and Aznar, C (2017) Neotropical echinococcosis caused by Echinococcus vogeli in a 6-year-old child: the second case report in humans in French Guiana. Paediatrics and International Child Health 37, 63–65.CrossRefGoogle Scholar
De la Rue, ML, Yamano, K, Almeida, CE, Lesbich, MP, Fernandes, CD, Goto, A, Kouguchi, H and Takahashi, K (2010) Serological reactivity of patients with Echinococcus infections (E. granulosus, E. Vogeli, and E. multilocularis) against three antigen B subunits. Parasitology Research 106, 741–745.CrossRefGoogle Scholar
Esfedan, AF, Sarkari, B and Mikaeili, F (2018) Genetic variability of antigen B8/1 among Echinococcus granulosus isolates from human, cattle, and sheep in Fars province, Southern Iran. Reports of Biochemistry and Molecular Biology 6, 164–169.Google Scholar
Faramarzi, T, Mobasheri, M, Yoosefy, A and Valadkhani, Z (2019) Expression and purification of truncated recombinant B8/1 protein of Echinococcus granulosus for diagnosis of hydatid infection in human. Acta Tropica 191, 139–145.CrossRefGoogle ScholarPubMed
Fernández, V, Ferreira, HB, Fernández, C, Zaha, A and Nieto, A (1996) Molecular characterisation of a novel 8-kDa subunit of Echinococcus granulosus antigen B. Molecular and Biochemical Parasitology 77, 247–250.CrossRefGoogle ScholarPubMed
Flores, MS, Carrillo, P, Tamez, E, Rangel, R, Rodríguez, EG, Maldonado, MG, Isibasi, A and Galán, L (2016) Diagnostic parameters of serological ELISA for invasive amoebiasis, using antigens preserved without enzymatic inhibitors. Experimental Parasitology 161, 48–53.CrossRefGoogle ScholarPubMed
Folle, AM, Kitano, ES, Lima, A, et al. (2017) Characterisation of antigen B protein species present in the hydatid cyst fluid of Echinococcus canadensis G7 genotype. PLOS Neglected Tropical Diseases 11, e0005250.CrossRefGoogle ScholarPubMed
Genzini, T, Siqueira, NG, Noujaim, HM, Santos, RG, Yamashita, ET, Trevizol, AP and Perosa, M (2013) Liver transplantation for neotropical polycystic echinococcosis caused by Echinococcus vogeli: a case report. Revista da Sociedade Brasileira de Medicina Tropical 46, 119–120.CrossRefGoogle ScholarPubMed
González-Sapienza, G, Lorenzo, C and Nieto, A (2000) Improved immunodiagnosis of cystic hydatid disease by using a synthetic peptide with higher diagnostic value than that of its parent protein, Echinococcus granulosus antigen B. Journal of Clinical Microbiology 38, 3979–3983.CrossRefGoogle ScholarPubMed
Greene, RM, Hancock, K, Wilkins, PP and Tsang, V (2000) Taenia solium: molecular cloning and serologic evaluation of 14 and 18 kDa related, diagnostic antigens. Journal of Parasitology 86, 1001–1007.CrossRefGoogle ScholarPubMed
Hanley, JA and McNeil, BJ (1982) The meaning and use of the area under a receiver operating characteristic (ROC) curve. Radiology 143, 29–36.CrossRefGoogle Scholar
Iraqi, W (2016) Diagnostic value of semi-purified antigens of hydatid cyst fluid in human cystic echinococcosis. Acta Parasitologica 61, 144–150.CrossRefGoogle ScholarPubMed
Jiang, L, Zhang, YG, Liu, MX and Feng, Z (2012) Analysis on the reactivity of five subunits of antigen B family in serodiagnosis of echinococcosis. Experimental Parasitology 131, 85–91.CrossRefGoogle ScholarPubMed
Johnstone, A and Thorpe, R (1982) Basic techniques. Immunochemistry in practice. Oxford, Blackwell Scientific Publications, 1–17Google Scholar
Kern, P, Silva, AM, Akhan, O, Müllhaupt, B, Vizcaychipi, KA, Budke, C and Vuitton, DA (2017) The echinococcoses: diagnosis, clinical management and burden of disease. Advances in Parasitology 96, 259–369.CrossRefGoogle ScholarPubMed
Li, J, Zhang, WB and McManus, DP (2004) Recombinant antigens for immunodiagnosis of cystic echinococcosis. Biological Procedures Online 6, 67–77.CrossRefGoogle ScholarPubMed
Lightowlers, MW, Liu, DY, Haralambous, A and Rickard, MD (1989) Subunit composition and specificity of the major cyst fluid antigens of Echinococcus granulosus. Molecular and Biochemical Parasitology 37, 171–182.CrossRefGoogle ScholarPubMed
Lorenzo, C, Ferreira, HB, Monteiro, KM, et al. (2005) Comparative analysis of the diagnostic performance of six major Echinococcus granulosus antigens assessed in a double-blind, randomized multicenter study. Journal of Clinical Microbiology 43, 2764–2770.CrossRefGoogle Scholar
Maddison, SE, Slemenda, SB, Schantz, PM, Fried, JA, Wilson, M and Tsang, VC (1989) A specific diagnostic antigen of Echinococcus granulosus with an apparent molecular weight of 8 kDa. American Journal of Tropical Medicine and Hygiene 40, 377–383.CrossRefGoogle ScholarPubMed
Manzano-Roman, R, Sanchez-Ovejero, C, Hernandez-Gonzalez, A, Casulli, A and Siles-Lucas, M (2015) Serological diagnosis and follow-up of human cystic echinococcosis: a new hope for the future? BioMed Research International 2015, 428205.CrossRefGoogle ScholarPubMed
Mihmanli, M, Idiz, UO, Kaya, C, Demir, U, Bostanci, O, Omeroglu, S and Bozkurt, E (2016) Current status of diagnosis and treatment of hepatic echinococcosis. World Journal of Hepatology 8, 1169–1181.CrossRefGoogle ScholarPubMed
Mohammadzadeh, T, Sako, Y, Sadjjadi, SM, Sarkari, B and Ito, A (2012) Comparison of the usefulness of hydatid cyst fluid, native antigen B and recombinant antigen B8/1 for serological diagnosis of cystic echinococcosis. Transactions of the Royal Society of Tropical Medicine and Hygiene 106, 371–375.CrossRefGoogle ScholarPubMed
Monteiro, KM, Zaha, A and Ferreira, HB (2008) Recombinant subunits as tools for the structural and functional characterization of Echinococcus granulosus antigen B. Experimental Parasitology 119, 490–498.CrossRefGoogle ScholarPubMed
Monteiro, KM, Cardoso, MB, Follmer, C, da Silveira, NP, Vargas, DM, Kitajima, EW, Zaha, A and Ferreira, HB (2012) Echinococcus granulosus antigen B structure: subunit composition and oligomeric states. PLOS Neglected Tropical Diseases 6, e1551.CrossRefGoogle ScholarPubMed
Mubanga, C, Mwape, KE, Phiri, IK, et al. (2019) Progress on the development of rapid diagnostic tests for foodborne neglected zoonotic helminthiases: a systematic review. Acta Tropica 194, 135–147.CrossRefGoogle ScholarPubMed
Novo Alonso, C, Ferreira, JG, Fernández, AR, Antequera, EH, Blanco, JAZ and Gajate, GV (1993) Hepatic alveolar hydatidosis. Revista Española de Enfermedades Digestivas 84, 127–129.Google ScholarPubMed
Obregón-Henao, A, Gil, DL, Gómez, DI, Sanzón, F, Teale, JM and Restrepo, BI (2001) The role of N-linked carbohydrates in the antigenicity of Taenia solium metacestode glycoproteins of 12, 16 and 18 kD. Molecular and Biochemical Parasitology 114, 209–215.CrossRefGoogle ScholarPubMed
Pagnozzi, D, Tamarozzi, F, Roggio, AM, et al. (2018) Structural and immunodiagnostic characterization of synthetic antigen B subunits from Echinococcus granulosus and their evaluation as target antigens for cyst viability assessment. Clinical Infectious Diseases 66, 1342–1351.CrossRefGoogle ScholarPubMed
Pedrosa, I, Saíz, A, Arrazola, J, Ferreirós, J and Pedrosa, CS (2000) Hydatid disease: radiological and pathological features and complications. Radiographics 20, 795–817.CrossRefGoogle Scholar
Petrone, L, Tamarozzi, F, Vola, A, Morales, MAG, Ludovisi, A, Fard, SN, Mariconti, M, Brunetti, E and Goletti, DI (2021) Accuracy of an experimental whole-blood test for detecting reactivation of echinococcal cysts. PloS Neglected Tropical Diseases 15(8), e0009648.CrossRefGoogle ScholarPubMed
Robles, Y, González, E, Govezensky, T, Mungia, ME, Acero, G, Bobes, RJ, Gevorkian, G and Manoutcharian, K (2005) Isolation of the Taenia crassiceps antigens from a phage display cDNA library and evaluation of their use for diagnosis of neurocysticercosis. Clinical Immunology 116, 265–270.CrossRefGoogle ScholarPubMed
Romig, T, Deplazes, P, Jenkins, D, Giraudoux, P, Massolo, A, Craig, PS, Wassermann, M, Takahashi, K and de la Rue, M (2017) Ecology and life cycle patterns of Echinococcus species. Advances in Parasitology 95, 213–314.CrossRefGoogle ScholarPubMed
Rott, MB, Fernández, V, Farias, S, Ceni, J, Ferreira, HB, Haag, KL and Zaha, A (2000) Comparative analysis of two different subunits of antigen B from Echinococcus granulosus: gene sequences, expression in Escherichia coli and serological evaluation. Acta Tropica 75(3), 331–340.CrossRefGoogle ScholarPubMed
Sarkari, B and Rezaei, Z (2015) Immunodiagnosis of human hydatid disease: where do we stand? World Journal of Methodology 5, 185–195.CrossRefGoogle ScholarPubMed
Savardashtaki, A, Sarkari, B, Arianfar, F and Mostafavi, Z (2017) Immunodiagnostic value of Echinococcus granulosus recombinant B8/1 subunit of antigen B. Iranian Journal of Immunology 14, 111–122.Google ScholarPubMed
Savardashtaki, A, Mostafavi-Pour, Z, Arianfar, F and Sarkari, B (2019) Comparison of the utility of recombinant B8/2 subunit of the antigen B, native antigen, and a commercial ELISA kit in the diagnosis of human cystic echinococcosis. Iranian Biomedical Journal 23, 246–252.Google Scholar
Siles-Lucas, M, Casulli, A, Conraths, FJ and Müller, N (2017) Laboratory diagnosis of Echinococcus spp. in human patients and infected animals. Advances in Parasitology 96, 159–257.CrossRefGoogle ScholarPubMed
Silva, CAM, Soares, MCP, Alves, MM, Conde, SRS, Amaral, IA and Teixeira, LSC (2002) Hidatidose/equinococose na amazônia oriental brasileira – atualização da casuística. Revista da Sociedade Brasileira de Medicina Tropical 35, 243.Google Scholar
Silva, ED, Cancela, M, Monteiro, KM, Ferreira, HB and Zaha, A (2018) Antigen B from Echinococcus granulosus enters mammalian cells by endocytic pathways. PLOS Neglected Tropical Diseases 12, e0006473.CrossRefGoogle ScholarPubMed
Siqueira, NG, Siqueira, CMVM, Rodrigues-Silva, R, Soares, MCP and Póvoa, MM (2013) Polycystic echinococcosis in the state of Acre, Brazil: contribution to patient diagnosis, treatment and prognosis. Memórias do Instituto Oswaldo Cruz 108, 533–540.CrossRefGoogle Scholar
Smith, DB and Johnson, KS (1998) Single-step purification polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67, 31–40.CrossRefGoogle Scholar
Soares, MC, Moreira-Silva, CA, Alves, MM, et al. (2004) Equinococose policística na amazônia oriental brasileira: Atualização da casuística. Revista da Sociedade Brasileira de Medicina Tropical 37, 75–83.CrossRefGoogle Scholar
Thompson, RCA (2017) Biology and systematics of Echinococcus. Advances in Parasitology 95, 65–109.CrossRefGoogle ScholarPubMed
USDHHS, FDA, CDER. Guidance for Industry: Bioanalytical Method Validation. US Department of Health and Human Services, Food and Drug Administration, 2001.Google Scholar
Virginio, VG, Hernández, A, Rott, MB, Monteiro, KM, Zandonai, AF, Nieto, A, Zaha, A and Ferreira, HB (2003) A set of recombinant antigens from Echinococcus granulosus with potential for use in the immunodiagnosis of human cystic hydatid disease. Clinical & Experimental Immunology 132, 309–315.CrossRefGoogle ScholarPubMed
Wang, N, Zhu, H, Zhan, J, et al. (2019) Cloning, expression, characterization, and immunological properties of citrate synthase from Echinococcus granulosus. Parasitology Research 118, 1811–1820.CrossRefGoogle ScholarPubMed
Wen, H, Vuitton, L, Tuxun, T, Li, J, Vuitton, DA, Zhang, W and McManus, DP (2019) Echinococcosis: advances in the 21st century. Clinical Microbiology Reviews 32, 1–39.CrossRefGoogle ScholarPubMed