Skip to main content Accessibility help
×
Home

Use of polyclonal IgY antibodies to detect serum immune complexes in patients with active hookworm infection

  • Dayane Costa de Souza (a1), Lucas Silva de Faria (a1), José Eduardo Neto de Sousa (a1), Camila de Almeida Lopes (a1), Vanessa da Silva Ribeiro (a1), Vivian Jordania da Silva (a2), Raphaella Paula Ribeiro (a3), Élida Mara Leite Rabelo (a2), Veridiana Melo Rodrigues Ávila (a4), Álvaro Ferreira Júnior (a3) and Julia Maria Costa-Cruz (a1)...

Abstract

Definitive diagnosis of hookworm infection is usually based on the microscopic detection of eggs in a stool sample; however, several cases display a low or irregular egg output. Serodiagnosis can be a useful tool to identify these cases, but conventional tests do not differentiate past from active infections. The aim of this study was to obtain and apply egg yolk polyclonal immunoglobulin (IgY) antibodies to detect immune complexes (ICs) in serum samples from patients infected with hookworm. Hens were immunized with Ancylostoma ceylanicum saline extract, their eggs were collected and then IgY antibodies were extracted and purified. Antibody purity was tested by 12% sodium dodecyl sulphate polyacrylamide gel electrophoresis and specificity was assessed by immunoblotting and immunofluorescence. IgY production was evaluated by kinetics enzyme-linked immunosorbent assay (ELISA). Sandwich ELISA tested the ability of IgY to detect ICs in serum samples, from which diagnostic parameters were calculated. Antibody responses increased steadily from day 7 to 42. In the immunoblotting assay, IgY recognized two protein complexes. The immunofluorescence assay showed no staining in control samples. The sandwich ELISA presented a very high diagnostic value, with a sensitivity of 90% and a specificity of 86.7%. Our pioneer strategy highlights the potential use of egg yolk IgY as a diagnostic test to detect active hookworm infection.

Copyright

Corresponding author

Author for correspondence: Julia Maria Costa-Cruz, E-mail: costacruz@ufu.br

References

Hide All
Akita, EM and Nakai, S (1993) Comparison of four purification methods for the production of immunoglobulins from eggs laid by hens immunized with an enterotoxigenic E. coli strain. Journal of Immunological Methods 160, 207214.
Amro, WA, Al-Qaisi, W and Al-Razem, F (2018) Production and purification of IgY antibodies from chicken egg yolk. Journal of Genetic Engineering and Biotechnology 16, 99103.
Baermann, G (1917) Eine Einfache Methode zur Auffindung von Ankylostomum (Nematoden) Larven in Erdproben. In Mededeel. Mith H. Geneesk. Batavia. Lab Weltevreden Feestbundel 57, 4147.
Elsemore, DA, Geng, J, Cote, J, Hanna, R, Lucio-Forster, A and Bowman, DD (2017) Enzyme-linked immunosorbent assays for coproantigen detection of Ancylostoma caninum and Toxocara canis in dogs and Toxocara cati in cats. Journal of Veterinary Diagnostic Investigation 29, 645653.
Esmailnejad, A, Abdi-Hachesoo, B, Hosseini Nasab, E and Shakoori, M (2019) Production, purification, and evaluation of quail immunoglobulin Y against Salmonella typhimurium and Salmonella enteritidis. Molecular Immunology 107, 7983.
Faria, LS, de Souza, DLN, Ribeiro, RP, de Sousa, JEN, Borges, IP, Ávila, VMR, Ferreira-Júnior, Á, Goulart, LR and Costa-Cruz, JM (2019) Highly specific and sensitive anti-Strongyloides venezuelensis Igy antibodies applied to the human strongyloidiasis immunodiagnosis. Parasitology International 72, 101933.
Feliciano, ND, Gonzaga, HT, Gonçalves-Pires, MdR, Gonçalves, AL, Rodrigues, RM, Ueta, MT and Costa-Cruz, JM (2010) Hydrophobic fractions from Strongyloides venezuelensis for use in the human immunodiagnosis of strongyloidiasis. Diagnostic Microbiology and Infectious Diseases 67, 153161.
Ferreira Júnior, Á, Santiago, FM, Silva, MV, Ferreira, FB, Macêdo Júnior, AG, Mota, CM, Faria, MS, Silva Filho, HH, Silva, DA, Cunha-Júnior, JP, Mineo, JR and Mineo, TW (2012) Production, characterization and applications for Toxoplasma gondii-specific polyclonal chicken egg yolk immunoglobulins. PLoS ONE 7, e40391.
Friedman, RD (1982) Comparison of four different silver-staining techniques for salivary protein detection in alkaline polyacrylamide gels. Analytical Biochemistry 126, 346349.
George, S, Levecke, B, Kattula, D, Velusamy, V, Roy, S, Geldhof, P, Sarkar, R and Kang, G (2016) Molecular identification of hookworm isolates in humans, dogs and soil in a tribal area in Tamil Nadu, India. PLoS Neglected Tropical Diseases 10, e0004891.
Gonçalves, AL, Nunes, DS, Gonçalves-Pires, MR, Ueta, MT and Costa-Cruz, JM (2012) Use of larval, parasitic female and egg antigens from Strongyloides venezuelensis to detect parasite-specific IgG and immune complexes in immunodiagnosis of human strongyloidiasis. Parasitology 139, 956961.
Gonçalves, AL, de Araújo, KC, Carvalho, EF, Ueta, MT and Costa-Cruz, JM (2016) Specific IgG and immune complex responses to parthenogenetic females and eggs of nematode Strongyloides venezuelensis for the diagnosis of immunosuppression in infected rats. Journal of Helminthology 90, 342346.
Gonzaga, HT, Vila-Verde, C, Nunes, DS, Ribeiro, VS, Cunha-Júnior, JP and Costa-Cruz, JM (2013) Ion-exchange protocol to obtain antigenic fractions with potential for serodiagnosis of strongyloidiasis. Parasitology 140, 6975.
Hu, Y, Ellis, BL, Yiu, YY, Miller, MM, Urban, JF, Shi, LZ and Aroian, RV (2013) An extensive comparison of the effect of anthelmintic classes on diverse nematodes. PLoS ONE 8, e70702.
Laemmli, UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680685.
Larsson, A, Wejåker, PE, Forsberg, PO and Lindahl, T (1992) Chicken antibodies: a tool to avoid interference by complement activation in ELISA. Journal of Immunology Methods 156, 7983.
Lei, JH, Guan, F, Xu, H, Chen, L, Su, BT, Zhou, Y, Wang, T, Li, YL and Liu, WQ (2012) Application of an immunomagnetic bead ELISA based on IgY for detection of circulating antigen in urine of mice infected with Schistosoma japonicum. Veterinary Parasitology 187, 196202.
Leslie, GA and Clem, LW (1969) Phylogeny of immunoglobulin structure and function 3. Immunoglobulins of the chicken. Journal of Experimental Medicine 130, 13371352.
Li, X, Wang, L, Zhen, Y, Li, S and Xu, Y (2015) Chicken egg yolk antibodies (IgY) as non-antibiotic production enhancers for use in swine production: a review. Journal of Animal Science and Biotechnology 26, 40.
Liu, LN, Jing, FJ, Cui, J, Fu, GY and Wang, ZQ (2013) Detection of circulating antigen in serum of mice infected with Trichinella spiralis by an IgY-IgM mAb sandwich ELISA. Experimental Parasitology 133, 150155.
Lopes, CA, de Faria, LS, de Sousa, JEN, Borges, IP, Ribeiro, RP, Bueno, LL, Rodrigues Ávila, VM, Ferreira-Júnior, Á and Costa-Cruz, JM (2019) Anti-Ascaris suum immunoglobulin Y as a novel biotechnological tool for the diagnosis of human ascariasis. Journal of Helminthology 94, e71.
Lowry, OH, Rosebrough, NJ, Farr, AL and Randall, RJ (1951) Protein measurement with the Folin phenol reagent. The Journal of Biological Chemistry 193, 265275.
Lutz, A (1919) O Schistosomum mansoni e a schistosomatose segundo observaçoões feitas no Brasil. Memórias do Instituto Oswaldo Cruz 11, 121125.
Moraes, RG (1948) Contribuição para o estudo do Strongyloides Stercoralis e da estrongiloidíase no Brasil. Revista do Serviço Especial de Saúde Pública 1, 507624.
Nie, G, Wang, T, Lu, S, Liu, W, Li, Y and Lei, J (2014) Detection of Clonorchis sinensis circulating antigen in sera from Chinese patients by immunomagnetic bead ELISA based on IgY. PLoS ONE 9, e113208.
Nikolay, B, Brooker, SJ and Pullan, RL (2014) Sensitivity of diagnostic tests for human soil-transmitted helminth infections: a meta-analysis in the absence of a true gold standard. International Journal for Parasitology 44, 765774.
Ohnishi, T, Kakimoto, K, Hashida, S, Fujii, M, Hirono, S, Nishiyama, K, Amita, Y, Ishikawa, E, Tsubouchi, H and Daikuhara, Y (2000) Development of highly sensitive enzyme-linked immunosorbent assays for hepatocyte growth factor/scatter factor (HGF/SF): determination of HGF/SF in serum and urine from normal human subjects. Journal of Immunology Methods 244, 163173.
Ohyama, K, Huy, NT, Yoshimi, H, Kishikawa, N, Nishizawa, JE, Roca, Y, Revolvo Guzmán, RJ, Velarde, FU, Kuroda, N and Hirayama, K (2016) Proteomic profile of circulating immune complexes in chronic Chagas disease. Parasite Immunology 38, 609617.
Oliveira, HB, Machado, GA, Cabral, DD and Costa-Cruz, JM (2007) Application of Taenia saginata Metacestodes as an alternative antigen for the serological diagnosis of human neurocysticercosis. Parasitology Research 101, 10071013.
Prieto-Pérez, L, Pérez-Tanoira, R, Cabello-Úbeda, A, Petkova-Saiz, E and Górgolas-Hernandéz-Mora, M (2016) Geohelminths. Enfermedades Infecciosas y Microbiología Clínica 34, 384389.
Pullan, RL, Smith, JL, Jasrasaria, R and Brooker, SJ (2014) Global numbers of infection and disease burden of soil transmitted helminth infections in 2010. Parasites and Vectors 7, 37.
Ribeiro, VdS, Araújo, TG, Gonzaga, HT, Nascimento, R, Goulart, LR and Costa-Cruz, JM (2013) Development of specific scFv antibodies to detect neurocysticercosis antigens and potential applications in immunodiagnosis. Immunology Letters 156, 5967.
Rojko, JL, Evans, MG, Price, SA, Han, B, Waine, G, DeWitte, M, Haynes, J, Freimark, B, Martin, P, Raymond, JT, Evering, W, Rebelatto, MC, Schenck, E and Horvath, C (2014) Formation, clearance, deposition, pathogenicity, and identification of biopharmaceutical-related immune complexes: review and case studies. Toxicologic Pathology 42, 725764.
Schwarzkopf, C, Staak, C, Behn, I and Erhard, M (2001) Immunisation. In Schade, R, Behn, I, Erhard, M, Hlinak, A and Staak, C (eds), Chickens Egg Yolk Antibodies, Production and Applications-IgY-Technology. Berlin, Heidelberg: Springer-Verlag, pp. 2564.
Sudré, AP, Macedo, HW, Peralta, RHS and Peralta, JM (2006) Diagnóstico da estrongiloidíase humana: importância e técnicas. Revista de Patologia Tropical 35, 173184.
Thirumalai, D, Visaga Ambi, S, Vieira-Pires, RS, Xiaoying, Z, Sekaran, S and Krishnan, U (2019) Chicken egg yolk antibody (IgY) as diagnostics and therapeutics in parasitic infections – A review. International Journal of Biological Macromolecules 136, 755763.
Towbin, H, Staehelin, T and Gordon, J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America 76, 43504354.
Truscott, JE, Turner, HC, Farrell, SH and Anderson, RM (2016) Soil-transmitted helminths: mathematical models of transmission, the impact of mass drug administration and transmission elimination criteria. Advances in Parasitology 94, 133198.
Werkman, M, Wright, JE, Truscott, JE, Easton, AV, Oliveira, RG, Toor, J, Ower, A, Ásbjörnsdóttir, KH, Means, AR, Farrell, SH, Walson, JL and Anderson, RM (2018) Testing for soil-transmitted helminth transmission elimination: analysing the impact of the sensitivity of different diagnostic tools. PLoS Neglected Tropical Diseases 12, e0006114.
Zhang, H, Mi, T, Khan, OY, Sheng, Y, Eremin, SA, Beier, RC, Zhang, S, Shen, J and Wang, Z (2015) Fluorescence polarization immunoassay using IgY antibodies for detection of valnemulin in swine tissue. Analytical and Bioanalytical Chemistry 407, 78437848.

Keywords

Use of polyclonal IgY antibodies to detect serum immune complexes in patients with active hookworm infection

  • Dayane Costa de Souza (a1), Lucas Silva de Faria (a1), José Eduardo Neto de Sousa (a1), Camila de Almeida Lopes (a1), Vanessa da Silva Ribeiro (a1), Vivian Jordania da Silva (a2), Raphaella Paula Ribeiro (a3), Élida Mara Leite Rabelo (a2), Veridiana Melo Rodrigues Ávila (a4), Álvaro Ferreira Júnior (a3) and Julia Maria Costa-Cruz (a1)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.