Establishment and Application of an Indirect Enzyme-Linked Immunosorbent Assay for Measuring GPI-Anchored Protein 52 (P52) Antibodies in -Infected Dogs

Overview

Journal Animals (Basel)

Date 2022 May 14

PMID 35565622

Authors

Qin Liu

Xueyan Zhan

Dongfang Li

Junlong Zhao

Haiyong Wei

Heba Alzan

Lan He

Affiliations

Soon will be listed here.

Abstract

is a malaria-like protozoan that parasitizes the red blood cells of canids to cause babesiosis. Due to its high expression and essential function in the survival of parasites, the Glycosylphosphatidylinositol (GPI) anchor protein family is considered an excellent immunodiagnostic marker. Herein, we identified a novel GPI-anchored protein named as BgGPI52-WH with a size of 52 kDa; the recombinant BgGPI52-WH with high antigenicity and immunogenicity was used as a diagnostic antigen to establish a new iELISA method. The iELISA had a sensitivity of 1:400, and no cross-reaction with other apicomplexan parasites occurred. We further demonstrated that the degree of variation was less than 10% using the same samples from the same or different batches of an enzyme-labeled strip. It was found that the method was able to detect early infection (6 days after infection) in the sera of the -infected experimental dogs in which antibody response to rBgGPI52-WH was evaluated. Clinical sera from pet hospitals were further tested, and the average positive rate was about 11.41% (17/149). The results indicate that BgGPI52-WH is a reliable diagnostic antigen, and the new iELISA could be used as a practical method for the early diagnosis of .

References

Bono M, Mangold A, Baravalle M, Valentini B, Thompson C, Wilkowsky S . Efficiency of a recombinant MSA-2c-based ELISA to establish the persistence of antibodies in cattle vaccinated with Babesia bovis. Vet Parasitol. 2008; 157(3-4):203-10. DOI: 10.1016/j.vetpar.2008.07.025. View

Casapulla R, Baldi L, Avallone V, Sannino R, Pazzanese L, Mizzoni V . Canine piroplasmosis due to Babesia gibsoni: clinical and morphological aspects. Vet Rec. 1998; 142(7):168-9. DOI: 10.1136/vr.142.7.168. View

Schares G, Rauser M, Sondgen P, Rehberg P, Barwald A, Dubey J . Use of purified tachyzoite surface antigen p38 in an ELISA to diagnose bovine neosporosis. Int J Parasitol. 2000; 30(10):1123-30. DOI: 10.1016/s0020-7519(00)00092-8. View

Singh M, Raina O, Sankar M, Rialch A, Tigga M, Kumar G . Molecular detection and genetic diversity of Babesia gibsoni in dogs in India. Infect Genet Evol. 2016; 41:100-106. DOI: 10.1016/j.meegid.2016.03.025. View

Wieser S, Schnittger L, Florin-Christensen M, Delbecq S, Schetters T . Vaccination against babesiosis using recombinant GPI-anchored proteins. Int J Parasitol. 2019; 49(2):175-181. DOI: 10.1016/j.ijpara.2018.12.002. View

Atkinson R, Harper P, Reichel M, Ellis J . Progress in the serodiagnosis of Neospora caninum infections of cattle. Parasitol Today. 2000; 16(3):110-4. DOI: 10.1016/s0169-4758(99)01604-x. View

Wahlgren M, Goel S, Akhouri R . Variant surface antigens of Plasmodium falciparum and their roles in severe malaria. Nat Rev Microbiol. 2017; 15(8):479-491. DOI: 10.1038/nrmicro.2017.47. View

Lekutis C, Ferguson D, Grigg M, Camps M, Boothroyd J . Surface antigens of Toxoplasma gondii: variations on a theme. Int J Parasitol. 2001; 31(12):1285-92. DOI: 10.1016/s0020-7519(01)00261-2. View

Man S, Fu Y, Guan Y, Feng M, Qiao K, Li X . Evaluation of a Major Surface Antigen of Merozoites as a Vaccine Candidate against Babesia Infection. Front Microbiol. 2018; 8:2545. PMC: 5742146. DOI: 10.3389/fmicb.2017.02545. View

10.

Karasova M, Tothova C, Grelova S, Fialkovicova M . The Etiology, Incidence, Pathogenesis, Diagnostics, and Treatment of Canine Babesiosis Caused by Infection. Animals (Basel). 2022; 12(6). PMC: 8944684. DOI: 10.3390/ani12060739. View

11.

Wozniak E, Barr B, Thomford J, Yamane I, McDonough S, Moore P . Clinical, anatomic, and immunopathologic characterization of Babesia gibsoni infection in the domestic dog (Canis familiaris). J Parasitol. 1997; 83(4):692-9. View

12.

Orlean P, Menon A . Thematic review series: lipid posttranslational modifications. GPI anchoring of protein in yeast and mammalian cells, or: how we learned to stop worrying and love glycophospholipids. J Lipid Res. 2007; 48(5):993-1011. DOI: 10.1194/jlr.R700002-JLR200. View

13.

Hafalla J, Silvie O, Matuschewski K . Cell biology and immunology of malaria. Immunol Rev. 2011; 240(1):297-316. DOI: 10.1111/j.1600-065X.2010.00988.x. View

14.

Xu J, Wang J, Li Z, He X, Zhao S, Ma Q . A universal ELISA assay for detecting six strains of ovine Babesia species in China. Vet Parasitol. 2021; 300:109616. DOI: 10.1016/j.vetpar.2021.109616. View

15.

Goo Y, Aboge G, Terkawi M, Jia H, Yamagishi J, Sunaga F . Four promising antigens, BgP32, BgP45, BgP47, and BgP50, for serodiagnosis of Babesia gibsoni infection were classified as B. gibsoni merozoite surface protein family. Parasitol Int. 2011; 61(2):364-8. DOI: 10.1016/j.parint.2011.11.007. View

16.

Hung C, Yu J, Seshan K, Reichard U, Cole G . A parasitic phase-specific adhesin of Coccidioides immitis contributes to the virulence of this respiratory Fungal pathogen. Infect Immun. 2002; 70(7):3443-56. PMC: 128074. DOI: 10.1128/IAI.70.7.3443-3456.2002. View

17.

Suzuki K, Kasai R, Hirosawa K, Nemoto Y, Ishibashi M, Miwa Y . Transient GPI-anchored protein homodimers are units for raft organization and function. Nat Chem Biol. 2012; 8(9):774-83. DOI: 10.1038/nchembio.1028. View

18.

Goo Y, Jia H, Aboge G, Terkawi M, Kuriki K, Nakamura C . Babesia gibsoni: Serodiagnosis of infection in dogs by an enzyme-linked immunosorbent assay with recombinant BgTRAP. Exp Parasitol. 2007; 118(4):555-60. DOI: 10.1016/j.exppara.2007.11.010. View

19.

Lequin R . Enzyme immunoassay (EIA)/enzyme-linked immunosorbent assay (ELISA). Clin Chem. 2005; 51(12):2415-8. DOI: 10.1373/clinchem.2005.051532. View

20.

Florin-Christensen M, Schnittger L, Dominguez M, Mesplet M, Rodriguez A, Ferreri L . Search for Babesia bovis vaccine candidates. Parassitologia. 2007; 49 Suppl 1:9-12. View