Identification of a Cyclospora cayetanensis Oocyst Antigens and Their Validity in the Detection of Immunogenic Patterns of Cyclosporiasis Patients

Abstract

Introduction

The diagnosis of cyclosporiasis is currently based on the microscopic detection of oocysts, which may provide invalid results. The availability of simple, objective immunological screening tests would facilitate epidemiological studies of cyclosporiasis. Therefore, the present study aimed to identify the antigens of Cyclospora cayetanensis oocysts and their validity in serodiagnosis.

Methods

According to parasitological and molecular diagnoses, three study groups were specified. Group (G) I included 30 patients with cyclosporiasis, GII included 12 patients with other parasitic infections, and GIII included 16 healthy subjects. SDS-PAGE was used to analyse C. cayetanensis antigens, and the validity of western blotting and enzyme-linked immunosorbent assays (ELISAs) was then assessed amongst the sera of all study groups.

Results

The C. cayetanensis antigenic profile showed eight characteristic bands with molecular weights ranging from 14 to 175 kDa. Western blot analysis of sera revealed 93.3% (28/30 of GI) and 92.8% (26/28 of GII and III) sensitivity and specificity, respectively, dividing the patients in GI into four subgroups. The most frequent diagnostic bands (71.4% of GI sera) showed weights of 26–28 kDa, followed by 71 kDa (53.6%). ELISA sensitivity was 90% (27/30), and specificity was 78.6%. Validation showed perfect agreement between the PCR and western blot results, and ELISA presented substantial agreement with both the PCR and western blot results.

Conclusions

Our findings suggest the existence of high immunogenic diversity in C. cayetanensis and indicate that the 26–28 kDa immunogenic groups may potentially be used as a diagnostic marker of cyclosporiasis. Due to the high validity of ELISA, it might be the test of choice for the routine serodiagnosis of cyclosporiasis.

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Acknowledgements

We would like to convey our great thanks to Prof. Omnia Kandel and Dr. Mohammed Abd Elmoghny, Veterinary Section, National Research Centre, Egypt, for their critical and kind help in the practical work (western blotting and ELISA) of this study.

Funding

This research did not receive any specific grants from funding agencies in the public, commercial, or not-for-profit sectors.

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Authors

Contributions

Eman M Hussein contributed to the study conception and design. Maha M Al-Abbassy, Amira B Mokhtar, Ola A. Ismail and Eman M Hussein performed material preparation and data collection and analysis. Maha M Al-Abbassy and Eman M Hussein wrote the first draft of the manuscript. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Eman M. Hussein.

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The authors declare that they have no conflict of interest.

Ethical Approval

The patients and the parents of the neonates gave written consent before the beginning of sample collection. All procedures were carried out according to the ethical standards approved by the Institutional Human Ethics Committee, Faculty of Medicine, Suez Canal University, Egypt. All procedures related to animals were performed according to the ethical standards approved by the Institutional Animals Ethics Committee of the National Research Centre, Egypt, and the Faculty of Medicine, Suez Canal University, Egypt. The procedures used in this study adhered to the tenets of the Declaration of Helsinki.

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Hussein, E.M., El-Gayar, E.K., Ismail, O.A. et al. Identification of a Cyclospora cayetanensis Oocyst Antigens and Their Validity in the Detection of Immunogenic Patterns of Cyclosporiasis Patients. Acta Parasit. (2020). https://doi.org/10.1007/s11686-020-00289-w

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Keywords

  • C. cayetanensis
  • Oocysts
  • Antigens
  • SDS-PAGE
  • Western blotting
  • ELISA