Design and evaluation of a recombinant multi-epitope antigen for serodiagnosis of Toxoplasma gondii infection in humans

Particle and Fibre Toxicology

Zeehaida Mohamed²  Rapeah Suppian¹  Robaiza Zakaria²  Khalid Hajissa² 

[1] Biomedicine Program, School of Health Sciences, Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia;Department of Medical Microbiology & Parasitology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia

关键词: USM.TOXO1;    Toxoplasma gondii;    Synthetic gene;    Epitopes;    Assembly PCR;   

Others  :  1224206
DOI  :  10.1186/s13071-015-0932-0

 received in 2015-04-14, accepted in 2015-06-02,  发布年份 2015

【 摘 要 】

Background

Serological investigation remains the primary approach to achieve satisfactory results in Toxoplasma gondii identification. However, the accuracy of the native antigen used in the current diagnostic kits has proven to be insufficient as well as difficult to standardize, so significant efforts have been made to find alternative reagents as capture antigens. Consequently, multi-epitope peptides are promising diagnostic markers, with the potential for improving the accuracy of diagnostic kits. In this study, we described a simple, inexpensive and improved strategy to acquire such diagnostic markers. The study was aimed at producing novel synthetic protein consisting of multiple immunodominant epitopes of several T. gondii antigens.

Findings

To accomplish our goals, a single synthetic gene of approximately 456 bp, which encodes potential epitopes of T. gondii antigens, was successfully constructed using gene assembly PCR. The constructed gene was cloned into a pET32a expression vector and transformed into BL21 E. coli. The entire protein was successfully expressed and purified. Subsequently, the preliminary diagnostic performance of expressed protein was evaluated by developing IgG enzyme-linked immunosorbent assay (ELISA) and Western blot analysis using human sera. The results showed 100 % sensitivity and specificity.

Conclusion

A purified protein expressing multi-immunodominant epitopes of T. gondii was generated. Further studies are required to evaluate the immunogenicity in animal models and to verify the immuno-reactivity of USM.TOXO1 as a diagnostic antigen.

【 授权许可】

   
2015 Hajissa et al.

【 图 表 】

Fig. 1.

Fig. 2.

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