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. 2017 Dec 29;17(1):807.
doi: 10.1186/s12879-017-2920-9.

An evaluation of a recombinant multiepitope based antigen for detection of Toxoplasma gondii specific antibodies

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An evaluation of a recombinant multiepitope based antigen for detection of Toxoplasma gondii specific antibodies

Khalid Hajissa et al. BMC Infect Dis. .

Abstract

Background: The inefficiency of the current tachyzoite antigen-based serological assays for the serodiagnosis of Toxoplasma gondii infection mandates the need for acquirement of reliable and standard diagnostic reagents. Recently, epitope-based antigens have emerged as an alternative diagnostic marker for the achievement of highly sensitive and specific capture antigens. In this study, the diagnostic utility of a recombinant multiepitope antigen (USM.TOXO1) for the serodiagnosis of human toxoplasmosis was evaluated.

Methods: An indirect enzyme-linked immunosorbent assay (ELISA) was developed to evaluate the usefulness of USM.TOXO1 antigen for the detection of IgG antibodies against Toxoplasma gondii in human sera. Whereas the reactivity of the developed antigen against IgM antibody was evaluated by western blot and Dot enzyme immunoassay (dot-EIA) analysis.

Results: The diagnostic performance of the new antigens in IgG ELISA was achieved at the maximum values of 85.43% and 81.25% for diagnostic sensitivity and specificity respectively. The USM.TOXO1 was also proven to be reactive with anti- T. gondii IgM antibody.

Conclusions: This finding makes the USM.TOXO1 antigen an attractive candidate for improving the toxoplasmosis serodiagnosis and demonstrates that multiepitope antigens could be a potential and promising diagnostic marker for the development of high sensitive and accurate assays.

Keywords: Elisa; Multiepitope; Recombinant antigens; Serodiagnosis; T. Gondii; USM.TOXO1.

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Conflict of interest statement

Ethics approval and consent to participate

The study protocol was approved by Human Research Ethics Committee of Universiti Sains Malaysia (HREC) (Approval number: USM/JEPeM/15020034). The consent was waived by the ethics committee since this study was carried out using the remaining of the sera requested for routine T.gondii serology detected by Elecsys® Toxoplasma IgG and IgM immunoassays as Gold Standard test. Similarly for the healthy control we used the remaining sera sent from blood donor for routine serological investigation such as VDRL for syphilis.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Construction of the USM.TOXO1 synthetic gene by assembly PCR. Lane M: 100 bp DNA Marker. Lane 1: 1st PCR product. Lane 2: Expected USM.TOXO1 gene (456 bp) amplified in the 2nd PCR. Lane N: Negative control
Fig. 2
Fig. 2
Reactivity of USM.TOXO1 with T. gondii IgM antibody by using immunoblots. a Western blot: Lane M: Molecular weight marker, Strips 1–3: Results of 3 positive sera, Strips 4–5: Results of negative sera b Dot-EIA: Strips 1–5: Results of positive sera, Strips 6–10: Results of negative sera

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