Design and expression of a chimeric recombinant antigen (SsIR-Ss1a) for the serodiagnosis of human strongyloidiasis: Evaluation of performance, sensitivity, and specificity.
<h4>Background</h4>The sensitivity of parasitological and molecular methods is unsatisfactory for the diagnosis of strongyloidiasis, and serological techniques are remaining as the most effective diagnostic approach. The present study aimed to design and produce a chimeric recombinant an...
Saved in:
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2024-07-01
|
| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | https://doi.org/10.1371/journal.pntd.0012320 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832540007551729664 |
|---|---|
| author | Mostafa Omidian Zohreh Mostafavi-Pour Marzieh Asadi Meysam Sharifdini Navid Nezafat Ali Pouryousef Amir Savardashtaki Mortaza Taheri-Anganeh Fattaneh Mikaeili Bahador Sarkari |
| author_facet | Mostafa Omidian Zohreh Mostafavi-Pour Marzieh Asadi Meysam Sharifdini Navid Nezafat Ali Pouryousef Amir Savardashtaki Mortaza Taheri-Anganeh Fattaneh Mikaeili Bahador Sarkari |
| author_sort | Mostafa Omidian |
| collection | DOAJ |
| description | <h4>Background</h4>The sensitivity of parasitological and molecular methods is unsatisfactory for the diagnosis of strongyloidiasis, and serological techniques are remaining as the most effective diagnostic approach. The present study aimed to design and produce a chimeric recombinant antigen from Strongyloides stercoralis immunoreactive antigen (SsIR) and Ss1a antigens, using immune-informatics approaches, and evaluated its diagnostic performance in an ELISA system for the diagnosis of human strongyloidiasis.<h4>Methodology/principal findings</h4>The coding sequences for SsIR and Ss1a were selected from GenBank and were gene-optimized. Using bioinformatics analysis, the regions with the highest antigenicity that did not overlap with other parasite antigens were selected. The chimeric recombinant antigen SsIR- Ss1a, was constructed. The solubility and physicochemical properties of the designed construct were analyzed and its tertiary structures were built and evaluated. The construct was expressed into the pET-23a (+) expression vector and the optimized DNA sequences of SsIR-Ss1a (873 bp) were cloned into competent E. coli DH5α cells. Diagnostic performances of the produced recombinant antigen, along with a commercial kit were evaluated in an indirect ELISA system, using a panel of sera from strongyloidiasis patients and controls. The physicochemical and bioinformatics evaluations revealed that the designed chimeric construct is soluble, has a molecular with of 35 KDa, and is antigenic. Western blotting confirmed the immunoreactivity of the produced chimeric recombinant antigen with the sera of strongyloidiasis patients. The sensitivity and specificity of the indirect ELISA system, using the produced SsIR-Ss1a chimeric antigen, were found to be 93.94% (95% CI, 0.803 to 0.989) and 97.22% (95% CI, 0.921 to 0.992) respectively.<h4>Conclusions/significance</h4>The preliminary findings of this study suggest that the produced SsIR-Ss1a chimeric antigen shows promise in the diagnosis of human strongyloidiasis. However, these results are based on a limited panel of samples, and further research with a larger sample size is necessary to confirm its accuracy. The construct has potential as an antigen in the ELISA system for the serological diagnosis of this neglected parasitic infection, but additional validation is required. |
| format | Article |
| id | doaj-art-e8e40258ad294a448bd319884821d75a |
| institution | Kabale University |
| issn | 1935-2727 1935-2735 |
| language | English |
| publishDate | 2024-07-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj-art-e8e40258ad294a448bd319884821d75a2025-02-05T05:33:29ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352024-07-01187e001232010.1371/journal.pntd.0012320Design and expression of a chimeric recombinant antigen (SsIR-Ss1a) for the serodiagnosis of human strongyloidiasis: Evaluation of performance, sensitivity, and specificity.Mostafa OmidianZohreh Mostafavi-PourMarzieh AsadiMeysam SharifdiniNavid NezafatAli PouryousefAmir SavardashtakiMortaza Taheri-AnganehFattaneh MikaeiliBahador Sarkari<h4>Background</h4>The sensitivity of parasitological and molecular methods is unsatisfactory for the diagnosis of strongyloidiasis, and serological techniques are remaining as the most effective diagnostic approach. The present study aimed to design and produce a chimeric recombinant antigen from Strongyloides stercoralis immunoreactive antigen (SsIR) and Ss1a antigens, using immune-informatics approaches, and evaluated its diagnostic performance in an ELISA system for the diagnosis of human strongyloidiasis.<h4>Methodology/principal findings</h4>The coding sequences for SsIR and Ss1a were selected from GenBank and were gene-optimized. Using bioinformatics analysis, the regions with the highest antigenicity that did not overlap with other parasite antigens were selected. The chimeric recombinant antigen SsIR- Ss1a, was constructed. The solubility and physicochemical properties of the designed construct were analyzed and its tertiary structures were built and evaluated. The construct was expressed into the pET-23a (+) expression vector and the optimized DNA sequences of SsIR-Ss1a (873 bp) were cloned into competent E. coli DH5α cells. Diagnostic performances of the produced recombinant antigen, along with a commercial kit were evaluated in an indirect ELISA system, using a panel of sera from strongyloidiasis patients and controls. The physicochemical and bioinformatics evaluations revealed that the designed chimeric construct is soluble, has a molecular with of 35 KDa, and is antigenic. Western blotting confirmed the immunoreactivity of the produced chimeric recombinant antigen with the sera of strongyloidiasis patients. The sensitivity and specificity of the indirect ELISA system, using the produced SsIR-Ss1a chimeric antigen, were found to be 93.94% (95% CI, 0.803 to 0.989) and 97.22% (95% CI, 0.921 to 0.992) respectively.<h4>Conclusions/significance</h4>The preliminary findings of this study suggest that the produced SsIR-Ss1a chimeric antigen shows promise in the diagnosis of human strongyloidiasis. However, these results are based on a limited panel of samples, and further research with a larger sample size is necessary to confirm its accuracy. The construct has potential as an antigen in the ELISA system for the serological diagnosis of this neglected parasitic infection, but additional validation is required.https://doi.org/10.1371/journal.pntd.0012320 |
| spellingShingle | Mostafa Omidian Zohreh Mostafavi-Pour Marzieh Asadi Meysam Sharifdini Navid Nezafat Ali Pouryousef Amir Savardashtaki Mortaza Taheri-Anganeh Fattaneh Mikaeili Bahador Sarkari Design and expression of a chimeric recombinant antigen (SsIR-Ss1a) for the serodiagnosis of human strongyloidiasis: Evaluation of performance, sensitivity, and specificity. PLoS Neglected Tropical Diseases |
| title | Design and expression of a chimeric recombinant antigen (SsIR-Ss1a) for the serodiagnosis of human strongyloidiasis: Evaluation of performance, sensitivity, and specificity. |
| title_full | Design and expression of a chimeric recombinant antigen (SsIR-Ss1a) for the serodiagnosis of human strongyloidiasis: Evaluation of performance, sensitivity, and specificity. |
| title_fullStr | Design and expression of a chimeric recombinant antigen (SsIR-Ss1a) for the serodiagnosis of human strongyloidiasis: Evaluation of performance, sensitivity, and specificity. |
| title_full_unstemmed | Design and expression of a chimeric recombinant antigen (SsIR-Ss1a) for the serodiagnosis of human strongyloidiasis: Evaluation of performance, sensitivity, and specificity. |
| title_short | Design and expression of a chimeric recombinant antigen (SsIR-Ss1a) for the serodiagnosis of human strongyloidiasis: Evaluation of performance, sensitivity, and specificity. |
| title_sort | design and expression of a chimeric recombinant antigen ssir ss1a for the serodiagnosis of human strongyloidiasis evaluation of performance sensitivity and specificity |
| url | https://doi.org/10.1371/journal.pntd.0012320 |
| work_keys_str_mv | AT mostafaomidian designandexpressionofachimericrecombinantantigenssirss1afortheserodiagnosisofhumanstrongyloidiasisevaluationofperformancesensitivityandspecificity AT zohrehmostafavipour designandexpressionofachimericrecombinantantigenssirss1afortheserodiagnosisofhumanstrongyloidiasisevaluationofperformancesensitivityandspecificity AT marziehasadi designandexpressionofachimericrecombinantantigenssirss1afortheserodiagnosisofhumanstrongyloidiasisevaluationofperformancesensitivityandspecificity AT meysamsharifdini designandexpressionofachimericrecombinantantigenssirss1afortheserodiagnosisofhumanstrongyloidiasisevaluationofperformancesensitivityandspecificity AT navidnezafat designandexpressionofachimericrecombinantantigenssirss1afortheserodiagnosisofhumanstrongyloidiasisevaluationofperformancesensitivityandspecificity AT alipouryousef designandexpressionofachimericrecombinantantigenssirss1afortheserodiagnosisofhumanstrongyloidiasisevaluationofperformancesensitivityandspecificity AT amirsavardashtaki designandexpressionofachimericrecombinantantigenssirss1afortheserodiagnosisofhumanstrongyloidiasisevaluationofperformancesensitivityandspecificity AT mortazataherianganeh designandexpressionofachimericrecombinantantigenssirss1afortheserodiagnosisofhumanstrongyloidiasisevaluationofperformancesensitivityandspecificity AT fattanehmikaeili designandexpressionofachimericrecombinantantigenssirss1afortheserodiagnosisofhumanstrongyloidiasisevaluationofperformancesensitivityandspecificity AT bahadorsarkari designandexpressionofachimericrecombinantantigenssirss1afortheserodiagnosisofhumanstrongyloidiasisevaluationofperformancesensitivityandspecificity |