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Developing multi\u2010epitope antigens for serological assays offers the potential for enhancing diagnostic accuracy. This study aimed to design a novel multi\u2010epitope antigen for HCMV infection diagnosis using immunoinformatic approaches. Five tegument proteins (universal protein resource [UniProt] ID: Po8318, Po6725, F5HC97, Q6RX10, and F5HC05) were selected based on their antigenic properties and literature review. Six linear B\u2010cell epitopes were predicted within conserved regions of each antigen sequence and linked with appropriate linkers. The designed multi\u2010epitope antigen underwent thorough evaluation for physicochemical properties, solubility, antigenicity, and cross\u2010reactivity. Additionally, the three\u2010dimensional structure of the antigen was predicted, refined, and validated. The nucleotide sequence of the designed antigen was optimized for successful expression in <jats:italic>Escherichia coli<\/jats:italic> and inserted into a pET23a (+) vector. Immunoinformatic analysis revealed that the multi\u2010epitope antigen exhibits stability, antigenicity, and lacks cross\u2010reactivity. Our findings suggest that this multi\u2010epitope antigen is a promising candidate for diagnosing HCMV infection. However, further validation through laboratory testing is required to confirm its diagnostic efficacy.<\/jats:p>","DOI":"10.1002\/bab.2677","type":"journal-article","created":{"date-parts":[[2024,10,17]],"date-time":"2024-10-17T10:40:47Z","timestamp":1729161647000},"page":"469-483","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Designing a novel multi\u2010epitope antigen for diagnosing human cytomegalovirus infection: An immunoinformatics approach"],"prefix":"10.1002","volume":"72","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6140-1516","authenticated-orcid":false,"given":"Marzieh","family":"Asadi","sequence":"first","affiliation":[{"name":"Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies Shiraz University of Medical Sciences, Shiraz  Shiraz Iran"},{"name":"Student Research Committee Shiraz University of Medical Sciences  Shiraz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Younes","family":"Ghasemi","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Biotechnology, School of Pharmacy Shiraz University of Medical Sciences  Shiraz Iran"},{"name":"Pharmaceutical Sciences Research Center Shiraz University of Medical Science  Shiraz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8103-2685","authenticated-orcid":false,"given":"Navid","family":"Nezafat","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Biotechnology, School of Pharmacy Shiraz University of Medical Sciences  Shiraz Iran"},{"name":"Pharmaceutical Sciences Research Center Shiraz University of Medical Science  Shiraz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bahador","family":"Sarkari","sequence":"additional","affiliation":[{"name":"Department of Parasitology and Mycology, School of Medicine Shiraz University of Medical Science  Shiraz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maryam","family":"Baneshi","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies Shiraz University of Medical Sciences, Shiraz  Shiraz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3779-177X","authenticated-orcid":false,"given":"Zohreh","family":"Mostafavi\u2010Pour","sequence":"additional","affiliation":[{"name":"Recombinant Protein Laboratory, Department of Biochemistry, School of Medicine Shiraz University of Medical Sciences  Shiraz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammad Hossein","family":"Anbardar","sequence":"additional","affiliation":[{"name":"Department of Pathology, School of Medicine Shiraz University of Medical Sciences  Shiraz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1700-9426","authenticated-orcid":false,"given":"Amir","family":"Savardashtaki","sequence":"additional","affiliation":[{"name":"Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies Shiraz University of Medical Sciences, Shiraz  Shiraz Iran"},{"name":"Infertility Research Center Shiraz University of Medical Sciences  Shiraz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,10,17]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-021-00582-z"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms8050685"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajog.2020.02.018"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jiz601"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/bab.2308"},{"issue":"2","key":"e_1_2_11_7_1","first-page":"111","article-title":"Immunodiagnostic value of echinococcus granulosus recombinant B8\/1 subunit of antigen B","volume":"14","author":"Savardashtaki A","year":"2017","journal-title":"Iran J Immunol"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12934-024-02418-w"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2017.08.019"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcv.2017.10.004"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.2174\/1872208313666190716093911"},{"key":"e_1_2_11_12_1","volume-title":"BcePred: prediction of continuous B\u2010cell epitopes in antigenic sequences using physico\u2010chemical properties. 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