Advancing syphilis diagnosis: multi-phase study evaluation of a TpN17-based double-antigen sandwich ELISA for detecting Treponema pallidum specific antibodies
Syphilis, a sexually transmitted infection caused by the bacterium Treponema pallidum, has high incidence rates among adults, pregnant women, and newborns. Diagnostic procedures typically involve a treponemal test (such as ELISA, CMIA, and IFI), followed by a non-treponemal test (VDRL and RPR). This...
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Frontiers Media S.A.
2025-04-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1572785/full |
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| author | Ângelo Antônio Oliveira Silva Ângelo Antônio Oliveira Silva Ângelo Antônio Oliveira Silva Larissa Carvalho Medrado Vasconcelos Larissa Carvalho Medrado Vasconcelos Natália Erdens Maron Freitas Natália Erdens Maron Freitas Talita Andrade Oliva Miralba Freire Carvalho Ribeiro Silva Isadora Cristina Siqueira Isadora Cristina Siqueira Isadora Cristina Siqueira Edimilson Domingos Silva Keila Gisele Azevedo Figueiredo Santos Maria Amélia Virgens Lima Nilson Ivo Tonin Zanchin Nilson Ivo Tonin Zanchin Nilson Ivo Tonin Zanchin Fred Luciano Neves Santos Fred Luciano Neves Santos Fred Luciano Neves Santos |
| author_facet | Ângelo Antônio Oliveira Silva Ângelo Antônio Oliveira Silva Ângelo Antônio Oliveira Silva Larissa Carvalho Medrado Vasconcelos Larissa Carvalho Medrado Vasconcelos Natália Erdens Maron Freitas Natália Erdens Maron Freitas Talita Andrade Oliva Miralba Freire Carvalho Ribeiro Silva Isadora Cristina Siqueira Isadora Cristina Siqueira Isadora Cristina Siqueira Edimilson Domingos Silva Keila Gisele Azevedo Figueiredo Santos Maria Amélia Virgens Lima Nilson Ivo Tonin Zanchin Nilson Ivo Tonin Zanchin Nilson Ivo Tonin Zanchin Fred Luciano Neves Santos Fred Luciano Neves Santos Fred Luciano Neves Santos |
| author_sort | Ângelo Antônio Oliveira Silva |
| collection | DOAJ |
| description | Syphilis, a sexually transmitted infection caused by the bacterium Treponema pallidum, has high incidence rates among adults, pregnant women, and newborns. Diagnostic procedures typically involve a treponemal test (such as ELISA, CMIA, and IFI), followed by a non-treponemal test (VDRL and RPR). This study aimed to assess the diagnostic performance of a double antigen sandwich ELISA (DAgS-ELISA) using the recombinant protein TpN17, analyzing serum samples from both infected and not infected with T. pallidum. A total of 712 samples were deemed eligible and recharacterized using VDRL, ELISA, and FTA-ABS, with 613 ultimately included in the evaluation: 180 T. pallidum-positive, 169 T. pallidum-negative, and 264 positive samples for other diseases. The assay was standardized using checkerboard titration and evaluated based on the area under the ROC curve (AUC), sensitivity, specificity, accuracy, likelihood values, diagnostic ratio, and Cohen’s Kappa index (κ). In phase I, positive and negative samples showed statistical differences (p < 0.0001) for the TpN17 protein. The ROC curve (AUC) was 98.7% and Cohen’s Kappa of 0.91, indicating almost perfect agreement with the reference tests. Phase II results demonstrated an AUC of 97.5%, specificity of 100%, sensitivity of 88.9%, accuracy of 94.3%, a positive likelihood ratio of 1.512, a negative likelihood ratio of 0.11, and a diagnostic odds ratio of 13,600, with a Cohen’s Kappa of 0.89. Cross-reactivity was observed in samples positive for Chagas disease (11.5%), HBV (2.6%), HCV (6.4%), and HTLV-1/2 (6.8%). Overall, TpN17 exhibited high diagnostic performance across all clinical stages of syphilis. Future research should expand the sample panel and explore new proteins to enhance DAgS-ELISA’s effectiveness and applicability for syphilis diagnosis across diverse clinical settings. |
| format | Article |
| id | doaj-art-18a58d65fb344fbbadb3453a4dd8dd19 |
| institution | OA Journals |
| issn | 1664-302X |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj-art-18a58d65fb344fbbadb3453a4dd8dd192025-08-20T02:08:27ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-04-011610.3389/fmicb.2025.15727851572785Advancing syphilis diagnosis: multi-phase study evaluation of a TpN17-based double-antigen sandwich ELISA for detecting Treponema pallidum specific antibodiesÂngelo Antônio Oliveira Silva0Ângelo Antônio Oliveira Silva1Ângelo Antônio Oliveira Silva2Larissa Carvalho Medrado Vasconcelos3Larissa Carvalho Medrado Vasconcelos4Natália Erdens Maron Freitas5Natália Erdens Maron Freitas6Talita Andrade Oliva7Miralba Freire Carvalho Ribeiro Silva8Isadora Cristina Siqueira9Isadora Cristina Siqueira10Isadora Cristina Siqueira11Edimilson Domingos Silva12Keila Gisele Azevedo Figueiredo Santos13Maria Amélia Virgens Lima14Nilson Ivo Tonin Zanchin15Nilson Ivo Tonin Zanchin16Nilson Ivo Tonin Zanchin17Fred Luciano Neves Santos18Fred Luciano Neves Santos19Fred Luciano Neves Santos20Advanced Public Health Laboratory, Gonçalo Moniz Institute, Oswaldo Cruz Foundation (Fiocruz-BA), Salvador, BrazilInterdisciplinary Research Group in Biotechnology and Epidemiology of Infectious Diseases (GRUPIBE), Gonçalo Moniz Institute, Oswaldo Cruz Foundation (FIOCRUZ-BA), Salvador, BrazilMedicine Course, Salvador University (UNIFACS), Salvador, BrazilAdvanced Public Health Laboratory, Gonçalo Moniz Institute, Oswaldo Cruz Foundation (Fiocruz-BA), Salvador, BrazilInterdisciplinary Research Group in Biotechnology and Epidemiology of Infectious Diseases (GRUPIBE), Gonçalo Moniz Institute, Oswaldo Cruz Foundation (FIOCRUZ-BA), Salvador, BrazilAdvanced Public Health Laboratory, Gonçalo Moniz Institute, Oswaldo Cruz Foundation (Fiocruz-BA), Salvador, BrazilInterdisciplinary Research Group in Biotechnology and Epidemiology of Infectious Diseases (GRUPIBE), Gonçalo Moniz Institute, Oswaldo Cruz Foundation (FIOCRUZ-BA), Salvador, BrazilState Center for Diagnosis, Assistance, and Research (CEDAP), Bahia State Health Department (SESAB), Salvador, BrazilState Center for Diagnosis, Assistance, and Research (CEDAP), Bahia State Health Department (SESAB), Salvador, BrazilInterdisciplinary Research Group in Biotechnology and Epidemiology of Infectious Diseases (GRUPIBE), Gonçalo Moniz Institute, Oswaldo Cruz Foundation (FIOCRUZ-BA), Salvador, BrazilLaboratory of Investigation in Global Health and Neglected Diseases, Gonçalo Moniz Institute, Oswaldo Cruz Foundation (FIOCRUZ-BA), Salvador, BrazilIntegrated Translational Program in Chagas Disease from Fiocruz (Fio-Chagas), Oswaldo Cruz Foundation (Fiocruz-RJ), Rio de Janeiro, BrazilDiagnostic Technology Laboratory, Immunobiological Technology Institute (Bio-Manguinhos), Oswaldo Cruz Foundation (Fiocruz-RJ), Rio de Janeiro, BrazilDiagnostic Technology Laboratory, Immunobiological Technology Institute (Bio-Manguinhos), Oswaldo Cruz Foundation (Fiocruz-RJ), Rio de Janeiro, BrazilDiagnostic Technology Laboratory, Immunobiological Technology Institute (Bio-Manguinhos), Oswaldo Cruz Foundation (Fiocruz-RJ), Rio de Janeiro, BrazilInterdisciplinary Research Group in Biotechnology and Epidemiology of Infectious Diseases (GRUPIBE), Gonçalo Moniz Institute, Oswaldo Cruz Foundation (FIOCRUZ-BA), Salvador, BrazilIntegrated Translational Program in Chagas Disease from Fiocruz (Fio-Chagas), Oswaldo Cruz Foundation (Fiocruz-RJ), Rio de Janeiro, BrazilStructural Biology and Protein Engineering Laboratory, Carlos Chagas Institute, Oswaldo Cruz Foundation (Fiocruz-PR), Curitiba, BrazilAdvanced Public Health Laboratory, Gonçalo Moniz Institute, Oswaldo Cruz Foundation (Fiocruz-BA), Salvador, BrazilInterdisciplinary Research Group in Biotechnology and Epidemiology of Infectious Diseases (GRUPIBE), Gonçalo Moniz Institute, Oswaldo Cruz Foundation (FIOCRUZ-BA), Salvador, BrazilIntegrated Translational Program in Chagas Disease from Fiocruz (Fio-Chagas), Oswaldo Cruz Foundation (Fiocruz-RJ), Rio de Janeiro, BrazilSyphilis, a sexually transmitted infection caused by the bacterium Treponema pallidum, has high incidence rates among adults, pregnant women, and newborns. Diagnostic procedures typically involve a treponemal test (such as ELISA, CMIA, and IFI), followed by a non-treponemal test (VDRL and RPR). This study aimed to assess the diagnostic performance of a double antigen sandwich ELISA (DAgS-ELISA) using the recombinant protein TpN17, analyzing serum samples from both infected and not infected with T. pallidum. A total of 712 samples were deemed eligible and recharacterized using VDRL, ELISA, and FTA-ABS, with 613 ultimately included in the evaluation: 180 T. pallidum-positive, 169 T. pallidum-negative, and 264 positive samples for other diseases. The assay was standardized using checkerboard titration and evaluated based on the area under the ROC curve (AUC), sensitivity, specificity, accuracy, likelihood values, diagnostic ratio, and Cohen’s Kappa index (κ). In phase I, positive and negative samples showed statistical differences (p < 0.0001) for the TpN17 protein. The ROC curve (AUC) was 98.7% and Cohen’s Kappa of 0.91, indicating almost perfect agreement with the reference tests. Phase II results demonstrated an AUC of 97.5%, specificity of 100%, sensitivity of 88.9%, accuracy of 94.3%, a positive likelihood ratio of 1.512, a negative likelihood ratio of 0.11, and a diagnostic odds ratio of 13,600, with a Cohen’s Kappa of 0.89. Cross-reactivity was observed in samples positive for Chagas disease (11.5%), HBV (2.6%), HCV (6.4%), and HTLV-1/2 (6.8%). Overall, TpN17 exhibited high diagnostic performance across all clinical stages of syphilis. Future research should expand the sample panel and explore new proteins to enhance DAgS-ELISA’s effectiveness and applicability for syphilis diagnosis across diverse clinical settings.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1572785/fullsyphilistreponemal testrecombinant proteinTpN17DAgS-ELISA |
| spellingShingle | Ângelo Antônio Oliveira Silva Ângelo Antônio Oliveira Silva Ângelo Antônio Oliveira Silva Larissa Carvalho Medrado Vasconcelos Larissa Carvalho Medrado Vasconcelos Natália Erdens Maron Freitas Natália Erdens Maron Freitas Talita Andrade Oliva Miralba Freire Carvalho Ribeiro Silva Isadora Cristina Siqueira Isadora Cristina Siqueira Isadora Cristina Siqueira Edimilson Domingos Silva Keila Gisele Azevedo Figueiredo Santos Maria Amélia Virgens Lima Nilson Ivo Tonin Zanchin Nilson Ivo Tonin Zanchin Nilson Ivo Tonin Zanchin Fred Luciano Neves Santos Fred Luciano Neves Santos Fred Luciano Neves Santos Advancing syphilis diagnosis: multi-phase study evaluation of a TpN17-based double-antigen sandwich ELISA for detecting Treponema pallidum specific antibodies Frontiers in Microbiology syphilis treponemal test recombinant protein TpN17 DAgS-ELISA |
| title | Advancing syphilis diagnosis: multi-phase study evaluation of a TpN17-based double-antigen sandwich ELISA for detecting Treponema pallidum specific antibodies |
| title_full | Advancing syphilis diagnosis: multi-phase study evaluation of a TpN17-based double-antigen sandwich ELISA for detecting Treponema pallidum specific antibodies |
| title_fullStr | Advancing syphilis diagnosis: multi-phase study evaluation of a TpN17-based double-antigen sandwich ELISA for detecting Treponema pallidum specific antibodies |
| title_full_unstemmed | Advancing syphilis diagnosis: multi-phase study evaluation of a TpN17-based double-antigen sandwich ELISA for detecting Treponema pallidum specific antibodies |
| title_short | Advancing syphilis diagnosis: multi-phase study evaluation of a TpN17-based double-antigen sandwich ELISA for detecting Treponema pallidum specific antibodies |
| title_sort | advancing syphilis diagnosis multi phase study evaluation of a tpn17 based double antigen sandwich elisa for detecting treponema pallidum specific antibodies |
| topic | syphilis treponemal test recombinant protein TpN17 DAgS-ELISA |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1572785/full |
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