Development of a new serotyping ELISA for Toxoplasma gondii type II, type III and Africa 1 lineages using in silico peptide discovery methods, well categorized feline and human outbreak serum samples
Abstract Background Discovery of new Toxoplasma gondii serotyping epitopes is important due to reports showing the influence of genotype on the severity of toxoplasmosis. In Turkey, genotypes belonging to type II, type III and Africa 1 lineages were mainly detected. The present study focused on to f...
| Main Authors: | , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-01-01
|
| Series: | BMC Infectious Diseases |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12879-022-07088-w |
| _version_ | 1798022610898911232 |
|---|---|
| author | Hüseyin Can Ayşegül Aksoy Gökmen Mert Döşkaya Sedef Erkunt Alak Aysu Değirmenci Döşkaya Muhammet Karakavuk Ahmet Efe Köseoğlu Tuğba Karakavuk Ceren Gül Mervenur Güvendi Aytül Gül Adnan Yüksel Gürüz Selçuk Kaya Aurélien Mercier Cemal Ün |
| author_facet | Hüseyin Can Ayşegül Aksoy Gökmen Mert Döşkaya Sedef Erkunt Alak Aysu Değirmenci Döşkaya Muhammet Karakavuk Ahmet Efe Köseoğlu Tuğba Karakavuk Ceren Gül Mervenur Güvendi Aytül Gül Adnan Yüksel Gürüz Selçuk Kaya Aurélien Mercier Cemal Ün |
| author_sort | Hüseyin Can |
| collection | DOAJ |
| description | Abstract Background Discovery of new Toxoplasma gondii serotyping epitopes is important due to reports showing the influence of genotype on the severity of toxoplasmosis. In Turkey, genotypes belonging to type II, type III and Africa 1 lineages were mainly detected. The present study focused on to find out epitopes with high discriminative capacity to serotype these genotypes using well characterized strains isolated from Turkey. Methods To meet this objective, GRA6 and GRA7 genes were sequenced from strains belonging to the type II, III and Africa 1 lineages, and B cell epitopes inside these sequences were predicted by Bcepred and additional docking analysis was performed with B cell receptor. Based on these analyses, 22 peptides harboring lineage specific epitopes were synthesized. Then, the serotyping potency of these peptides was tested using peptide ELISA and well categorized serum samples collected from stray cats infected with genotypes of the different lineages type II (n:9), III (n:1) and Africa 1 (n:1). As a result of peptide-ELISA, a serotyping schema was constructed with peptides that show high discriminative capacity and this assay was validated by sera collected from humans after an outbreak (n:30) and mother/newborn pair sera (n:3). Later, the validated serotyping schema was used to serotype a larger group of human (n:38) and cat (n:24) sera. Results Among 22 peptides, GRA6II/c, GRA7III/d, and GRA6 Africa 1/b epitopes have shown discriminative capacity. During the validation of peptide-ELISA, the serotype of toxoplasmosis outbreak and mother/newborn cases were detected to be serotype II. Moreover, the analyses in a larger group showed that serotype II was prevalent in humans and stray cats. Conclusions Overall, the results showed that the serotyping schema could be successfully used to serotype T. gondii infections caused by type II, III and Africa 1 genotype. |
| first_indexed | 2024-04-11T17:32:45Z |
| format | Article |
| id | doaj.art-da7c5e8a2d2040bcb016417475638fea |
| institution | Directory Open Access Journal |
| issn | 1471-2334 |
| language | English |
| last_indexed | 2024-04-11T17:32:45Z |
| publishDate | 2022-01-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Infectious Diseases |
| spelling | doaj.art-da7c5e8a2d2040bcb016417475638fea2022-12-22T04:11:58ZengBMCBMC Infectious Diseases1471-23342022-01-0122111410.1186/s12879-022-07088-wDevelopment of a new serotyping ELISA for Toxoplasma gondii type II, type III and Africa 1 lineages using in silico peptide discovery methods, well categorized feline and human outbreak serum samplesHüseyin Can0Ayşegül Aksoy Gökmen1Mert Döşkaya2Sedef Erkunt Alak3Aysu Değirmenci Döşkaya4Muhammet Karakavuk5Ahmet Efe Köseoğlu6Tuğba Karakavuk7Ceren Gül8Mervenur Güvendi9Aytül Gül10Adnan Yüksel Gürüz11Selçuk Kaya12Aurélien Mercier13Cemal Ün14Molecular Biology Section, Department of Biology, Faculty of Science, Ege UniversityDepartment of Microbiology, Faculty of Medicine, İzmir Katip Çelebi UniversityVaccine Development, Application and Research Center, Ege UniversityMolecular Biology Section, Department of Biology, Faculty of Science, Ege UniversityVaccine Development, Application and Research Center, Ege UniversityVaccine Development, Application and Research Center, Ege UniversityMolecular Biology Section, Department of Biology, Faculty of Science, Ege UniversityGraduate Faculty of Natural and Applied Science Biotechnology Program, Ege UniversityGraduate Faculty of Natural and Applied Science Biotechnology Program, Ege UniversityMolecular Biology Section, Department of Biology, Faculty of Science, Ege UniversityDepartment of Bioengineering, Faculty of Engineering, Ege UniversityVaccine Development, Application and Research Center, Ege UniversityDepartment of Microbiology, Faculty of Medicine, İzmir Katip Çelebi UniversityCentre National de Référence (CNR) Toxoplasmose/Toxoplasma Biological Resource Center (BRC), Centre Hospitalier-Universitaire DupuytrenMolecular Biology Section, Department of Biology, Faculty of Science, Ege UniversityAbstract Background Discovery of new Toxoplasma gondii serotyping epitopes is important due to reports showing the influence of genotype on the severity of toxoplasmosis. In Turkey, genotypes belonging to type II, type III and Africa 1 lineages were mainly detected. The present study focused on to find out epitopes with high discriminative capacity to serotype these genotypes using well characterized strains isolated from Turkey. Methods To meet this objective, GRA6 and GRA7 genes were sequenced from strains belonging to the type II, III and Africa 1 lineages, and B cell epitopes inside these sequences were predicted by Bcepred and additional docking analysis was performed with B cell receptor. Based on these analyses, 22 peptides harboring lineage specific epitopes were synthesized. Then, the serotyping potency of these peptides was tested using peptide ELISA and well categorized serum samples collected from stray cats infected with genotypes of the different lineages type II (n:9), III (n:1) and Africa 1 (n:1). As a result of peptide-ELISA, a serotyping schema was constructed with peptides that show high discriminative capacity and this assay was validated by sera collected from humans after an outbreak (n:30) and mother/newborn pair sera (n:3). Later, the validated serotyping schema was used to serotype a larger group of human (n:38) and cat (n:24) sera. Results Among 22 peptides, GRA6II/c, GRA7III/d, and GRA6 Africa 1/b epitopes have shown discriminative capacity. During the validation of peptide-ELISA, the serotype of toxoplasmosis outbreak and mother/newborn cases were detected to be serotype II. Moreover, the analyses in a larger group showed that serotype II was prevalent in humans and stray cats. Conclusions Overall, the results showed that the serotyping schema could be successfully used to serotype T. gondii infections caused by type II, III and Africa 1 genotype.https://doi.org/10.1186/s12879-022-07088-wToxoplasmosisSerotypingGenotypingGRA6GRA7Peptide ELISA |
| spellingShingle | Hüseyin Can Ayşegül Aksoy Gökmen Mert Döşkaya Sedef Erkunt Alak Aysu Değirmenci Döşkaya Muhammet Karakavuk Ahmet Efe Köseoğlu Tuğba Karakavuk Ceren Gül Mervenur Güvendi Aytül Gül Adnan Yüksel Gürüz Selçuk Kaya Aurélien Mercier Cemal Ün Development of a new serotyping ELISA for Toxoplasma gondii type II, type III and Africa 1 lineages using in silico peptide discovery methods, well categorized feline and human outbreak serum samples BMC Infectious Diseases Toxoplasmosis Serotyping Genotyping GRA6 GRA7 Peptide ELISA |
| title | Development of a new serotyping ELISA for Toxoplasma gondii type II, type III and Africa 1 lineages using in silico peptide discovery methods, well categorized feline and human outbreak serum samples |
| title_full | Development of a new serotyping ELISA for Toxoplasma gondii type II, type III and Africa 1 lineages using in silico peptide discovery methods, well categorized feline and human outbreak serum samples |
| title_fullStr | Development of a new serotyping ELISA for Toxoplasma gondii type II, type III and Africa 1 lineages using in silico peptide discovery methods, well categorized feline and human outbreak serum samples |
| title_full_unstemmed | Development of a new serotyping ELISA for Toxoplasma gondii type II, type III and Africa 1 lineages using in silico peptide discovery methods, well categorized feline and human outbreak serum samples |
| title_short | Development of a new serotyping ELISA for Toxoplasma gondii type II, type III and Africa 1 lineages using in silico peptide discovery methods, well categorized feline and human outbreak serum samples |
| title_sort | development of a new serotyping elisa for toxoplasma gondii type ii type iii and africa 1 lineages using in silico peptide discovery methods well categorized feline and human outbreak serum samples |
| topic | Toxoplasmosis Serotyping Genotyping GRA6 GRA7 Peptide ELISA |
| url | https://doi.org/10.1186/s12879-022-07088-w |
| work_keys_str_mv | AT huseyincan developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT aysegulaksoygokmen developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT mertdoskaya developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT sedeferkuntalak developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT aysudegirmencidoskaya developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT muhammetkarakavuk developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT ahmetefekoseoglu developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT tugbakarakavuk developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT cerengul developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT mervenurguvendi developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT aytulgul developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT adnanyukselguruz developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT selcukkaya developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT aurelienmercier developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples AT cemalun developmentofanewserotypingelisafortoxoplasmagondiitypeiitypeiiiandafrica1lineagesusinginsilicopeptidediscoverymethodswellcategorizedfelineandhumanoutbreakserumsamples |