Cargando…
A Search for Trypanosoma brucei rhodesiense Diagnostic Antigens by Proteomic Screening and Targeted Cloning
BACKGROUND: The only available diagnostic method for East African trypanosomiasis is light microscopy of blood samples. A simple immunodiagnostic would greatly aid trypanosomiasis control. METHODOLOGY AND PRINCIPAL FINDINGS: To find trypanosome proteins that are specifically recognised by sera from...
Autores principales: | , , , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2010
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2835760/ https://www.ncbi.nlm.nih.gov/pubmed/20224787 http://dx.doi.org/10.1371/journal.pone.0009630 |
_version_ | 1782178664909111296 |
---|---|
author | Manful, Theresa Mulindwa, Julius Frank, Fernanda M. Clayton, Christine E. Matovu, Enock |
author_facet | Manful, Theresa Mulindwa, Julius Frank, Fernanda M. Clayton, Christine E. Matovu, Enock |
author_sort | Manful, Theresa |
collection | PubMed |
description | BACKGROUND: The only available diagnostic method for East African trypanosomiasis is light microscopy of blood samples. A simple immunodiagnostic would greatly aid trypanosomiasis control. METHODOLOGY AND PRINCIPAL FINDINGS: To find trypanosome proteins that are specifically recognised by sera from human sleeping sickness patients, we have screened the Trypanosoma brucei brucei proteome by Western blotting. Using cytosolic, cytoskeletal and glycosomal fractions, we found that the vast majority of abundant trypanosome proteins is not specifically recognised by patient sera. We identified phosphoglycerate kinase (PGKC), heat shock protein (HSP70), and histones H2B and H3 as possible candidate diagnostic antigens. These proteins, plus paraflagellar rod protein 1, rhodesain (a cysteine protease), and an extracellular fragment of the Trypanosoma brucei nucleoside transporter TbNT10, were expressed in E. coli and tested for reactivity with patient and control sera. Only TbHSP70 was preferentially recognized by patient sera, but the sensitivity and specificity were insufficient for use of TbHSP70 alone as a diagnostic. Immunoprecipitation using a native protein extract revealed no specifically reacting proteins. CONCLUSIONS: No abundant T. brucei soluble, glycosomal or cytoskeletal protein is likely to be useful in diagnosis. To find useful diagnostic antigens it will therefore be necessary to use more sophisticated proteomic methods, or to test a very large panel of candidate proteins. |
format | Text |
id | pubmed-2835760 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-28357602010-03-12 A Search for Trypanosoma brucei rhodesiense Diagnostic Antigens by Proteomic Screening and Targeted Cloning Manful, Theresa Mulindwa, Julius Frank, Fernanda M. Clayton, Christine E. Matovu, Enock PLoS One Research Article BACKGROUND: The only available diagnostic method for East African trypanosomiasis is light microscopy of blood samples. A simple immunodiagnostic would greatly aid trypanosomiasis control. METHODOLOGY AND PRINCIPAL FINDINGS: To find trypanosome proteins that are specifically recognised by sera from human sleeping sickness patients, we have screened the Trypanosoma brucei brucei proteome by Western blotting. Using cytosolic, cytoskeletal and glycosomal fractions, we found that the vast majority of abundant trypanosome proteins is not specifically recognised by patient sera. We identified phosphoglycerate kinase (PGKC), heat shock protein (HSP70), and histones H2B and H3 as possible candidate diagnostic antigens. These proteins, plus paraflagellar rod protein 1, rhodesain (a cysteine protease), and an extracellular fragment of the Trypanosoma brucei nucleoside transporter TbNT10, were expressed in E. coli and tested for reactivity with patient and control sera. Only TbHSP70 was preferentially recognized by patient sera, but the sensitivity and specificity were insufficient for use of TbHSP70 alone as a diagnostic. Immunoprecipitation using a native protein extract revealed no specifically reacting proteins. CONCLUSIONS: No abundant T. brucei soluble, glycosomal or cytoskeletal protein is likely to be useful in diagnosis. To find useful diagnostic antigens it will therefore be necessary to use more sophisticated proteomic methods, or to test a very large panel of candidate proteins. Public Library of Science 2010-03-10 /pmc/articles/PMC2835760/ /pubmed/20224787 http://dx.doi.org/10.1371/journal.pone.0009630 Text en Manful et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Manful, Theresa Mulindwa, Julius Frank, Fernanda M. Clayton, Christine E. Matovu, Enock A Search for Trypanosoma brucei rhodesiense Diagnostic Antigens by Proteomic Screening and Targeted Cloning |
title | A Search for Trypanosoma brucei rhodesiense Diagnostic Antigens by Proteomic Screening and Targeted Cloning |
title_full | A Search for Trypanosoma brucei rhodesiense Diagnostic Antigens by Proteomic Screening and Targeted Cloning |
title_fullStr | A Search for Trypanosoma brucei rhodesiense Diagnostic Antigens by Proteomic Screening and Targeted Cloning |
title_full_unstemmed | A Search for Trypanosoma brucei rhodesiense Diagnostic Antigens by Proteomic Screening and Targeted Cloning |
title_short | A Search for Trypanosoma brucei rhodesiense Diagnostic Antigens by Proteomic Screening and Targeted Cloning |
title_sort | search for trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2835760/ https://www.ncbi.nlm.nih.gov/pubmed/20224787 http://dx.doi.org/10.1371/journal.pone.0009630 |
work_keys_str_mv | AT manfultheresa asearchfortrypanosomabruceirhodesiensediagnosticantigensbyproteomicscreeningandtargetedcloning AT mulindwajulius asearchfortrypanosomabruceirhodesiensediagnosticantigensbyproteomicscreeningandtargetedcloning AT frankfernandam asearchfortrypanosomabruceirhodesiensediagnosticantigensbyproteomicscreeningandtargetedcloning AT claytonchristinee asearchfortrypanosomabruceirhodesiensediagnosticantigensbyproteomicscreeningandtargetedcloning AT matovuenock asearchfortrypanosomabruceirhodesiensediagnosticantigensbyproteomicscreeningandtargetedcloning AT manfultheresa searchfortrypanosomabruceirhodesiensediagnosticantigensbyproteomicscreeningandtargetedcloning AT mulindwajulius searchfortrypanosomabruceirhodesiensediagnosticantigensbyproteomicscreeningandtargetedcloning AT frankfernandam searchfortrypanosomabruceirhodesiensediagnosticantigensbyproteomicscreeningandtargetedcloning AT claytonchristinee searchfortrypanosomabruceirhodesiensediagnosticantigensbyproteomicscreeningandtargetedcloning AT matovuenock searchfortrypanosomabruceirhodesiensediagnosticantigensbyproteomicscreeningandtargetedcloning |