Identification of putative new Escherichia coli flagellar antigens from human origin using serology, PCR-RFLP and DNA sequencing methods
Escherichia coli has been isolated frequently, showing flagellar antigens that are not recognized by any of the 53 antisera, provided by the most important reference center of E. coli, The International Escherichia and Klebsiella Center (WHO) of the Statens Serum Institute, Copenhagen, Denmark. The...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2011-03-01
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| Series: | Brazilian Journal of Infectious Diseases |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S141386701170160X |
| _version_ | 1811204418986573824 |
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| author | Monique Ribeiro Tiba Claúdia de Moura Marcelo Falsarella Carazzolle Domingos da Silva Leite |
| author_facet | Monique Ribeiro Tiba Claúdia de Moura Marcelo Falsarella Carazzolle Domingos da Silva Leite |
| author_sort | Monique Ribeiro Tiba |
| collection | DOAJ |
| description | Escherichia coli has been isolated frequently, showing flagellar antigens that are not recognized by any of the 53 antisera, provided by the most important reference center of E. coli, The International Escherichia and Klebsiella Center (WHO) of the Statens Serum Institute, Copenhagen, Denmark. The objective of this study was to characterize flagellar antigens of E. coli that express non-typeable H antigens. The methods used were serology, PCR-RFLP and DNA sequencing. This characterization was performed by gene amplification of the fliC (flagellin protein) by polymerase chain reaction in all 53 standards E.coli strains for the H antigens and 20 E. coli strains for which the H antigen was untypeable. The amplicons were digested by restriction enzymes, and different restriction enzyme profiles were observed. Anti-sera were produced in rabbits, for the non-typeable strains, and agglutination tests were carried out. In conclusion,the results showed that although non-typeable and typable H antigens strains had similar flagellar antigens, the two types of strains were distinct in terms of nucleotide sequence, and did not phenotypically react with the standard antiserum, as expected. Thirteen strains had been characterized as likely putative new H antigen using PCR-RFLP techniques, DNA sequencing and/or serology. Keywords: Escherichia coli, antigens, bacterial, polymerase chain reaction, polymorphism, restriction fragment length |
| first_indexed | 2024-04-12T03:12:50Z |
| format | Article |
| id | doaj.art-1df8813cd1c04430aefcad6cf5c29ddd |
| institution | Directory Open Access Journal |
| issn | 1413-8670 |
| language | English |
| last_indexed | 2024-04-12T03:12:50Z |
| publishDate | 2011-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Brazilian Journal of Infectious Diseases |
| spelling | doaj.art-1df8813cd1c04430aefcad6cf5c29ddd2022-12-22T03:50:16ZengElsevierBrazilian Journal of Infectious Diseases1413-86702011-03-01152144150Identification of putative new Escherichia coli flagellar antigens from human origin using serology, PCR-RFLP and DNA sequencing methodsMonique Ribeiro Tiba0Claúdia de Moura1Marcelo Falsarella Carazzolle2Domingos da Silva Leite3MSc; Dr.; Post-doctorate, Universidade Estadual de Campinas - UNICAMP, São Paulo, Brazil; Correspondence to: Monique Ribeiro Tiba Rua Visconde de Taunay, 147/41, Vila Itapura, Campinas, SP, Brazil.MSc; PhD Candidate, UNICAMP, São Paulo, BrazilMSc, Dr.; Physicist, UNICAMP, São Paulo, BrazilMSc, Dr.; Professor, UNICAMP, São Paulo, BrazilEscherichia coli has been isolated frequently, showing flagellar antigens that are not recognized by any of the 53 antisera, provided by the most important reference center of E. coli, The International Escherichia and Klebsiella Center (WHO) of the Statens Serum Institute, Copenhagen, Denmark. The objective of this study was to characterize flagellar antigens of E. coli that express non-typeable H antigens. The methods used were serology, PCR-RFLP and DNA sequencing. This characterization was performed by gene amplification of the fliC (flagellin protein) by polymerase chain reaction in all 53 standards E.coli strains for the H antigens and 20 E. coli strains for which the H antigen was untypeable. The amplicons were digested by restriction enzymes, and different restriction enzyme profiles were observed. Anti-sera were produced in rabbits, for the non-typeable strains, and agglutination tests were carried out. In conclusion,the results showed that although non-typeable and typable H antigens strains had similar flagellar antigens, the two types of strains were distinct in terms of nucleotide sequence, and did not phenotypically react with the standard antiserum, as expected. Thirteen strains had been characterized as likely putative new H antigen using PCR-RFLP techniques, DNA sequencing and/or serology. Keywords: Escherichia coli, antigens, bacterial, polymerase chain reaction, polymorphism, restriction fragment lengthhttp://www.sciencedirect.com/science/article/pii/S141386701170160X |
| spellingShingle | Monique Ribeiro Tiba Claúdia de Moura Marcelo Falsarella Carazzolle Domingos da Silva Leite Identification of putative new Escherichia coli flagellar antigens from human origin using serology, PCR-RFLP and DNA sequencing methods Brazilian Journal of Infectious Diseases |
| title | Identification of putative new Escherichia coli flagellar antigens from human origin using serology, PCR-RFLP and DNA sequencing methods |
| title_full | Identification of putative new Escherichia coli flagellar antigens from human origin using serology, PCR-RFLP and DNA sequencing methods |
| title_fullStr | Identification of putative new Escherichia coli flagellar antigens from human origin using serology, PCR-RFLP and DNA sequencing methods |
| title_full_unstemmed | Identification of putative new Escherichia coli flagellar antigens from human origin using serology, PCR-RFLP and DNA sequencing methods |
| title_short | Identification of putative new Escherichia coli flagellar antigens from human origin using serology, PCR-RFLP and DNA sequencing methods |
| title_sort | identification of putative new escherichia coli flagellar antigens from human origin using serology pcr rflp and dna sequencing methods |
| url | http://www.sciencedirect.com/science/article/pii/S141386701170160X |
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