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Conference Paper: Development of DNA aptamers against Treponema denticola

TitleDevelopment of DNA aptamers against Treponema denticola
Authors
KeywordsDiagnosis
Microbiology
Molecular biology
Nucleic acids and Periodontal disease
Issue Date2014
PublisherSage Publications, Inc. The Journal's web site is located at http://www.sagepub.com/journalsProdDesc.nav?prodId=Journal201925
Citation
The 92nd General Session & Exhibition of the International Association for Dental Research (IADR), Cape Town, South Africa, 25-28 June 2014. In Journal of Dental Research, 2014, v. 93 n. Special issue B: abstract no. 388 How to Cite?
AbstractThe oral spirochete bacterium Treponema denticola is implicated in the etiology of periodontal disease. However, the elucidation of more specific disease correlations is hindered by the lack of reliable and discriminatory diagnostic methods for T. denticola cell detection and enumeration. Aptamers are an emerging class of molecular recognition agents, which have binding properties analogous to antibodies. Their potential application for oral microbial cell detection remains to be firmly established. Objective: To identify DNA aptamers against a cell-surface protein from Treponema denticola. Method: A recombinant form of an established cell surface protein from T. denticola ATCC 35405 was used as the target for the identification of single stranded DNA aptamers. A SELEX approach was used for aptamer selection from a starting pool of ca. 10(e15) 80-mer oligonucleotides containing a central region of 40 nucleotides of random sequence. The sequences of ca. 30 cloned aptamers selected after the final SELEX selection round were determined. The protein-binding affinities of seven aptamer candidates were evaluated using a variety of biophysical methods. Result: The bioinformatic analysis of aptamer sequences indicated the presence of various aptamer structures, including G-quadruplex and (TA-rich) non G-quadruplex forms. Results from ELISA-type binding assays revealed that three aptamers specifically bound to the T. denticola protein target with nanomolar affinities. Additional experiments are ongoing. Conclusion: Our preliminary results indicate that two aptamers have significant potential to be used as molecular diagnostic agents for the selective identification of T. denticola strain lineages This abstract is based on research that was funded entirely or partially by an outside source: Research Grants Council of Hong Kong, General Research Fund (#781911)
DescriptionPoster Presentation
Session 76: Clinical Oral Microbiology
Persistent Identifierhttp://hdl.handle.net/10722/201143
ISSN
2015 Impact Factor: 4.602
2015 SCImago Journal Rankings: 1.714

 

DC FieldValueLanguage
dc.contributor.authorGao, Wen_US
dc.contributor.authorYou, Men_US
dc.contributor.authorTanner, JAen_US
dc.contributor.authorLeung, WKen_US
dc.contributor.authorWatt, RMen_US
dc.date.accessioned2014-08-21T07:14:37Z-
dc.date.available2014-08-21T07:14:37Z-
dc.date.issued2014en_US
dc.identifier.citationThe 92nd General Session & Exhibition of the International Association for Dental Research (IADR), Cape Town, South Africa, 25-28 June 2014. In Journal of Dental Research, 2014, v. 93 n. Special issue B: abstract no. 388en_US
dc.identifier.issn0022-0345-
dc.identifier.urihttp://hdl.handle.net/10722/201143-
dc.descriptionPoster Presentation-
dc.descriptionSession 76: Clinical Oral Microbiology-
dc.description.abstractThe oral spirochete bacterium Treponema denticola is implicated in the etiology of periodontal disease. However, the elucidation of more specific disease correlations is hindered by the lack of reliable and discriminatory diagnostic methods for T. denticola cell detection and enumeration. Aptamers are an emerging class of molecular recognition agents, which have binding properties analogous to antibodies. Their potential application for oral microbial cell detection remains to be firmly established. Objective: To identify DNA aptamers against a cell-surface protein from Treponema denticola. Method: A recombinant form of an established cell surface protein from T. denticola ATCC 35405 was used as the target for the identification of single stranded DNA aptamers. A SELEX approach was used for aptamer selection from a starting pool of ca. 10(e15) 80-mer oligonucleotides containing a central region of 40 nucleotides of random sequence. The sequences of ca. 30 cloned aptamers selected after the final SELEX selection round were determined. The protein-binding affinities of seven aptamer candidates were evaluated using a variety of biophysical methods. Result: The bioinformatic analysis of aptamer sequences indicated the presence of various aptamer structures, including G-quadruplex and (TA-rich) non G-quadruplex forms. Results from ELISA-type binding assays revealed that three aptamers specifically bound to the T. denticola protein target with nanomolar affinities. Additional experiments are ongoing. Conclusion: Our preliminary results indicate that two aptamers have significant potential to be used as molecular diagnostic agents for the selective identification of T. denticola strain lineages This abstract is based on research that was funded entirely or partially by an outside source: Research Grants Council of Hong Kong, General Research Fund (#781911)en_US
dc.languageengen_US
dc.publisherSage Publications, Inc. The Journal's web site is located at http://www.sagepub.com/journalsProdDesc.nav?prodId=Journal201925-
dc.relation.ispartofJournal of Dental Researchen_US
dc.rightsJournal of Dental Research. Copyright © Sage Publications, Inc.-
dc.subjectDiagnosis-
dc.subjectMicrobiology-
dc.subjectMolecular biology-
dc.subjectNucleic acids and Periodontal disease-
dc.titleDevelopment of DNA aptamers against Treponema denticolaen_US
dc.typeConference_Paperen_US
dc.identifier.emailTanner, JA: jatanner@hku.hken_US
dc.identifier.emailLeung, WK: ewkleung@hkucc.hku.hken_US
dc.identifier.emailWatt, RM: rmwatt@hku.hken_US
dc.identifier.authorityTanner, JA=rp00495en_US
dc.identifier.authorityLeung, WK=rp00019en_US
dc.identifier.authorityWatt, RM=rp00043en_US
dc.identifier.hkuros233930en_US
dc.identifier.hkuros227282-
dc.identifier.volume93en_US
dc.identifier.issueSpecial issue B: abstract no. 388en_US
dc.publisher.placeUnited States-

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