Species-specific DNA probes for the identification of Actinobacillus actinomycetemcomitans

 

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dc.contributor.advisor Steyn, Lafras M en_ZA
dc.contributor.author Emanuel, Margot en_ZA
dc.date.accessioned 2017-08-23T13:12:39Z
dc.date.available 2017-08-23T13:12:39Z
dc.date.issued 1990 en_ZA
dc.identifier.citation Emanuel, M. 1990. Species-specific DNA probes for the identification of Actinobacillus actinomycetemcomitans. University of Cape Town. en_ZA
dc.identifier.uri http://hdl.handle.net/11427/24965
dc.description.abstract A DNA probe was developed for the identification of the periodontal pathogen, Actinobacillus actinomycetemcomitans. Chromosomal DNA was extracted from A. actinomycetemcomitans, digested with a restriction enzyme, Sau3A, ligated to plasmid DNA (pUC18) and transformed into JM109 cells to give a partial A. actinomycetemcomitans library. The library was screened using Southern blot analysis. Out of the nine inserts tested, one was found to be species specific as it did not cross-hybridise to Haemophilus aphrophilus, a closely related organism which occurs in the normal oral microflora, nor did it cross-hybridise with 7 species of Bacteroides tested. A level of detection of 104 cells or 50ng of A. actinomycetemcomitans was obtained. The probe has a length of 779bp and out of 30 restriction enzymes tested, only SspI was found to have a restriction site in the insert. The probe was tested on clinical specimens obtained from five different periodontitis patient groups and was shown to correlate with culture results in eighteen out of twenty-two cases in detecting A. actinomycetemcomitans. en_ZA
dc.language.iso eng en_ZA
dc.subject.other Actinobacillus - pathogenicity~DNA Probes - analysis~Periodontitis - etiology en_ZA
dc.title Species-specific DNA probes for the identification of Actinobacillus actinomycetemcomitans en_ZA
dc.type Master Thesis
dc.date.updated 2017-07-10T12:42:58Z
uct.type.publication Research en_ZA
uct.type.resource Thesis en_ZA
dc.publisher.institution University of Cape Town
dc.publisher.faculty Faculty of Health Sciences en_ZA
dc.publisher.department Division of Medical Microbiology en_ZA
dc.type.qualificationlevel Masters
dc.type.qualificationname MSc (Med) en_ZA
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uct.type.filetype Text
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dc.identifier.apacitation Emanuel, M. (1990). <i>Species-specific DNA probes for the identification of Actinobacillus actinomycetemcomitans</i>. (Thesis). University of Cape Town ,Faculty of Health Sciences ,Division of Medical Microbiology. Retrieved from http://hdl.handle.net/11427/24965 en_ZA
dc.identifier.chicagocitation Emanuel, Margot. <i>"Species-specific DNA probes for the identification of Actinobacillus actinomycetemcomitans."</i> Thesis., University of Cape Town ,Faculty of Health Sciences ,Division of Medical Microbiology, 1990. http://hdl.handle.net/11427/24965 en_ZA
dc.identifier.vancouvercitation Emanuel M. Species-specific DNA probes for the identification of Actinobacillus actinomycetemcomitans. [Thesis]. University of Cape Town ,Faculty of Health Sciences ,Division of Medical Microbiology, 1990 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/24965 en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Emanuel, Margot AB - A DNA probe was developed for the identification of the periodontal pathogen, Actinobacillus actinomycetemcomitans. Chromosomal DNA was extracted from A. actinomycetemcomitans, digested with a restriction enzyme, Sau3A, ligated to plasmid DNA (pUC18) and transformed into JM109 cells to give a partial A. actinomycetemcomitans library. The library was screened using Southern blot analysis. Out of the nine inserts tested, one was found to be species specific as it did not cross-hybridise to Haemophilus aphrophilus, a closely related organism which occurs in the normal oral microflora, nor did it cross-hybridise with 7 species of Bacteroides tested. A level of detection of 104 cells or 50ng of A. actinomycetemcomitans was obtained. The probe has a length of 779bp and out of 30 restriction enzymes tested, only SspI was found to have a restriction site in the insert. The probe was tested on clinical specimens obtained from five different periodontitis patient groups and was shown to correlate with culture results in eighteen out of twenty-two cases in detecting A. actinomycetemcomitans. DA - 1990 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 1990 T1 - Species-specific DNA probes for the identification of Actinobacillus actinomycetemcomitans TI - Species-specific DNA probes for the identification of Actinobacillus actinomycetemcomitans UR - http://hdl.handle.net/11427/24965 ER - en_ZA


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