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Development of a DNA probe and PCR assay for the detection of Actinobacillus pleuropneumoniae

Posted on:1993-03-01Degree:M.SType:Thesis
University:University of Missouri - ColumbiaCandidate:Thornton, Kathryn AnnFull Text:PDF
GTID:2473390014495306Subject:Biology
Abstract/Summary:
A DNA probe was created for the detection of Actinobacillus pleuropneumoniae DNA from a genomic library of BamHI digested A. pleuropneumoniae serotype 2 (27089) DNA cloned in the BamHI site of the plasmid vector pTZ18U. The probe, pTA4, contains a 3.1 Kb insert of A. pleuropneumoniae and is specific in its affinity for all serotypes of A. pleuropneumoniae analyzed. DNA sequences were obtained from the 3.1 Kb insert. Five primers were designed for use in the polymerase chain reaction. DNA amplification of purified DNA from all serotypes A. pleuropneumoniae analyzed was achieved. Preliminary evidence indicates that amplification of Haemophilus parasuis, Bordetella bronchiseptica, Escherichia coli, and leukocytic swine DNA is unlikely and that the amplification of P. multocida DNA could be eliminated with two specific combination of the five primers.
Keywords/Search Tags:DNA probe, Pleuropneumoniae, Five primers, Biology
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