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| Development and Applification of Real-time Fluorescent Quantitative PCR Assay to Detect Actinobacillus pleuropneumoniae |
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Abstract To develop a Real-time fluoresecent quantitative PCR assay for the detection of Actinobacillus pleuropneumoniae(APP) and apply the developed assay to clinical samples. According to the conserved 391bp neucleotide sequences in apxⅣA gene of APP strains, reported in GenBank, a set of primer pairs were designed to establish APP specie-specific Real-time PCR and standard curve occurred after amplification using gradient diluted plasmid as template. After developed, the method was used to quantitatively detect the copies of APP in lung tissues both collected from disease or artificially infected pigs. Data obtained demonstrated that the newly developed method was highly sensitive, specie-specific, repeatable and applicable for clinical diagnosis. The method established in this research has potential to pave the way for early and rapid detection of APP or quantitative analysis for the infection degree of APP.
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Received: 29 June 2009
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