| Summary: | 碩士 === 國立中興大學 === 生物科技學研究所 === 97 === Infectius bursa disease (IBD) is a highly contagious disease in young chicken. The pathogen of IBD is infectious bursal disease virus (IBDV). Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method with advantages of simplicity, rapidity, and specificity. The amplification depends on a set of special primer combined with a DNA polymerase with strand replacement activity, and large amounts of products were synthesized at isothermal condition in one hour. In this study, LAMP combined with reverse transcription for detection of IBDV was performed. The initial assay confirmed the primer set amplified correct target. Extracted genomic RNA of IBDV was used for the sensitivity test of RT-LAMP. The limit of detection was determined to be 0.01 fg RNA of IBDV in the present study. In the specificity test, LAMP only amplified sample contained IBDV; CAV and AIV were negative results. In the RT-LAMP assay with clinical samples, the IBDV in bursa samples detected correctly. Moreover, the LAMP method and reverse transcription worked at the same time that more convenient than conventional RT-PCR. These data suggest the RT-LAMP method is a potential diagnosis tool for detecting IBDV.
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