CAJ | 학술논문

本研究利用Primer Explorer V4软件设计了针对鸡传染性法氏囊病毒（IBDV）VP1基因保守区6个特异性部位的4条引物，建立了IBDV的反转录环媒恒温检测方法。该方法反应体系在恒温水浴锅中作用1 h即可得到其特有的阶梯状条带，加入荧光素后肉眼可直接观察结果。本方法特异性高，对IBDV的最低检出量为10-6稀释倍数，敏感性是普通RT-PCR的100倍。反转录环媒恒温检测方法和普通RT-PCR方法检测临床样品的符合率为93.8%。因此该方法可以作为IBDV的综合防治和疾病诊断的实用方法。
본연구이용Primer Explorer V4연건설계료침대계전염성법씨낭병독（IBDV）VP1기인보수구6개특이성부위적4조인물，건립료IBDV적반전록배매항온검측방법。해방법반응체계재항온수욕과중작용1 h즉가득도기특유적계제상조대，가입형광소후육안가직접관찰결과。본방법특이성고，대IBDV적최저검출량위10-6희석배수，민감성시보통RT-PCR적100배。반전록배매항온검측방법화보통RT-PCR방법검측림상양품적부합솔위93.8%。인차해방법가이작위IBDV적종합방치화질병진단적실용방법。
The objective of this study is to develop a rapid and sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the detection of infectious bursal disease virus (IBDV). Six primers were designed to amplify the VP1 gene of IBDV using PrimerExplor-er V4. The optimal reaction condition of the current RT-LAMP for IBDV was 65 ℃ for 60 mins. It was capable of detecting IBDV from clinical samples and differentiating IBDV from other avain vi-ruses, and does not require an additional expensive equipment. The minimum detection limit of the RT-LAMP assay was 10-6, making this assay approximately 100-fold higher than that of one-step RT-PCR. The assay were evaluated by comparison with RT-PCR, by 93.8% consistence. The specificity and simplicity of the assay could make it a useful method for the detection of IBDV infection.