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我国水禽中流行的禽1型副黏病毒(APMV-1)除基因Ⅶ外,基因Ⅸ型APMV-1日渐增多,且呈现基因型多样性。本研究基于鸭源基因Ⅸ型APMV-1的F基因特异性序列,建立了一种可快速检测基因Ⅸ型APMV-1的实时荧光定量RT-PCR方法。该方法仅能特异性检出基因Ⅸ型APMV-1,其检测敏感性高达100拷贝/μL,重复试验批间变异系数小于5%;应用该方法对人工感染鸭组织和采集的临床样品分别进行检测,其结果与常规RT-PCR检测、病毒分离结果的符合率分别为100.0%,96.1%和90.1%,81.8%。以上结果表明,本研究建立的基因Ⅸ型APMV-1实时荧光定量RT-PCR检测方法特异性强、敏感性高、重复性好,为该病临床样品检测提供了快速有效的手段,适于鸭源基因Ⅸ型APMV-1感染的病原学监测。
In addition to the gene Ⅶ, the avian Paramyxovirus-1 Paramyxovirus Type 1 (APMV-1), which is prevalent in waterfowl of our country, is experiencing an increased genotype of APMV-1. In this study, a real-time fluorescence quantitative RT-PCR method for rapid detection of gene Ⅸ APMV-1 was established based on the F gene-specific sequence of duck origin gene Ⅸ APMV-1. The method can only detect gene type APMV-1 specifically, its detection sensitivity is as high as 100 copies / μL, and the coefficient of variation between repeated experiments is less than 5%. The method is applied to separately inoculate duck tissues and clinical samples collected The results were in agreement with the results of routine RT-PCR and virus isolation, which were 100.0%, 96.1% and 90.1%, 81.8% respectively. The above results show that the method of real-time fluorescence quantitative RT-PCR of APMV-1 gene Ⅸ established in this study has high specificity, high sensitivity and good repeatability, which provides a quick and effective method for clinical sample detection of the disease and is suitable for duck Etiological surveillance of APMV-1 genotype Ⅸ infection.