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针对近年来由虹彩病毒所引起的海水养殖鱼类疾病呈日趋严重的态势,该文在证实了引发我国大黄鱼大规模流行病的病原为一种虹彩病毒及测定病毒全基因组序列(111,760bp;GenBankaccessionnumber:AY779031)的基础上,通过与已报道虹彩病毒核酸序列进行分析比较,结合生物信息学手段,确定了以虹彩病毒ATPase基因保守区序列(295bp)作为扩增靶序列,设计合成了一对特异性引物,通过改进PCR模板的制备方法和优化扩增条件,建立了大黄鱼虹彩病毒PCR快速检测技术,并开发成简便、快速、实用的检测试剂盒,该试剂盒的检测灵敏度相当于30个病毒粒子,模板制备时间约30min、回收率为52%、半个工作日即可得到准确的结果,无非特异性扩增带,适用于大黄鱼虹彩病毒病的早期快速诊断、苗种的检疫及水质环境的监测,目前正在推广应用。
In view of the increasingly serious situation of mariculture fish diseases caused by iridescent virus in recent years, this article confirms that the causative agent of large-scale epidemic disease of Chinese big yellow croaker is an iridescent virus and the whole genome sequence of the virus (111,760 bp; GenBank accession number: AY779031). Based on the analysis and comparison with reported nucleotide sequences of iris virus, bioinformatics methods were used to determine the sequence of the conserved region of ATPase gene (295 bp) Specific primers, PCR rapid detection method was established by improving the preparation method of PCR template and optimizing amplification conditions, and developed into a simple, rapid and practical detection kit, the detection sensitivity of the kit is equivalent to 30 A virus particle, template preparation time of about 30min, the recovery rate was 52%, half a working day to get accurate results, no non-specific amplification band, suitable for rapid detection of large yellow croaker iris virus disease, quarantine of seedlings And water quality and environmental monitoring, is now promoting the application.