Duan Xinbing, Li Jiji, Shi Hui, Tao Zhen, Wei Xuelian, Ye Yingying, Guo Baoying
Marine Science and Technology College, Zhejiang Ocean University, Zhoushan 316022, China.
National Engineering Research Center for Marine Aquaculture, Zhejiang Ocean University, Zhoushan 316022, China.
Animals (Basel). 2024 May 10;14(10):1427. doi: 10.3390/ani14101427.
The visceral white nodules disease in the internal organs of has caused significant harm in the aquaculture of this species, with considered one of the core pathogens causing this disease. In this study, we designed three pairs of specific nested PCR primers targeting the gene of , a crucial component of the Type III secretion system (T3SS), which is instrumental in bacterial pathogenesis and virulence. Through the optimization of PCR reaction conditions, specificity testing, and sensitivity determination, a method was established for the accurate detection of . This method yielded single amplification products, exhibited a false positive rate of zero for reference bacteria, and achieved a detection sensitivity of a minimum of 2.62 copies/reaction for the target sequence. Using the detection method, we conducted analyses on the diseased populations of , involving a total of 64 screened fishes along the southeast coast of China from 2021 to 2023. The results revealed that the infection rate of in diseased exceeded over 90% in March and April, while in other months, the maximum recorded infection rate was merely 10%. The detection method developed in this study shows potential for early warning and routine monitoring of visceral white nodules disease in the internal organs of species such as .
该物种内脏白色结节病已对其水产养殖造成严重危害，[病原体名称]被认为是引发该病的核心病原体之一。在本研究中，我们针对[病原体名称]的Ⅲ型分泌系统（T3SS）的关键组成部分[基因名称]设计了三对特异性巢式PCR引物，该系统在细菌致病机制和毒力方面发挥着重要作用。通过优化PCR反应条件、进行特异性测试和灵敏度测定，建立了一种准确检测[病原体名称]的方法。该方法产生单一扩增产物，对参考细菌的假阳性率为零，对目标序列的检测灵敏度达到最低2.62拷贝/反应。利用该检测方法，我们对[物种名称]的患病群体进行了分析，涉及2021年至2023年在中国东南沿海筛选的64条鱼。结果显示，患病[物种名称]中[病原体名称]的感染率在3月和4月超过90%，而在其他月份，记录到的最高感染率仅为10%。本研究开发的检测方法在[物种名称]等物种内脏白色结节病的早期预警和常规监测方面显示出潜力。
