Lin Hung-Yun, Yen Shao-Chieh, Tsai Shou-Kuan, Shen Fan, Lin John Han-You, Lin Han-Jia
Center of Excellence for the Oceans, National Taiwan Ocean University, Keelung 20224, Taiwan.
Department of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung 20224, Taiwan.
Life (Basel). 2022 Feb 13;12(2):276. doi: 10.3390/life12020276.
Infectious diseases are considered the greatest threat to the modern high-density shrimp aquaculture industry. Specificity, rapidity, and sensitivity of molecular diagnostic methods for the detection of asymptomatic infected shrimp allows preventive measures to be taken before disease outbreaks. Routine molecular detection of pathogens in infected shrimp can be made easier with the use of a direct polymerase chain reaction (PCR). In this study, four direct PCR reagent brands were tested, and results showed that the detection signal of direct PCR in hepatopancreatic tissue was more severely affected. In addition, portable capillary electrophoresis was applied to improve sensitivity and specificity, resulting in a pathogen detection limit of 25 copies/PCR-reaction. Juvenile shrimp from five different aquaculture ponds were tested for white spot syndrome virus infection, and the results were consistent with the Organization for Animal Health's certified standard method. Furthermore, this methodology could be used to examine single post larvae shrimp. The overall detection time was reduced by more than 58.2%. Therefore, the combination of direct PCR and capillary electrophoresis for on-site examination is valuable and has potential as a suitable tool for diagnostic, epidemiological, and pathological studies of shrimp aquaculture.
传染病被认为是现代高密度对虾养殖业面临的最大威胁。用于检测无症状感染对虾的分子诊断方法具有特异性、快速性和敏感性，这使得在疾病爆发前能够采取预防措施。使用直接聚合酶链反应（PCR）可以使感染对虾中病原体的常规分子检测更加容易。在本研究中，对四个直接PCR试剂品牌进行了测试，结果表明肝胰腺组织中直接PCR的检测信号受到的影响更为严重。此外，应用便携式毛细管电泳来提高灵敏度和特异性，病原体检测限达到25拷贝/PCR反应。对来自五个不同养殖池塘的幼虾进行了白斑综合征病毒感染检测，结果与世界动物卫生组织认证的标准方法一致。此外，该方法可用于检测单个仔虾。总体检测时间减少了58.2%以上。因此，直接PCR和毛细管电泳相结合用于现场检测具有重要价值，有潜力成为对虾养殖诊断、流行病学和病理学研究的合适工具。
