Guangxi Key Laboratory of Aquatic Genetic Breeding and Healthy Aquaculture, Academy of Fishery Sciences, Nanning, Guangxi 530021, China.
Qingdao Customs, Qingdao 266000, China.
J Virol Methods. 2019 Nov;273:113683. doi: 10.1016/j.jviromet.2019.113683. Epub 2019 Jul 5.
This study aimed to establish pyrosequencing methods to detect white spot syndrome virus (WSSV). One pair of polymerase chain reaction (PCR) primers, and one pyrosequencing primer, were designed for WSSV. The pyrosequencing reaction system and conditions were optimized and a pyrosequencing method for detecting WSSV was successfully established. This method was able to specifically detect WSSV in eight viruses, with high sensitivity. The minimum detectable limit for nucleic acid was 23 copies/μL. The method was verified by detecting WSSV in 1881 batches of samples collected from domestic and imported shrimps. The detection results were more sensitive than conventional PCR. This research has therefore provided a new detection method for monitoring, and controlling aquatic animal virus diseases.
本研究旨在建立焦磷酸测序方法来检测白斑综合征病毒 (WSSV)。设计了一对用于 WSSV 的聚合酶链反应 (PCR) 引物和一个焦磷酸测序引物。优化了焦磷酸测序反应体系和条件，成功建立了检测 WSSV 的焦磷酸测序方法。该方法能够特异性检测 8 种病毒中的 WSSV，具有较高的灵敏度。核酸的最小检测限为 23 拷贝/μL。该方法通过检测从国产和进口虾中采集的 1881 批样品中的 WSSV 进行了验证。检测结果比常规 PCR 更敏感。因此，本研究为监测和控制水生动物病毒病提供了一种新的检测方法。
