Liu Wenyue, Zhang Guangying, Xu Di, Ye Jingqin, Lu Ying
College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China.
Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai 201306, China.
Foods. 2023 Sep 28;12(19):3605. doi: 10.3390/foods12193605.
can cause disease in aquatic animals and humans, therefore, rapid and simple field detection of pathogenic is important for early disease prevention. In this study, a novel recombinase-aided amplification (RAA) combined test strip with double T-lines (RAA-TS-DTL) was developed for the rapid detection of in aquatic products. Pathogenic was detected using the virulence gene and the housekeeping gene gene as the dual target of the test strip. The RAA-TS-DTL method showed 100% specificity for , and no cross-reaction was observed with spp. or other bacteria ( = 14). Furthermore, sensitive detection of in oysters was achieved. The LODs of the and genes were 6 CFU/mL and 23 CFU/mL, respectively, which was about five times higher than that of the commercial test strip. The method was validated with spiked samples ( = 60) of fish, shrimp and oyster. The consistency between RAA-TS-DTL and the traditional culture method was 97.9%. In addition, the entire process of detection, including preparation of the sample, could be completed within 50 min. Our results indicated that the developed RAA-TS-DTL was a reliable and useful tool for rapid screening or on-site detection of pathogenic in aquatic products and aquaculture water.
可导致水生动物和人类患病，因此，对病原体进行快速、简单的现场检测对于早期疾病预防至关重要。在本研究中，开发了一种新型的重组酶辅助扩增（RAA）结合双T线试纸条（RAA-TS-DTL）用于快速检测水产品中的[病原体名称未给出]。使用毒力[基因名称未给出]基因和管家基因[基因名称未给出]基因作为试纸条的双重靶标来检测致病性[病原体名称未给出]。RAA-TS-DTL方法对[病原体名称未给出]显示出100%的特异性，与[其他菌属名称未给出] spp. 或其他细菌（n = 14）未观察到交叉反应。此外，实现了对牡蛎中[病原体名称未给出]的灵敏检测。[基因名称未给出]基因和[基因名称未给出]基因的检测限分别为6 CFU/mL和23 CFU/mL，这比商业试纸条的检测限高约五倍。该方法用鱼、虾和牡蛎的加标样品（n = 60）进行了验证。RAA-TS-DTL与传统培养方法之间的一致性为97.9%。此外，包括样品制备在内的整个检测过程可在50分钟内完成。我们的结果表明，所开发的RAA-TS-DTL是一种可靠且有用的工具，可用于快速筛查或现场检测水产品和养殖水中的致病性[病原体名称未给出]。
