基于核酸适配体的PCR法检测溶藻弧菌及其灭活菌
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1. 鳗鲡现代产业技术教育部工程研究中心, 福建省海洋渔业资源与生态环境重点实验室, 集美大学水产学院, 福建 厦门 361021;
2. 广东雅琪生物科技股份有限公司, 广东 广州 510000;
3. 厦门阳嘉环保科技有限公司, 福建 厦门 361021

作者简介:

汤学敏(1989-),女,硕士,从事微生物及微生态制剂方面的研究.E-mail:esta_tang@163.com

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中图分类号:

S947;Q939.9

基金项目:

国家海洋经济发展区域示范项目(14PYY050SF03);福建省自然科学基金项目(2018J01455);鳗鲡现代产业技术教育部工程研究中心开放基金项目(RE201808);大学生创新创业训练计划项目(201710390021,201810390073,2018lxj025,2018lxj058,2018lxj066,2018lxj067).


Detection of Vibrio alginolyticus and its inactivated form based on aptamers and polymerase chain reaction
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1. Engineering Research Center of the Modern Technology for Eel Industry, Ministry of Education;Fujian Provincial Key Laboratory of Marine Fishery, Resources and Eco-environment, Fisheries College of Jimei University, Xiamen 361021, China;
2. Guangdong Atech Biotechnology Limited Corporation, Guangzhou 510000, China;
3. Xiamen Yangjia Environmental Science and Technology Limited Company, Xiamen 361021, China

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    摘要:

    溶藻弧菌()分布广,数量多,发病率高,是水产养殖中常见的条件致病菌,而对溶藻弧菌进行快速准确的识别鉴定是其病害防治的前提和基础。核酸适配体,因为具有较高的亲和特异性,在微生物的识别鉴定方面展现出了巨大的优势。本文利用核酸适配体和适配体筛选产物,通过结合、洗涤、加热分离、PCR扩增以及电泳检测等步骤,对溶藻弧菌进行了检测鉴定。结果表明,适配体和筛选产物都能对溶藻弧菌及其灭活菌进行较好的识别鉴定,适配体筛选产物对溶藻弧菌的检测下限为103 cfu/mL,而对其灭活菌的检测下限为102 cfu/mL,适配体对溶藻弧菌及其灭活菌的检测下限都可达到10 cfu/mL。该方法对溶藻弧菌有较好的亲和特异性,并能较好地区分溶藻弧菌与哈维氏弧菌等水产常见病原菌,在水产病害的检测中显示了较好的应用前景。

    Abstract:

    is a common conditional pathogen in aquaculture, which is widely distributed in the world. Rapid detection of the pathogen is necessary for the prevention and control of the diseases caused by the bacterium. Aptamers exhibit great advantages in the detection of microorganisms owing to their high affinities and specificities. In the present study, was detected based on its aptamer and its selection products following various steps, such as binding, washing, separation, amplification, and electrophoresis. The results showed that the aptamer and the selection products could superiorly detect and its inactivated form. The lowest detection limits were 10 cfu/mL of cfu/mL of inactivated by the selection products. The system of detection had better affinity and specificity for from other aquatic pathogens such as , and therefore shows good application potential in the detection of pathogens in aquaculture.

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汤学敏,林文燕,鄢庆枇,刘慧敏,彭英林,赵铨,林旭俊,郑江.基于核酸适配体的PCR法检测溶藻弧菌及其灭活菌[J].中国水产科学,2019,26(2):391-396
TANG Xuemin, LIN Wenyan, YAN Qinpi, LIU Huimin, PENG Yinglin, ZHAO Quan, LIN Xujun, ZHENG Jiang. Detection of Vibrio alginolyticus and its inactivated form based on aptamers and polymerase chain reaction[J]. Journal of Fishery Sciences of China,2019,26(2):391-396

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  • 在线发布日期: 2019-03-27
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