视觉LAMP法检测对虾和小龙虾白斑综合征病毒

IF 1.5 4区 农林科学 Q3 FISHERIES Turkish Journal of Fisheries and Aquatic Sciences Pub Date : 2022-06-17 DOI:10.4194/trjfas21519
Chao Ma, Z. Tian, Lu Bai, Lili Yang, Jijuan Cao
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引用次数: 0

摘要

白斑综合征病毒(WSSV)给水产养殖业造成了巨大的经济损失,迄今为止还没有针对这种致命病毒的有效治疗方法。因此,对虾白斑综合征病毒(WSSV)的快速检测至关重要。本研究以环介导等温扩增(LAMP)技术为基础,建立对虾WSSV的快速视觉检测方法。采用荧光扩增曲线(SYTO-9荧光染料)和LAMP可视化(钙黄素)观察扩增产物的颜色变化,分析方法的特异性、灵敏度和重复性。结果表明,该方法可特异性检测WSSV,检测灵敏度为1 fg/μL,具有良好的重现性和可靠性。该方法用于养殖虾和小龙虾样品的分析;329份疑似临床标本中,WSSV阳性诊断率为27.96%。温度为20 ~ 26℃时,WSSV侵染活性最高。基于lamp的WSSV快速视觉检测方法不需要复杂的实验仪器,且有稳定热源的设备就足以进行反应。它易于使用,可用于对虾养殖场。
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Visual LAMP Method for Detection of White Spot Syndrome Virus in Shrimp and Crayfish
White spot syndrome virus (WSSV) has caused large economic losses to the aquaculture industry, and thus far there are no efficient therapeutic treatments available against this lethal virus. Therefore, rapid detection of white spot syndrome virus (WSSV) in shrimp is crucial. This study was conducted to establish a rapid visual detection method for WSSV in shrimp and on the basis of loop-mediated isothermal amplification (LAMP). The color change of the amplification products was observed using a fluorescence amplification curve (with the SYTO-9 fluorescent dye) and LAMP visualization (with calcein), and the method`s specificity, sensitivity, and reproducibility were analyzed. It showed that this method can specifically detect WSSV, with a detection sensitivity of 1 fg/μL and has good reproducibility and reliability. This method was used to analyze cultured shrimp and crayfish samples; among 329 suspected clinical samples, the positive diagnostic rate of WSSV was 27.96%. The infection activity of WSSV was the highest at temperatures of 20–26°C. The LAMP-based method for rapid visual detection of WSSV does not need complex experimental instruments, and an equipment with a stable heat source is sufficient for the reaction. It is easy to use and can be used at shrimp breeding sites.
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来源期刊
Turkish Journal of Fisheries and Aquatic Sciences
Turkish Journal of Fisheries and Aquatic Sciences FISHERIES-MARINE & FRESHWATER BIOLOGY
CiteScore
3.10
自引率
0.00%
发文量
43
审稿时长
3 months
期刊介绍: Turkish Journal of Fisheries and Aquatic Sciences" (TrJFAS) is a refereed academic journal has been published by Central Fisheries Research Institute of Turkey and Japan International Cooperation Agency (JICA), and published in English. It aims to address research and needs of all working and studying within the many varied areas of fisheries and aquatic sciences. The Journal publishes English language original research papers, critical review articles, short communications and technical notes on applied or scientific research relevant to freshwater, brackish and marine environments. TrJFAS was published biannually (April & November) between 2001 and 2009. A great number of manuscripts have been submitted to the journal for review from acceptance of the SCI index. Thereby, the journal has been published quarterly (March, June, September and December) from 2010 to 2017. The journal will be published monthly in 2018.
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