Establishment of SYBR Green I Real-Time PCR for Detection of Streptococcus agalactiae in Aquaculture Waters

IF 0.7 4区 农林科学 Q4 FISHERIES Journal of Applied Ichthyology Pub Date : 2023-10-20 DOI:10.1155/2023/9199300
Zaiyong Si, Zhenhua Ma, Fuguang Luo, Jingu Shi, Jie Huang, Zhiqiang Wang, Yanhong Wen, Yi Yi, Shuyu Han
{"title":"Establishment of SYBR Green I Real-Time PCR for Detection of Streptococcus agalactiae in Aquaculture Waters","authors":"Zaiyong Si, Zhenhua Ma, Fuguang Luo, Jingu Shi, Jie Huang, Zhiqiang Wang, Yanhong Wen, Yi Yi, Shuyu Han","doi":"10.1155/2023/9199300","DOIUrl":null,"url":null,"abstract":"Streptococcus agalactiae has a serious negative impact on tilapia aquaculture, and rapid detection of trace S. agalactiae in aquaculture waters and timely and effective sterilization measures could significantly reduce the probability of its outbreak in tilapia farming. Here, we established a fluorescence quantitative detection method of S. agalactiae in tilapia aquaculture waters based on the Streptococcus agalactiae CAMP factor (cfb) (GU217532.1) sequence of its CAMP factor. The results showed that the Ct value and logarithm of plasmid copy number presented a good linear relationship (y = −3.49x + 38.78; R2 = 0.997) in the plasmid concentration range of 1.57 × 102−1.57 × 109 copies/μL, and the lowest concentration for sensitive detection of the established method was 1.57 × 102 copies/μL. The method exhibited high specificity for the detection of S. agalactiae and generated negative results when using the DNA of Aeromonas veronii, Staphylococcus epidermidis, Aeromonas hydrophila, and Edwardsiella tarda as templates for qPCR. Intra- and inter-group repeated experiments produced variation coefficients lower than 2%, indicating high stability and specificity of the method. The method was then used to detect S. agalactiae in two tilapia aquaculture waters. The results showed that the concentration of S. agalactiae was 12 copies/mL and 2 copies/mL in the two water samples, respectively. The method can directly detect trace S. agalactiae in the aquaculture water and facilitate the diagnosis, prevention, and control of S. agalactiae in tilapia aquaculture, which will greatly reduce the probability of large-scale outbreak of S. agalactiae and economic loss.","PeriodicalId":14894,"journal":{"name":"Journal of Applied Ichthyology","volume":"14 12","pages":"0"},"PeriodicalIF":0.7000,"publicationDate":"2023-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Applied Ichthyology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1155/2023/9199300","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"FISHERIES","Score":null,"Total":0}
引用次数: 0

Abstract

Streptococcus agalactiae has a serious negative impact on tilapia aquaculture, and rapid detection of trace S. agalactiae in aquaculture waters and timely and effective sterilization measures could significantly reduce the probability of its outbreak in tilapia farming. Here, we established a fluorescence quantitative detection method of S. agalactiae in tilapia aquaculture waters based on the Streptococcus agalactiae CAMP factor (cfb) (GU217532.1) sequence of its CAMP factor. The results showed that the Ct value and logarithm of plasmid copy number presented a good linear relationship (y = −3.49x + 38.78; R2 = 0.997) in the plasmid concentration range of 1.57 × 102−1.57 × 109 copies/μL, and the lowest concentration for sensitive detection of the established method was 1.57 × 102 copies/μL. The method exhibited high specificity for the detection of S. agalactiae and generated negative results when using the DNA of Aeromonas veronii, Staphylococcus epidermidis, Aeromonas hydrophila, and Edwardsiella tarda as templates for qPCR. Intra- and inter-group repeated experiments produced variation coefficients lower than 2%, indicating high stability and specificity of the method. The method was then used to detect S. agalactiae in two tilapia aquaculture waters. The results showed that the concentration of S. agalactiae was 12 copies/mL and 2 copies/mL in the two water samples, respectively. The method can directly detect trace S. agalactiae in the aquaculture water and facilitate the diagnosis, prevention, and control of S. agalactiae in tilapia aquaculture, which will greatly reduce the probability of large-scale outbreak of S. agalactiae and economic loss.
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
水产养殖水体无乳链球菌实时荧光定量PCR检测方法的建立
无乳链球菌对罗非鱼养殖有严重的负面影响,快速检测养殖水体中微量无乳链球菌并采取及时有效的灭菌措施可显著降低其在罗非鱼养殖中爆发的概率。本研究基于无乳链球菌CAMP因子(cfb) (GU217532.1)序列,建立了罗非鱼养殖水体中无乳链球菌的荧光定量检测方法。结果表明,Ct值与质粒拷贝数的对数呈良好的线性关系(y =−3.49x + 38.78;R2 = 0.997),质粒浓度范围为1.57 × 102 ~ 1.57 × 109 copies/μL,建立的方法灵敏检测的最低浓度为1.57 × 102 copies/μL。该方法检测无乳链球菌的特异性高,以维罗氏气单胞菌、表皮葡萄球菌、嗜水气单胞菌和迟发爱德华菌的DNA为模板进行qPCR时均为阴性。组内和组间重复实验变异系数均小于2%,表明该方法具有较高的稳定性和特异性。应用该方法对两种罗非鱼养殖水体中的无乳链球菌进行了检测。结果表明,两种水样中无乳链球菌的浓度分别为12拷贝/mL和2拷贝/mL。该方法可直接检测出养殖水体中痕量无乳链球菌,便于罗非鱼养殖中无乳链球菌的诊断、防治,将大大降低无乳链球菌大规模爆发的概率和经济损失。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
Journal of Applied Ichthyology
Journal of Applied Ichthyology 生物-海洋与淡水生物学
CiteScore
2.30
自引率
11.10%
发文量
73
审稿时长
3-8 weeks
期刊介绍: The Journal of Applied Ichthyology publishes articles of international repute on ichthyology, aquaculture, and marine fisheries; ichthyopathology and ichthyoimmunology; environmental toxicology using fishes as test organisms; basic research on fishery management; and aspects of integrated coastal zone management in relation to fisheries and aquaculture. Emphasis is placed on the application of scientific research findings, while special consideration is given to ichthyological problems occurring in developing countries. Article formats include original articles, review articles, short communications and technical reports.
期刊最新文献
Population Size Structure and Length–Weight Relationships of Selected Pelagic Fishes From The Gambian Waters (West Africa) DNA Barcoding of Nematode Parasites Infecting Ompok bimaculatus and Nemacheilus anguilla From Barvi Reservoir, Maharashtra Unraveling the Impact of Climate Change on Fish Physiology: A Focus on Temperature and Salinity Dynamics Length-Weight Relationships of Native and Non-Native Fishes in the Lower Red River Catchment, USA Assessing Invasive Carp in the Neosho River-Grand Lake System of Kansas and Oklahoma
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1