当前位置: X-MOL 学术J. Aquat. Anim. Health › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Rapid Detection and Monitoring of Flavobacterium psychrophilum in Water by Using a Handheld Field-Portable qPCR System
Journal of Aquatic Animal Health ( IF 1.5 ) Pub Date : 2018-11-22 , DOI: 10.1002/aah.10046
Phong L Nguyen 1 , Ponnerassery S Sudheesh 2 , Austen C Thomas 1 , Mieke Sinnesael 3 , Katherine Haman 4 , Kenneth D Cain 5
Affiliation  

Advances in technology are making it easier for rapid field detection of microbes in aquaculture. Specifically, real-time quantitative PCR (qPCR) analysis, which has traditionally been confined to laboratory-based protocols, is now available in a handheld, field-portable system. The feasibility of using the Biomeme handheld qPCR system for rapid (<50 min) on-site detection and monitoring of Flavobacterium psychrophilum from filtered water samples was evaluated. Paired water samples were collected over a 23-d period from microcosm tanks that housed fish injected with known levels of F. psychrophilum. Water samples were filtered through 0.45-μm nitrocellulose filters and were analyzed with both the Biomeme qPCR platform and a traditional bench qPCR protocol. The two methods identified similar fluctuations in F. psychrophilum DNA throughout the study. Standard curves relating quantification cycles to the number of F. psychrophilum colony-forming units (CFU) were constructed and analyzed; results indicated that CFU increased rapidly between days 6 and 8 of the trial and then progressively decreased during the remaining 15 d. Average calculated log10 (CFU/mL) values were significantly correlated between the two platforms. Rapid, field-based qPCR can be incorporated into daily water quality monitoring protocols to help detect and monitor microbes in aquaculture systems.

中文翻译:

使用手持现场便携式 qPCR 系统快速检测和监测水中的嗜冷黄杆菌

技术的进步使水产养殖中微生物的快速现场检测变得更加容易。具体来说,传统上仅限于基于实验室的协议的实时定量 PCR (qPCR) 分析现在可用于手持、现场便携式系统。评估了使用 Biomeme 手持式 qPCR 系统从过滤水样品中快速(<50 分钟)现场检测和监测嗜冷黄杆菌的可行性。成对的水样是在 23 天的时间内从微型水箱中收集的,该水箱中装有注射了已知水平的嗜冷嗜热菌的鱼。水样通过 0.45-μm 硝酸纤维素过滤器过滤,并使用 Biomeme qPCR 平台和传统的工作台 qPCR 协​​议进行分析。这两种方法确定了 F 的类似波动。整个研究中的嗜冷菌 DNA。构建并分析了将定量周期与嗜冷嗜冷菌菌落形成单位 (CFU) 数量相关的标准曲线;结果表明,CFU 在试验的第 6 天和第 8 天之间迅速增加,然后在剩余的 15 天中逐渐下降。两个平台之间的平均计算 log10 (CFU/mL) 值显着相关。可以将基于现场的快速 qPCR 纳入日常水质监测协议,以帮助检测和监测水产养殖系统中的微生物。两个平台之间的平均计算 log10 (CFU/mL) 值显着相关。可以将基于现场的快速 qPCR 纳入日常水质监测协议,以帮助检测和监测水产养殖系统中的微生物。两个平台之间的平均计算 log10 (CFU/mL) 值显着相关。可以将基于现场的快速 qPCR 纳入日常水质监测协议,以帮助检测和监测水产养殖系统中的微生物。
更新日期:2018-11-22
down
wechat
bug