Journal
ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
Volume 29, Issue 17, Pages 24696-24703Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s11356-021-17536-w
Keywords
Karenia mikimotoi; Loop-mediated isothermal amplification (LAMP); Lateral flow dipstick (LFD); Rapid detection
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Funding
- Special Foundation for National Science and Technology Basic Research Program of China [2018FY100207]
- National Key Research and Development Program of China [2017YFC1404304]
- Strategic Priority Research Program of the Chinese Academy of Sciences [XDA23050302]
- Sino-Australian Centre for Healthy Coasts [2016YFE0101500]
- Aoshan Science and Technology Innovation Project, Qingdao National Laboratory for Marine Science and Technology [2016ASKJ02-1]
- Foundation of Key Laboratory of Integrated Marine Monitoring
- Applied Technologies for Harmful Algal Blooms [MATHAB201707]
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A LAMP method coupled with a LFD was developed for detecting harmful algae Karenia mikimotoi, showing high sensitivity and specificity, indicating a promising approach for detecting and monitoring harmful algal blooms.
Harmful algal blooms frequently occur in various coastal regions worldwide, deteriorating marine ecology and causing huge economic losses. Therefore, developing a potential method for rapid detection of harmful algae species is highly necessitated. In this study, a loop-mediated isothermal amplification (LAMP) method coupled with a lateral flow dipstick (LFD) was developed for detecting the harmful algae Karenia mikimotoi. In this method, the internal transcribed spacer (ITS) sequence of K. mikimotoi was used as the template, and the corresponding specific primers were designed by the online software PrimerExplorer V5. Biotin was labeled on the 5' end of forward inner primer (FIP), and the LAMP reaction was performed under the determined optimal conditions of 63 degrees C and 60 min. The lowest concentration of K. mikimotoi DNA tested using LAMP was 3.3 x 10(-1) pg/mu L. Additionally, a 6-FAM-labeled probe was designed and displayed on the LFD after hybridization of the amplified product with the probe. The results demonstrated that LAMP-LFD could be a promising approach for detecting and monitoring harmful algae due to its high sensitivity and specificity.
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