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DOI | 10.1016/j.hal.2019.01.011 |
Rapid and sensitive detection method for Karlodinium veneficum by recombinase polymerase amplification coupled with lateral flow dipstick | |
Fu, Mengqi1; Chen, Guofu1; Zhang, Chunyun1,2; Wang, Yuanyuan1; Sun, Rui1; Zhou, Jin3 | |
发表日期 | 2019 |
ISSN | 1568-9883 |
EISSN | 1878-1470 |
卷号 | 84页码:1-9 |
英文摘要 | The dinoflagellate Karlodinium veneficum that is usually present at relatively low cell abundances is a globally-distributed harmful algal bloom-forming species, which negatively affects marine ecosystems, fisheries, and human health. Hence, an efficient detection platform for the rapid and sensitive identification of K. veneficum is highly demanded. In this study, a method referred to as recombinase polymerase amplification coupled with lateral flow dipstick (RPA-LFD) was developed for the rapid detection of K. veneficum. The primers for RPA and the detection probe for LFD were designed to specially target the internal transcribed spacer of K. veneficum by molecular cloning and multiple alignments of the related sequences. The developed RPA can gain an approximately 300 bp specific band from K. veneficum. Successful amplification for RPA could be achieved at a temperature range of 35 degrees C-45 degrees C. RPA for 30 min could produce enough products that could generate clearly visible electrophoresis bands and were adequate for subsequent LFD analysis. The RPA products can be visually detected by the naked eyes through an LFD after an automatic chromatography for 5 min at room temperature. The developed RPA-LFD was exclusively specific for K. veneficum and displayed no cross-reactivity with other algal species that are commonly distributed along the Chinese coast. In addition, the lowest detection limit of RPALFD was 10 ng mu L-1 of genomic DNA and 0.1 cell mL(-1), which was 100-fold sensitive than conventional PCR. In conclusion, the developed RPA-LFD assay in this study can be used as a rapid and sensitive method to monitor K. veneficum in the future. |
WOS研究方向 | Marine & Freshwater Biology |
来源期刊 | HARMFUL ALGAE |
文献类型 | 期刊论文 |
条目标识符 | http://gcip.llas.ac.cn/handle/2XKMVOVA/89816 |
作者单位 | 1.Harbin Inst Technol Weihai, Sch Marine Sci & Technol, Wenhua West Rd 2, Weihai 264209, Shandong, Peoples R China; 2.Ningbo Univ, Sch Marine Sci, Ningbo 315211, Zhejiang, Peoples R China; 3.Tsinghua Univ, Grad Sch Shenzhen, Div Ocean Sci & Technol, Shenzhen 518055, Peoples R China |
推荐引用方式 GB/T 7714 | Fu, Mengqi,Chen, Guofu,Zhang, Chunyun,et al. Rapid and sensitive detection method for Karlodinium veneficum by recombinase polymerase amplification coupled with lateral flow dipstick[J],2019,84:1-9. |
APA | Fu, Mengqi,Chen, Guofu,Zhang, Chunyun,Wang, Yuanyuan,Sun, Rui,&Zhou, Jin.(2019).Rapid and sensitive detection method for Karlodinium veneficum by recombinase polymerase amplification coupled with lateral flow dipstick.HARMFUL ALGAE,84,1-9. |
MLA | Fu, Mengqi,et al."Rapid and sensitive detection method for Karlodinium veneficum by recombinase polymerase amplification coupled with lateral flow dipstick".HARMFUL ALGAE 84(2019):1-9. |
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