On-site rapid detection of toxic Alexandrium tamiyavanichii: integrating the species-specific hydrolysis probe in insulated isothermal polymerase chain reaction (iiPCR)

Nyuk, Fong Kon and Lik, Winne Sing Lau and Ing, Kuo Law and Po, Teen Lim and Chui, Pin Leaw (2016) On-site rapid detection of toxic Alexandrium tamiyavanichii: integrating the species-specific hydrolysis probe in insulated isothermal polymerase chain reaction (iiPCR). Journal of Applied Phycology, 2016. pp. 1-6. ISSN 1573-5176

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Abstract

On-site investigation of phytoplankton samples is important for rapid detection of harmful algal species and for early warning of harmful algal bloom. Molecular detection method by DNA amplification in a portable insulated isothermal PCR (iiPCR) device provides a simple and rapid detection based on fluorescent probe within an hour of reaction time. The assay was developed for a paralytic shellfish toxinproducing dinoflagellate Alexandrium tamiyavanichii. The assay presents the data as positive or negative on the presence or absence of A. tamiyavanichii cells, with a limit of detection (LOD) at five target cells per reaction. While the assay is incapable to accurately quantify cell density, it exhibits high detection accuracy and strongly correlated with quantitative PCR (qPCR) data. The user repeatability of iiPCR assay was evaluated; the results showed that no significant differences in the assay run by different operators. Field applicability of the assay was further validated by environmental samples. Despite the shortcoming of the assay, the overall performance of the assay to detect cells, its low-cost effectiveness, and portability for on-site detection, iiPCR has proven its potential as an early screening tool for harmful algae monitoring. Nyuk Fong Kon1 & Winnie Lik Sing Lau2 & Ing Kuo Law2 & Po Teen Lim2 & Chui Pin Leaw2

Item Type: Article
Uncontrolled Keywords: Alexandrium, Dinoflagellate, Insulated isothermal polymerase chain reaction (iiPCR), Internal transcribed spacer (ITS), Paralytic shellfish poisoning, Rapid molecular detection, unimas, university, universiti, Borneo, Malaysia, Sarawak, Kuching, Samarahan, ipta, education, research, Universiti Malaysia Sarawak
Subjects: G Geography. Anthropology. Recreation > GE Environmental Sciences
Divisions: Academic Faculties, Institutes and Centres > Institute of Biodiversity and Environmental Conservation
Depositing User: Karen Kornalius
Date Deposited: 19 Apr 2016 00:43
Last Modified: 21 Oct 2016 01:51
URI: http://ir.unimas.my/id/eprint/11492

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