On-site rapid detection of toxic Alexandrium tamiyavanichii: integrating the species-specific hydrolysis probe in insulated isothermal polymerase chain reaction (iiPCR)
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 base...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Springer Netherlands
2016
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| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/11492/ http://ir.unimas.my/id/eprint/11492/1/On-site%20rapid%20detection%20of%20toxic%20Alexandrium%20tamiyavanichii%20%28abstract%29.pdf |
| Summary: | 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 |
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