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Rapid detection of Pseudo-nitzschia species using whole-cell fluorescence in situ Hybridization (FISH)

The genus Pseudo-nitzschia comprised of thirty-four known species with half of them is reported to produce domaic acid (DA), a neurotoxin associated with Amnesic Shellfish Poisoning (ASP). Species identification of Pseudo-nitzschia mainly depends on detailed observations of the fine ultrastructure...

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Bibliographic Details
Main Author: Lim, Lay Peng
Format: Final Year Project Report
Language:English
Published: Universiti Malaysia Sarawak, (UNIMAS) 2011
Subjects:
Q Science (General)
QD Chemistry
SH Aquaculture. Fisheries. Angling
Online Access:http://ir.unimas.my/id/eprint/6434/8/Lim%20Lay%20Peng.pdf
http://ir.unimas.my/id/eprint/6434/
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spelling my.unimas.ir.64342023-12-11T08:58:57Z http://ir.unimas.my/id/eprint/6434/ Rapid detection of Pseudo-nitzschia species using whole-cell fluorescence in situ Hybridization (FISH) Lim, Lay Peng Q Science (General) QD Chemistry SH Aquaculture. Fisheries. Angling The genus Pseudo-nitzschia comprised of thirty-four known species with half of them is reported to produce domaic acid (DA), a neurotoxin associated with Amnesic Shellfish Poisoning (ASP). Species identification of Pseudo-nitzschia mainly depends on detailed observations of the fine ultrastructure of frustules which cannot be easily distinguishable without electron microscope. In this study, a molecular approach for rapid detection of Pseudo-nitzschia spp. by using whole-cell fluorescence in situ hybridization (FISH) was developed. Cultured and natural samples of Pseudo-nitzschia used in this study were identified by transmission and scanning electron microscopy (TEM & SEM). Species specific oligonucleotide probes were designed in silico to P. pungens (L-S-P.pu-0378-a-A-23), P. brasiliana (L-S-P.br-0129-a-A-23) and P. galaxiae (L-S-P.ga-0128-a-A-23) based on sequences obtained in this study and SILVA database. Synthesized P. pungens specie specific PuD1 probe was applied on both cultured and field samples with UniC and UniR probe used as positive and negative control. Efficiency of PuD1 probe in detection was consistently more than 93%. Probe PuD1 coupled with FISH method is equally efficiency in cell enumeration compared to traditional method with low discrepancy (<20%). This rapid detection method should be adopted in the HABs monitoring program for detection of harmful and potential harmful species of Pseudonitzschia. Universiti Malaysia Sarawak, (UNIMAS) 2011 Final Year Project Report NonPeerReviewed text en http://ir.unimas.my/id/eprint/6434/8/Lim%20Lay%20Peng.pdf Lim, Lay Peng (2011) Rapid detection of Pseudo-nitzschia species using whole-cell fluorescence in situ Hybridization (FISH). [Final Year Project Report] (Unpublished)
institution Universiti Malaysia Sarawak
building Centre for Academic Information Services (CAIS)
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Sarawak
content_source UNIMAS Institutional Repository
url_provider http://ir.unimas.my/
language English
topic Q Science (General)
QD Chemistry
SH Aquaculture. Fisheries. Angling
spellingShingle Q Science (General)
QD Chemistry
SH Aquaculture. Fisheries. Angling
Lim, Lay Peng
Rapid detection of Pseudo-nitzschia species using whole-cell fluorescence in situ Hybridization (FISH)
description The genus Pseudo-nitzschia comprised of thirty-four known species with half of them is reported to produce domaic acid (DA), a neurotoxin associated with Amnesic Shellfish Poisoning (ASP). Species identification of Pseudo-nitzschia mainly depends on detailed observations of the fine ultrastructure of frustules which cannot be easily distinguishable without electron microscope. In this study, a molecular approach for rapid detection of Pseudo-nitzschia spp. by using whole-cell fluorescence in situ hybridization (FISH) was developed. Cultured and natural samples of Pseudo-nitzschia used in this study were identified by transmission and scanning electron microscopy (TEM & SEM). Species specific oligonucleotide probes were designed in silico to P. pungens (L-S-P.pu-0378-a-A-23), P. brasiliana (L-S-P.br-0129-a-A-23) and P. galaxiae (L-S-P.ga-0128-a-A-23) based on sequences obtained in this study and SILVA database. Synthesized P. pungens specie specific PuD1 probe was applied on both cultured and field samples with UniC and UniR probe used as positive and negative control. Efficiency of PuD1 probe in detection was consistently more than 93%. Probe PuD1 coupled with FISH method is equally efficiency in cell enumeration compared to traditional method with low discrepancy (<20%). This rapid detection method should be adopted in the HABs monitoring program for detection of harmful and potential harmful species of Pseudonitzschia.
format Final Year Project Report
author Lim, Lay Peng
author_facet Lim, Lay Peng
author_sort Lim, Lay Peng
title Rapid detection of Pseudo-nitzschia species using whole-cell fluorescence in situ Hybridization (FISH)
title_short Rapid detection of Pseudo-nitzschia species using whole-cell fluorescence in situ Hybridization (FISH)
title_full Rapid detection of Pseudo-nitzschia species using whole-cell fluorescence in situ Hybridization (FISH)
title_fullStr Rapid detection of Pseudo-nitzschia species using whole-cell fluorescence in situ Hybridization (FISH)
title_full_unstemmed Rapid detection of Pseudo-nitzschia species using whole-cell fluorescence in situ Hybridization (FISH)
title_sort rapid detection of pseudo-nitzschia species using whole-cell fluorescence in situ hybridization (fish)
publisher Universiti Malaysia Sarawak, (UNIMAS)
publishDate 2011
url http://ir.unimas.my/id/eprint/6434/8/Lim%20Lay%20Peng.pdf
http://ir.unimas.my/id/eprint/6434/
_version_ 1787140463779643392
score 13.211869

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