Application of whole-cell fluorescent in situ hybridization (FISH) of molecular detection of toxic alexandrium species
Paralytic shellfish poisoning is a type of seafood poisoning due to contamination of a group of neurotoxin, collectively known as saxitoxin, in filter feeding mollusks. Monitoring of PSP producing organism especially from the genus of Alexandrium is very challenging as identification of toxic spec...
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| Format: | Final Year Project Report / IMRAD |
| Language: | English |
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Universiti Malaysia Sarawak, (UNIMAS)
2011
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| Online Access: | http://ir.unimas.my/id/eprint/6403/ http://ir.unimas.my/id/eprint/6403/8/APPLICATION%20OF%20WHOLE-CELL%20FLUORESCENT%20IN%20SITU%20HYBRIDIZATION%20%28FISH%29%28OCR%29.pdf |
| _version_ | 1848835909816942592 |
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| author | Cheah, Bi Xuan |
| author_facet | Cheah, Bi Xuan |
| author_sort | Cheah, Bi Xuan |
| building | UNIMAS Institutional Repository |
| collection | Online Access |
| description | Paralytic shellfish poisoning is a type of seafood poisoning due to contamination of a group of neurotoxin,
collectively known as saxitoxin, in filter feeding mollusks. Monitoring of PSP producing organism especially
from the genus of Alexandrium is very challenging as identification of toxic species is only possible based on
plate tabulation. In this study, oligonucleotide probes using whole cell fluorescence in situ hybridization
(FISH) to detect toxic A. tamiyavanichii and A. minutum were developed. In silico probe design for the 2
species was carried out using sequences database of Alexandrium species found in Malaysia. Probe specificity was assessed based on GC content, melting temperature, Gibbs free energy change and E value. Potential species-specific probes for A. minutum and A. tamiyavanichii were designated as [L-S-Amin-680- (A. minutum)-A-24] and [L-S-Atam-286-(A. tamiyavanichii)-A-23]. Specific probe for A. tamiyavanichii, TAMIS1 was synthesized and tested against A. tamiyavanichii and all other related species. Probe uniC and uniR were applied as positive and negative control. Our result show high specificity of TAMIS1 to A.
tamiyavanichii with no cross-reactivity to four other Alexandrium (A. minutum, A. leei, A. tamarense and
A.affine). FISH condition applied in this study with hybridization temperature, 35 °C and formamide 40 % work well with no unspecific binding observed. The probe TAMIS1 should be adopted in national HAB monitoring program in order to increase the efficiency in detection of A. tamiyavanichii. |
| first_indexed | 2025-11-15T06:15:22Z |
| format | Final Year Project Report / IMRAD |
| id | unimas-6403 |
| institution | Universiti Malaysia Sarawak |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T06:15:22Z |
| publishDate | 2011 |
| publisher | Universiti Malaysia Sarawak, (UNIMAS) |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | unimas-64032023-02-06T06:50:36Z http://ir.unimas.my/id/eprint/6403/ Application of whole-cell fluorescent in situ hybridization (FISH) of molecular detection of toxic alexandrium species Cheah, Bi Xuan Q Science (General) SH Aquaculture. Fisheries. Angling Paralytic shellfish poisoning is a type of seafood poisoning due to contamination of a group of neurotoxin, collectively known as saxitoxin, in filter feeding mollusks. Monitoring of PSP producing organism especially from the genus of Alexandrium is very challenging as identification of toxic species is only possible based on plate tabulation. In this study, oligonucleotide probes using whole cell fluorescence in situ hybridization (FISH) to detect toxic A. tamiyavanichii and A. minutum were developed. In silico probe design for the 2 species was carried out using sequences database of Alexandrium species found in Malaysia. Probe specificity was assessed based on GC content, melting temperature, Gibbs free energy change and E value. Potential species-specific probes for A. minutum and A. tamiyavanichii were designated as [L-S-Amin-680- (A. minutum)-A-24] and [L-S-Atam-286-(A. tamiyavanichii)-A-23]. Specific probe for A. tamiyavanichii, TAMIS1 was synthesized and tested against A. tamiyavanichii and all other related species. Probe uniC and uniR were applied as positive and negative control. Our result show high specificity of TAMIS1 to A. tamiyavanichii with no cross-reactivity to four other Alexandrium (A. minutum, A. leei, A. tamarense and A.affine). FISH condition applied in this study with hybridization temperature, 35 °C and formamide 40 % work well with no unspecific binding observed. The probe TAMIS1 should be adopted in national HAB monitoring program in order to increase the efficiency in detection of A. tamiyavanichii. Universiti Malaysia Sarawak, (UNIMAS) 2011 Final Year Project Report / IMRAD NonPeerReviewed text en http://ir.unimas.my/id/eprint/6403/8/APPLICATION%20OF%20WHOLE-CELL%20FLUORESCENT%20IN%20SITU%20HYBRIDIZATION%20%28FISH%29%28OCR%29.pdf Cheah, Bi Xuan (2011) Application of whole-cell fluorescent in situ hybridization (FISH) of molecular detection of toxic alexandrium species. [Final Year Project Report / IMRAD] (Unpublished) |
| spellingShingle | Q Science (General) SH Aquaculture. Fisheries. Angling Cheah, Bi Xuan Application of whole-cell fluorescent in situ hybridization (FISH) of molecular detection of toxic alexandrium species |
| title | Application of whole-cell fluorescent in situ hybridization (FISH) of molecular detection of toxic alexandrium species |
| title_full | Application of whole-cell fluorescent in situ hybridization (FISH) of molecular detection of toxic alexandrium species |
| title_fullStr | Application of whole-cell fluorescent in situ hybridization (FISH) of molecular detection of toxic alexandrium species |
| title_full_unstemmed | Application of whole-cell fluorescent in situ hybridization (FISH) of molecular detection of toxic alexandrium species |
| title_short | Application of whole-cell fluorescent in situ hybridization (FISH) of molecular detection of toxic alexandrium species |
| title_sort | application of whole-cell fluorescent in situ hybridization (fish) of molecular detection of toxic alexandrium species |
| topic | Q Science (General) SH Aquaculture. Fisheries. Angling |
| url | http://ir.unimas.my/id/eprint/6403/ http://ir.unimas.my/id/eprint/6403/8/APPLICATION%20OF%20WHOLE-CELL%20FLUORESCENT%20IN%20SITU%20HYBRIDIZATION%20%28FISH%29%28OCR%29.pdf |