Rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time PCR
Vibrio parahaemolyticus (V. parahaemolyticus) is acknowledged as one of the most significant disease-causing foodbome pathogen that causes the gastroenteritis resulting in diarrhea from vulnerable patients. The pathogenicity genes of V. parahaemolyticus are thermostable direct hemolysin (tdh) and th...
| Main Author: | |
|---|---|
| Format: | Final Year Project Report / IMRAD |
| Language: | English English |
| Published: |
Universiti Malaysia Sarawak, (UNIMAS)
2012
|
| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/8220/ http://ir.unimas.my/id/eprint/8220/1/Patricia.pdf http://ir.unimas.my/id/eprint/8220/4/Patricia%20Rowena.pdf |
| _version_ | 1848836327377731584 |
|---|---|
| author | Patricia Rowena, Mark Baran Saging |
| author_facet | Patricia Rowena, Mark Baran Saging |
| author_sort | Patricia Rowena, Mark Baran Saging |
| building | UNIMAS Institutional Repository |
| collection | Online Access |
| description | Vibrio parahaemolyticus (V. parahaemolyticus) is acknowledged as one of the most significant disease-causing foodbome pathogen that causes the gastroenteritis resulting in diarrhea from vulnerable patients. The pathogenicity genes of V. parahaemolyticus are thermostable direct hemolysin (tdh) and thermostable direct hemolysin related hemolysin (trh). However, targeting a gene of thermolabile hemolysin (tlh) gene in this study, is described in total V. parahaemolyticus which may play its role in affecting the host and contribute to human health risks. This research aims for V. parahemolyticus in rapidly detecting and quantitatively determines the present and the level of the bacteria from 30 samples of cockles (Anadara granosa) purchased from Kuching7'~-Mile wet market by utilizing the Real-Time PCR. In this study, the primer MV2B-tl (tlh-gene based) and the SYBR Green PCR mastermix were used in Real-Time PCR assay and among all the 30 samples of cockles isolated, there were no detectable quantification of V. parahaemolyticus. Quantification of V. parahaemolyticus could be determined by the standard curve result obtained. Spiking of 'Ti/apia' fish with Ixl07 CFU/mL V. parahaemolyticu aids in determining the sensitivity of the study by Real-Time PCR assay by standard curve plot obtained with ~ value of 0.99 which showed a good correlation. Furthermore, the specificity of the assay was confinned by testing with 7 non -Vibrio strains by specific PCR. The SYBR Green PCR mastermix and primer set were developed as quantification tool of V. parahaemolyticus to the traditional PCR method which needed post-peR result and is time consuming. Therefore, the development of Real-Time peR assay in analyzing the presence of bacteria in seafood is crucial to both seafood industries as well as to the consumers. |
| first_indexed | 2025-11-15T06:22:00Z |
| format | Final Year Project Report / IMRAD |
| id | unimas-8220 |
| institution | Universiti Malaysia Sarawak |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T06:22:00Z |
| publishDate | 2012 |
| publisher | Universiti Malaysia Sarawak, (UNIMAS) |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | unimas-82202024-02-26T07:54:46Z http://ir.unimas.my/id/eprint/8220/ Rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time PCR Patricia Rowena, Mark Baran Saging QL Zoology Vibrio parahaemolyticus (V. parahaemolyticus) is acknowledged as one of the most significant disease-causing foodbome pathogen that causes the gastroenteritis resulting in diarrhea from vulnerable patients. The pathogenicity genes of V. parahaemolyticus are thermostable direct hemolysin (tdh) and thermostable direct hemolysin related hemolysin (trh). However, targeting a gene of thermolabile hemolysin (tlh) gene in this study, is described in total V. parahaemolyticus which may play its role in affecting the host and contribute to human health risks. This research aims for V. parahemolyticus in rapidly detecting and quantitatively determines the present and the level of the bacteria from 30 samples of cockles (Anadara granosa) purchased from Kuching7'~-Mile wet market by utilizing the Real-Time PCR. In this study, the primer MV2B-tl (tlh-gene based) and the SYBR Green PCR mastermix were used in Real-Time PCR assay and among all the 30 samples of cockles isolated, there were no detectable quantification of V. parahaemolyticus. Quantification of V. parahaemolyticus could be determined by the standard curve result obtained. Spiking of 'Ti/apia' fish with Ixl07 CFU/mL V. parahaemolyticu aids in determining the sensitivity of the study by Real-Time PCR assay by standard curve plot obtained with ~ value of 0.99 which showed a good correlation. Furthermore, the specificity of the assay was confinned by testing with 7 non -Vibrio strains by specific PCR. The SYBR Green PCR mastermix and primer set were developed as quantification tool of V. parahaemolyticus to the traditional PCR method which needed post-peR result and is time consuming. Therefore, the development of Real-Time peR assay in analyzing the presence of bacteria in seafood is crucial to both seafood industries as well as to the consumers. Universiti Malaysia Sarawak, (UNIMAS) 2012 Final Year Project Report / IMRAD NonPeerReviewed text en http://ir.unimas.my/id/eprint/8220/1/Patricia.pdf text en http://ir.unimas.my/id/eprint/8220/4/Patricia%20Rowena.pdf Patricia Rowena, Mark Baran Saging (2012) Rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time PCR. [Final Year Project Report / IMRAD] (Unpublished) |
| spellingShingle | QL Zoology Patricia Rowena, Mark Baran Saging Rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time PCR |
| title | Rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time PCR |
| title_full | Rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time PCR |
| title_fullStr | Rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time PCR |
| title_full_unstemmed | Rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time PCR |
| title_short | Rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time PCR |
| title_sort | rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time pcr |
| topic | QL Zoology |
| url | http://ir.unimas.my/id/eprint/8220/ http://ir.unimas.my/id/eprint/8220/1/Patricia.pdf http://ir.unimas.my/id/eprint/8220/4/Patricia%20Rowena.pdf |