Simultaneous differential detection of human pathogenic and nonpathogenic Vibrio species using a multiplex PCR based on gyrB and pntA genes
Aims: To develop a multiplex PCR targeting the gyrB and pntA genes for Vibrio species differentiation. Methods and Results: Four pairs of primers targeting gyrB gene of Vibrios at genus level and pntA gene of Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus were designed. This PCR method...
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Blackwell Publishing
2010
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| Online Access: | http://eprints.um.edu.my/3849/ |
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| _version_ | 1831446660211802112 |
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| author | Teh, C.S.J. Chua, K.H. Thong, Kwai Lin |
| author_facet | Teh, C.S.J. Chua, K.H. Thong, Kwai Lin |
| author_sort | Teh, C.S.J. |
| building | UM Library |
| collection | Institutional Repository |
| content_provider | Universiti Malaya |
| content_source | UM Research Repository |
| continent | Asia |
| country | Malaysia |
| description | Aims: To develop a multiplex PCR targeting the gyrB and pntA genes for Vibrio species differentiation. Methods and Results: Four pairs of primers targeting gyrB gene of Vibrios at genus level and pntA gene of Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus were designed. This PCR method precisely identified 250 Vibrio species and demonstrated sensitivity in the range of 4 -104 CFU ml-1 (c. 200 CFU per PCR) to 2 -103 CFU ml-1 (c. 10 CFU per PCR). Overall, the gyrB gene marker showed a higher specificity than the dnaJ gene marker for Vibrio detection and was able to distinguish Aeromonas from Vibrio species. Conclusions: The multiplex PCR based on combined gyrB and pntA provides a high discriminatory power in the differentiation between Vibrio alginolyticus and V. parahaemolyticus, and between V. cholerae and Vibrio mimicus. Significance and Impact of the Study: This assay will be useful for rapid differentiation of various Vibrio species from clinical and environmental sources and significantly overcomes the limitations of the conventional methods. |
| format | Article |
| id | my.um.eprints-3849 |
| institution | Universiti Malaya |
| publishDate | 2010 |
| publisher | Blackwell Publishing |
| record_format | eprints |
| spelling | my.um.eprints-38492018-10-15T04:18:58Z http://eprints.um.edu.my/3849/ Simultaneous differential detection of human pathogenic and nonpathogenic Vibrio species using a multiplex PCR based on gyrB and pntA genes Teh, C.S.J. Chua, K.H. Thong, Kwai Lin R Medicine Aims: To develop a multiplex PCR targeting the gyrB and pntA genes for Vibrio species differentiation. Methods and Results: Four pairs of primers targeting gyrB gene of Vibrios at genus level and pntA gene of Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus were designed. This PCR method precisely identified 250 Vibrio species and demonstrated sensitivity in the range of 4 -104 CFU ml-1 (c. 200 CFU per PCR) to 2 -103 CFU ml-1 (c. 10 CFU per PCR). Overall, the gyrB gene marker showed a higher specificity than the dnaJ gene marker for Vibrio detection and was able to distinguish Aeromonas from Vibrio species. Conclusions: The multiplex PCR based on combined gyrB and pntA provides a high discriminatory power in the differentiation between Vibrio alginolyticus and V. parahaemolyticus, and between V. cholerae and Vibrio mimicus. Significance and Impact of the Study: This assay will be useful for rapid differentiation of various Vibrio species from clinical and environmental sources and significantly overcomes the limitations of the conventional methods. Blackwell Publishing 2010 Article PeerReviewed Teh, C.S.J. and Chua, K.H. and Thong, Kwai Lin (2010) Simultaneous differential detection of human pathogenic and nonpathogenic Vibrio species using a multiplex PCR based on gyrB and pntA genes. Journal of Applied Microbiology, 108 (6). pp. 1940-1945. ISSN 1364-5072, |
| spellingShingle | R Medicine Teh, C.S.J. Chua, K.H. Thong, Kwai Lin Simultaneous differential detection of human pathogenic and nonpathogenic Vibrio species using a multiplex PCR based on gyrB and pntA genes |
| title | Simultaneous differential detection of human pathogenic and nonpathogenic Vibrio species using a multiplex PCR based on gyrB and pntA genes |
| title_full | Simultaneous differential detection of human pathogenic and nonpathogenic Vibrio species using a multiplex PCR based on gyrB and pntA genes |
| title_fullStr | Simultaneous differential detection of human pathogenic and nonpathogenic Vibrio species using a multiplex PCR based on gyrB and pntA genes |
| title_full_unstemmed | Simultaneous differential detection of human pathogenic and nonpathogenic Vibrio species using a multiplex PCR based on gyrB and pntA genes |
| title_short | Simultaneous differential detection of human pathogenic and nonpathogenic Vibrio species using a multiplex PCR based on gyrB and pntA genes |
| title_sort | simultaneous differential detection of human pathogenic and nonpathogenic vibrio species using a multiplex pcr based on gyrb and pnta genes |
| topic | R Medicine |
| url | http://eprints.um.edu.my/3849/ |
| url_provider | http://eprints.um.edu.my/ |