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Publication
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| Research Title |
DEVELOPMENT OF MULTIPLEX PCR AND REAL-TIME SYBR GREEN PCR FOR DETECTION OF FIVE-ENTEROPATHOGENIC BACTERIA IN RECTAL SWABS |
| Date of Distribution |
4 October 2012 |
| Conference |
| Title of the Conference |
INTERNATIONAL CONFERENCE ON MICROBIAL TAXONOMY, BASIC AND APPLIED MICROBIOLOGY |
| Organiser |
Department of Microbiology, Faculty of Science, Khon Kaen University, Thailand |
| Conference Place |
Kosa Hotel |
| Province/State |
Khon Kaen, Thailand |
| Conference Date |
4 October 2012 |
| To |
6 October 2012 |
| Proceeding Paper |
| Volume |
- |
| Issue |
- |
| Page |
- |
| Editors/edition/publisher |
- |
| Abstract |
Salmonella spp., Shigella spp., Vibrio cholerae, Vibrio parahaemolyticus and Staphylococcus aureus are the important agents caused diarrheal diseases. In this study, we developed multiplex PCR and two sets of duplex real-time SYBR Green PCR (dRT-SYBR) for detection of five-pathogens in rectal swab with and without enrichment. Total of 153 rectal swab samples were investigated for five-pathogens by those assays compared with conventional culture method. The sensitivity of multiplex PCR for the simultaneous detection of five-pathogens in mixed pure cultures was 103 CFU/ml (102 CFU/PCR reaction). After enrichment 6 h, the limit of detection (LOD) of the artificially inoculated rectal swabs was 10 CFU/ml (1 CFU/PCR reaction). After enrichment 3 h, the two sets of duplex real-time SYBR Green PCR (dRT-SYBR) for co-amplification of S. aureus and V. parahaemolyticus and another set for Salmonella spp. and V. cholerae showed the same LOD as 10 CFU/ml (1 CFU/PCR reaction). Some bacterial pathogens found in rectal swabs were 25.5% (39/153). Of 153 rectal swab samples, 16.3% (25/153) were detected by culture method, 25.5% (39/153) by multiplex PCR and 25.5% (39/153) by RT-SYBR PCR assay. Conclusion: The developed multiplex PCR assay and duplex real-time SYBR Green PCR are sensitive and effective methods for simultaneous detection of enteropathogenic bacteria in rectal swab specimens and serve as diagnostic tools for diarrheal patients. |
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| Peer Review Status |
ไม่มีผู้ประเมินอิสระ |
| Level of Conference |
นานาชาติ |
| Type of Proceeding |
Abstract |
| Type of Presentation |
Oral |
| Part of thesis |
true |
| Presentation awarding |
false |
| Attach file |
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| Citation |
0
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Journal Publication
ไทย