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Multiplex PCR amplification of 16S rRNA gene¡BvirA¡Btpl¡Band H1d genes was developed enabling simultaneous detection in Escherichia coli¡Aan indicator of fecal contamination and food-borne microbial pathogens¡AShigella flexneri¡BCitrobacter freundii¡BSalmonella typhi¡BVibrio cholerae¡BVibrio parahaemolyticus¡Band Staphylococcus aureus¡CEach of the nine pairs of oligonucleotide primers was found to support PCR amplifications of only its targeted gene¡CThe optimized multiplex PCR reaction utilized a primer annealing temperature of 59 ¢Jand used agarose gel electrophoresis for detection of the PCR-amplified products¡CSelection of appropriate target genes¡Boligonucleotide primers ¡BPCR reaction¡Band cycling parameters resulted in the amplification of four target genes simultaneously in a single PCR reaction with the sensitivity of detection was 102 CFU after 32 cycles¡CMultiplex PCR amplification followed by differential PCR for E. coli / Shigella¡A and Citrobacter / Salmonella¡Asequenced for the PCR-amplified products of 16S rRNA gene of the seven pathogens in this study¡Aand used restriction endonuclease AfaI to confirm the PCR-amplified products of V. cholerae¡AV. parahaemolyticus and Staphylococcus aureus¡Ahas been shown to be an sensitive¡Aspecific¡Aand rapid method to detect food-borne bacterial pathogens¡C
| Identifer | oai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0914101-113959 |
| Date | 14 September 2001 |
| Creators | Hwang, Chung-Hsing |
| Contributors | Chung-Lung Cho, David Chao, Zheng-tze Cheng |
| Publisher | NSYSU |
| Source Sets | NSYSU Electronic Thesis and Dissertation Archive |
| Language | Cholon |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0914101-113959 |
| Rights | unrestricted, Copyright information available at source archive |
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