| Autor: |
Apirak, Visetsripong, Kobchai, Pattaragulwanit, Jiraporn, Thaniyavarn, Ryosuke, Matsuura, Akio, Kuroda, Orasa, Sutheinkul |
| Rok vydání: |
2007 |
| Předmět: |
|
| Zdroj: |
The Southeast Asian journal of tropical medicine and public health. 38(1) |
| ISSN: |
0125-1562 |
| Popis: |
A rapid method for detection of Escherichia coli O157: H7 using multiplex PCR was developed. Two oligonucleotide primer pairs were used for simultaneously detection of vt encoding verotoxin genes for virulence factor and rfb(O157) encoding the O-antigen specific for E. coli O157: H7. Multiplex PCR generated two products of 215 bp and 420 bp for vt and rfb(O157), respectively. Multiplex PCR detected reference strain O157: H7 (NF-7777) with a sensitivity of 10(5) CFU per ml with no amplification of other 15 pathogenic bacteria. After incubation of 10(2) CFU/25 gram raw meat in tryptic soy broth at 37 degrees C for 8 hours, multiplex PCR conducted with the addition of 100 mg bovine serum albumin produced the two specific PCR products for E. coli O157: H7. This modified multiplex PCR is a rapid, sensitive, and specific technique for detecting and differentiating E. coli O157: H7 and has the potential to be used as an alternative to conventional methods for the screening of O157: H7 strains isolated from raw meat. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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