지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2011.1
- 수록면
- 158 - 165 (8page)
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A sensitive reverse transcriptase-PCR (RT-PCR) method to detect viable Escherichia coli O157:H7 in milk was established. The primer sets were designed based on the nucleotide sequences of the rfbE (per) and wbdN genes in the O157 antigen gene cluster of E. coli O157:H7. RT-PCR using five different primer sets yielded DNA with sizes of 655, 518, 450, and 149-bp, respectively. All five of the E. coli O157:H7 strains were detected by RT-PCR, but 11 other bacterial species were not. The sensitivity of RT-PCR was improved by adding yeast tRNA as a carrier to the crude RNA extract. The RT-PCR amplifying the 149-bp DNA fragment was the most sensitive for detecting E. coli O157:H7 and the most refractory to the bactericidal treatments. Heat treatment at $65^{\circ}C$ for 30 min was the least inhibitory of all bactericidal treatments. Treatment with RNase A strongly inhibited the RT-PCR of heated milk but not unheated milk. This study described RT-PCR methods that are specific and sensitive with a detection limit of 10 E. coli O157:H7 cells, and showed that pre-treating milk samples with RNase A improved the specificity to detect viable bacteria by RT-PCR.
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참고문헌 (29)
참고문헌 신청
Bickley, J. / 1996 / Polymerase chain reaction (PCR) detection of Listeriamonocytogenes in diluted milk and reversal of PCR inhibitioncaused by calcium ions / Lett. Appl. Microbiol 22:153~158
Bilge, S. S. / 1996 / Role of the Escherichia coli O157:H7 O side chain in adherenceand analysis of rfb locus / Infect. Immun 64:4795~4801
de Wet, S. S. / 2008 / An improved method for RNA extraction from carcasssamples for detection of viable Escherchia coli O157:H7 byreverse-transcriptase polymerase chain reaction / Lett. Appl.Microbiol 47:399~404
Dineen, S. / 1998 / Persistence of Escherichia coli O157:H7 in dairy fermentationsystems / J. Food Prot 61:1602~1608
Doyle, M. T. / 1997 / Escherichiacoli O157:H7 / ASM Press :171~191
Bilge, S. S. / 1996 / Role of the Escherichia coli O157:H7 O side chain in adherenceand analysis of rfb locus / Infect. Immun 64:4795~4801
de Wet, S. S. / 2008 / An improved method for RNA extraction from carcasssamples for detection of viable Escherchia coli O157:H7 byreverse-transcriptase polymerase chain reaction / Lett. Appl.Microbiol 47:399~404
Dineen, S. / 1998 / Persistence of Escherichia coli O157:H7 in dairy fermentationsystems / J. Food Prot 61:1602~1608
Doyle, M. T. / 1997 / Escherichiacoli O157:H7 / ASM Press :171~191
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