표제지
목차
Ⅰ. 서론 8
Ⅱ. 재료 및 방법 11
1. 공시균주 및 성장조건 11
2. 김밥 시료 준비 및 총생균수 측정 12
3. 공시균주 및 김밥 시료로부터의 총 DNA 추출 13
4. Multiplex PCR 반응 13
5. Agarose gel 전기영동 15
Ⅲ. 결과 및 고찰 17
1. Multiplex PCR법에서의 primer의 특이성 17
2. Multiplex PCR법에서의 primer의 민감성 19
3. 총생균수 확인 21
4. Total DNA 확인 23
5. 김밥에서 식품위해세균 검출에 대한 multiplex PCR의 적용 25
6. multiplex PCR에 대한 종합평가 30
Ⅳ. 적요 33
참고문헌 36
Abstract 42
Table 1. List of bacterial strains and medium used in the study 11
Table 2. List of oligonucleotide primers for multiplex PCR 14
Table 3. PCR condition of multiplex 15
Table 4. PCR condition of uniplex 15
Table 5. Distribution of the total visible cell numbers in the 30 kimbab samples at different sample periods (June, July, and August) 22
Fig. 1. Agarose gel electrophoresis of the uniplex PCR products obtained for food-borne pathogenic bacteria. Lane M; Size marker, 100 bp DNA ladder(Bioneer Inc., Seoul, Korea), lane 1; S. aureus(KCTC 1916), lane 2; S. enterica(KCTC 12456), lane 3; S. enteritids(KCTC 12400),... 18
Fig. 2. Agarose gel electrophoresis of the multiplex PCR products obtained For food-borne pathogenic bacteria. Lane M; Size marker, 100 bp DNA ladder(Bioneer Inc., Seoul, Korea), lane 1; 0 hr incubation of 0.32 log10 CFU/ml, lane 2; 4 hr incubation of 3.87 log10 CFU/ml,... 20
Fig. 3. Agarose gel electrophoresis of the total DNA obtained for kimbab samples. Lane M; Size marker, λ/PstI(Bioneer Inc., Seoul, Korea), lane 01 to 10; kimbab sample. 23
Fig. 4. Agarose gel electrophoresis of the total DNA obtained for kimbab samples. Lane M; Size marker, λ/PstI(Bioneer Inc., Seoul, Korea), lane 11 to 20; kimbab sample. 24
Fig. 5. Agarose gel electrophoresis of the total DNA obtained for kimbab samples. Lane M; Size marker, λ/PstI(Bioneer Inc., Seoul, Korea), lane 21~30; kimbab sample. 24
Fig. 6. Agarose gel electrophoresis of the multiplex PCR products obtained for kimbab samples. Lane M; Size marker, 100 bp DNA ladder(Bioneer Inc., Seoul, Korea), DNA ladder(Bioneer Inc., Seoul, Korea), lane 01; KB01 (B. cereus, S. aureus), lane 02; KB02 (S. aureus), lane... 28
Fig. 7. Agarose gel electrophoresis of the multiplex PCR products obtained for kimbab samples. Lane M; Size marker, 100 bp DNA ladder(Bioneer Inc., Seoul, Korea), lane 11; KB11 (B. cereus, Salmonella spp., S. aureus), lane 12; KB12 (Not detected), lane 13; KB13 (B. cereus),... 29
Fig. 8. Agarose gel electrophoresis of the multiplex PCR products obtained for kimbab samples. Lane M; Size marker, 100 bp DNA ladder(Bioneer Inc., Seoul, Korea), lane 21; KB21 (B, cereus, Salmonella spp., S, aureus), lane 22; KB22 (B, cereus, S, aureus), lane 23;... 30
Fig. 9. The contamination rate of food-borne pathogenic bacteria in 30 kimbab samples. 32
Fig. 10. Distribution of the food-borne pathogenic bacteria in each of sampling periods such as, June, July, and August. Numbers in square brackets give the total number of the kimbab samples in that sample periods. 33