표제지
목차
국문초록 12
I. 서론 14
II. 문헌 고찰 16
1. Listeria monocytogenes 16
1) 일반적 특성 16
2) 식중독 발병 현황 18
2. 세포막 용해버퍼 21
3. 분자학적 검출 방법 23
1) Conventional PCR 23
2) Real-time PCR 23
III. 재료 및 방법 25
1. 세포막 용해버퍼 구성성분 조사 25
2. 균액상에서 세포막 용해버퍼 적용 25
1) 세포막 용해버퍼 제조 25
2) 시험 균액 준비 27
3) Listeria monocytogenes의 최적 세포막 용해버퍼 선정 27
3. 식품에서 세포막 용해버퍼 평가 32
1) Listeria monocytogenes 검출 식품 조사 및 실험 대상 식품의 선정 32
2) 시료 준비 32
3) 시험 균액 준비 및 균액 접종 33
4) 세포막 용해버퍼 효과 확인 과정 33
4. 통계학적 분석 38
IV. 결과 및 고찰 41
1. 세포막 용해버퍼 구성성분 선정 41
2. 균액상에서 세포막 용해버퍼 효과 확인 44
가) Conventional PCR 44
나) 시간 및 비용 분석 61
3. 식품에서 세포막 용해버퍼의 효과 확인 63
1) Listeria monocytogenes 검출 식품 조사 및 실험 대상 식품의 선정 결과 63
2) 식품별 세포막 용해버퍼의 효과 확인 66
V. 요약 및 결론 75
VI. 참고문헌 78
ABSTRACT 86
Table 1. Growth conditions and survival limits of Listeria monocytogenes 17
Table 2. Outbreaks of Listeria monocytogenes infection in the United... 20
Table 3. List of surfactants used in cell lysis 22
Table 4. prfA gene primers designed for the detection of Listeria monocytogenes 29
Table 5. Conventional PCR conditions for identification of Listeria... 30
Table 6. hlyA gene primers designed for the detection of Listeria... 36
Table 7. Conventional PCR conditions for identification of Listeria... 37
Table 8. hlyA gene primers designed for the detection of Listeria... 39
Table 9. Real-time PCR conditions for identification of Listeria... 40
Table 10. Selection of membrane lysis buffer components 42
Table 11. Components of lysis buffer and cell lysis effect of Listeria monocytogenes 56
Table 12. Time and cost analysis according to cell membrane lysis buffers 62
Table 13. Detection of Listeria monocytogenes in foods 64
Table 14. Comparison of Ct values of Listeria monocytogenes in king oyster mushrooms through real-time PCR[이미지참조] 72
Table 15. Comparison of Ct values of Listeria monocytogenes in Jokbal through real-time PCR[이미지참조] 73
Table 16. Comparison of Ct values of Listeria monocytogenes in squids through real-time PCR[이미지참조] 74
Figure 1. Outbreaks of Listeria monocytogenes infection in Korea (2015-2020). 19
Figure 2. Combinations of lysis buffers. 26
Figure 3. Cell membrane lysis effect of distilled water, 1% SDS and additives... 45
Figure 4. Cell membrane lysis effect of distilled water, 0.05% digitonin and... 46
Figure 5. Cell membrane lysis effect of distilled water, 0.5% CHAPS and... 47
Figure 6. Cell membrane lysis effect of 1xPBS, 1% SDS and additives on... 48
Figure 7. Cell membrane lysis effect of 1xPBS, 0.05% digitonin and additives... 49
Figure 8. Cell membrane lysis effect of 1xPBS, 0.5% CHAPS and additives on... 50
Figure 9. Cell membrane lysis effect of 20 mM Tris-HCl, 1% SDS and additives on... 51
Figure 10. Cell membrane lysis effect of 20 mM Tris-HCl, 0.05% digitonin and... 52
Figure 11. Cell membrane lysis effect of 20 mM Tris-HCl, 0.5% CHAPS and... 53
Figure 12. First repeated experiments for the best cell membrane lysis buffer... 54
Figure 13. Second repeated experiments for the best cell membrane lysis buffer... 55
Figure 14. The low concentration of Listeria monocytogenes lysis effect on the... 59
Figure 15. Comparison of the detection effect of Listeria monocytogenes in king... 68
Figure 16. Comparison of the detection effect of Listeria monocytogenes in... 69
Figure 17. Comparison of the detection effect of Listeria monocytogenes in... 70