(食品质量和 与安全研讨会)he study of multiplex PCR-based rapid detection technique for foodborne pathogen on fresh-cut fruits and vegetables.pptVIP

Table. 2. The colony count of L. monocytogenes, E. coli O157:H7 and S. aureus inoculated on fresh-cut fruit based dual filtration membrane The filtration of pathogen Filtration (log cfu/ml) Polypropylene membrane (log cfu/ml) Nylon membrane (log cfu/mll) 10μM 20μM 40μM 15μM 40μM 60μM Re-filtration 0.22 0.45 0.22 0.45 0.22 0.45 0.22 0.45 0.22 0.45 0.22 0.45 L. monocytogenes 8.90 7.20 7.08 7.45 7.20 7.81 7.63 8.58 8.18 8.60 8.48 8.63 8.11 E. coli O157:H7 8.70 7.57 7.18 7.71 7.49 7.85 7.62 8.15 7.97 8.23 7.93 8.34 8.18 S. aureus 8.38 7.69 7.38 7.78 7.54 7.98 7.73 8.04 7.94 8.08 8.04 8.23 7.99 Different types of filtration were chosen to enhance detection limitation and save detection time. Fig. 8. The LOD of multiplex PCR assay using 10-fold serially diluted the population of L. monocytogenes, E. coli O157:H7 and S. aureus on fresh-cut fruits. M: 1000bp DNA marker. Lane 1- Lane 8 showed amplicon result for L. monocytogenes, E. coli O157:H7 and S. aureus (from 108 cfu/ml to 10 cfu/ml). Lane 9 and Lane 10 was negative control. The detection limitation of pathogen on fresh-cut fruit Fig.5 The result showed detection limitation for three pathogen was 104 cfu/ml, while for L. monocytogenes was 104 cfu/ml, for E. coli O157:H7 was 102 cfu/ml, S. aureus was 103 cfu/ml. Conclusion 1. The newly designed filtration-base PMA-mPCR assay in this paper was sensitive, specific and rapid for simultaneous detection of viable L. monocytogenes, S. aureus and E. coli O157:H7 on fresh-cut fruit. 2. Combination of the PMA-mPCR assay with filtration membrane could eliminate the effect of inhibitor from food sample effectively. 3. This study showed the method was considered to be more successful in detecting L. monocytogenes, S. aureus and E.coli O157:H7. Conclusions ◎已发表论文： 1. 冯 可，胡文忠，姜爱丽，徐永平，萨仁高娃，预测微生物学在鲜切果蔬产品质量安全控制中的应用，食品工业 科技， 2014，35（10）：49-52，56 2. 冯 可，胡文忠，姜爱丽，徐永平，萨仁高娃. 单核细胞增生性李斯特菌分子生物学检测技术的研究进展，食品工 业科技. 2014，35（4）：392-396 3. 萨仁高娃，胡文忠，姜爱丽，马杰，冯可，产单核细胞增生性李斯特氏菌致病机制