Saltatory rolling circle amplification (SRCA) combined with Safranine O for the rapid, sensitive and visual detection of Listeria monocytogenes in food
跨越式滚环等温扩增（SRCA）联合Safranine O快速、灵敏、直观检测食品中单核细胞增生李斯特氏菌
- 关键词：
- 来源：
- EUROPEAN FOOD RESEARCH AND TECHNOLOGY
- 类型：
- 学术文献
- 语种：
- 英语
- 原文发布日期：
- 2024-11-20
- 摘要：
- Listeria monocytogenes (L. monocytogenes) is one of the mostprevalent foodborne pathogens. It causes intestinal infections as well assevere meningitis and septicemia, representing a significant risk to foodsafety and human health. Consequently, the present study proposes a novelmethod based on the combination of saltatory rolling circle amplification(SRCA) technology and Safranine O for the rapid, sensitive and visual detectionof L. monocytogenes in food. In the presence of L.monocytogenes, the SRCA amplification reaction yields copiousquantities of double-stranded DNA. Following the addition of Safranine O andthe oxidant, Safranine O is oxidized to form Safranine O oligomer. Theelectrostatic interactions between the positively charged Safranine O oligomersand the negatively charged SRCA amplification product result in the formationof dark red visible aggregates, the absorbance value also decreases with theformation of aggregates. The method has low detection limit, high specificity,straightforward and time efficient (DNA amplification for 40 min, aggregationfor 10 min). The visual qualitative detection limit was 2.7 x 101 CFU/mL, and the limitof detection quantitative detection was 3.4 CFU/mL. In comparison with the SRCAelectrophoresis detection method, the visual detection limit was enhanced by 10times. Comparison with ISO method, the sensitivity, specificity and accuracy ofthe method for the detection of real samples were 100%, 96.2% and 96.7%,respectively. This method offers a straightforward and accurate qualitative andquantitative approach for the identification of L. monocytogenes in food, and isparticularly suited to the detection of pathogenic bacteria in food inresource-poor areas.
- 所属专题：
- 63
