Combination of Enrichment and PCR in Rapid Semi-Quantification of Bacillus cereus in Fresh-Cut Vegetables
- Authors
- 최유경; Sujung Lee; 윤요한
- Issue Date
- Aug-2020
- Publisher
- 한국식품위생안전성학회
- Keywords
- Bacillus cereus; Fresh-cut vegetables; Rapid detection; Enrichment; PCR
- Citation
- 한국식품위생안전성학회지, v.35, no.4, pp 319 - 325
- Pages
- 7
- Journal Title
- 한국식품위생안전성학회지
- Volume
- 35
- Number
- 4
- Start Page
- 319
- End Page
- 325
- URI
- https://scholarworks.sookmyung.ac.kr/handle/2020.sw.sookmyung/1263
- DOI
- 10.13103/JFHS.2020.35.4.319
- ISSN
- 1229-1153
2465-9223
- Abstract
- Widespread consumption of fresh-cut vegetables without cooking results in ingestion of major foodborne pathogens including Bacillus cereus. In this study, we aimed to develop a method to rapidly detect B. cereus in fresh-cut vegetables by combining commercial PCR analysis with enrichment of the pathogenic levels. A mixture of B. cereus strains (KCTC1013, KCTC1014, KCTC1092, KCTC1094, and KCTC3624) was inoculated on the surface of fresh-cut cabbage lettuce (20 g) and baby leafy vegetables (10 g) to concentration 1, 2, 3, 4, and 5 log CFU/g. Eighty milliliters of TSB with 0.15% polymyxin B was used for cabbage lettuce, and 90 mL of medium was used for baby leafy vegetables and incubated at 42 o C for 0, 2, 3, 4, 5, 6, and 7 h. One milliliter of the enriched media was plated on mannitol-egg yolk-polymyxin agar for quantification, and another 1 mL was used for DNA extraction for PCR analysis. Additionally, the minimum number of sub-samples to be tested from a pack of fresh-cut vegetable samples was determined using 5 sub-samples. The results from this study showed that for detecting B. cereus in fresh-cut cabbage lettuce, 3, 4, 5, 6, and 7 h enrichment were required to at least detect 5, 4, 3, 2, and 1 log CFU/g of B. cereus, respectively. B. cereus in fresh-cut baby leafy vegetables could be detected after 2, 3, 4, 5, and 6 h of enrichment at 5, 4, 3, 2, and 1 log CFU/g, respectively, using a combination of enrichment and PCR analysis. To determine if a pack of fresh-cut vegetable is positive, the minimum number of sub-samples should be 3. These results can be used to develop a rapid detection method to semi-quantify B. cereus in fresh-cut vegetable samples combining enrichment and PCR.
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