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Combination of Enrichment and PCR in Rapid Semi-Quantification of Bacillus cereus in Fresh-Cut Vegetables

Authors
최유경Sujung Lee윤요한
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
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
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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