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Rapid Detection of Listeria monocytogenes by Real-Time PCR in Processed Meat and Dairy Products

Authors
Heo, Eun JeongSong, Bo RaPark, Hyun JungKim, Young JoMoon, Jin SanWee, Sung HwanKim, Jin-SeokYoon, Yohan
Issue Date
Mar-2014
Publisher
INT ASSOC FOOD PROTECTION
Citation
JOURNAL OF FOOD PROTECTION, v.77, no.3, pp 453 - 458
Pages
6
Journal Title
JOURNAL OF FOOD PROTECTION
Volume
77
Number
3
Start Page
453
End Page
458
URI
https://scholarworks.sookmyung.ac.kr/handle/2020.sw.sookmyung/10967
DOI
10.4315/0362-028X.JFP-13-318
ISSN
0362-028X
1944-9097
Abstract
The objectives of this study were to evaluate the detection of Listeria monocyto genes in different ready-to-eat foods using real-time PCR (RT-PCR). Various concentrations (10(0) to 10(5) CFU/ml) of L. monocytogenes ATCC 19115 were inoculated into ham, sausage, ground meat, processed milk, cheese, and infant formula. L. monocytogenes ATCC 19115 in the samples was then enumerated on Oxford agar, and DNA was extracted from the samples before and after incubation at 36 degrees C for 4 h. A set of primers and hybridization probe designed in this study was then used to detect the pathogen. The standard curve was then prepared by plotting cycle threshold values for each dilution versus L. monocyto genes cell counts (log CPU). The specificity of the set of primers and hybridization probe was appropriate. A 4-h incubation at 36 degrees C before DNA extraction produced optimum standard curves in comparison to the results for a 0-h incubation. Thus, a 4-h incubation at 36 degrees C was applied for monitoring L. monocytogenes in collected food samples. To monitor L. monocyto genes in foods, 533 samples (ham, 129; sausage, 226; ground meat, 72; processed cheese, 54; processed milk, 42; and infant formula, 10) were collected from retail markets and from the step before pasteurization in plants. Of all 533 samples, 4 samples (0.8%) showed positive signals in RT-PCR. Two samples from hams (1.6%) and two samples from sausages (0.9%) were determined to be positive for L. monocytogenes at <100 CFU/g. The results indicate that the RT-PCR detection method with the set of primers and hybridization probe designed in this study should be useful in monitoring for L. monocytogenes in processed meat and milk products.
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