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Development and application of a new multiplex real‐time PCR assay for simultaneous identification of Brucella melitensis , Cronobacter sakazakii and Listeria monocytogenes in raw milk and cheese
Author(s) -
Tutar Esen,
Akıncı Kübra Sueda,
Akyol İsmaİl
Publication year - 2018
Publication title -
international journal of dairy technology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.061
H-Index - 53
eISSN - 1471-0307
pISSN - 1364-727X
DOI - 10.1111/1471-0307.12500
Subject(s) - brucella melitensis , listeria monocytogenes , raw milk , cronobacter sakazakii , multiplex polymerase chain reaction , multiplex , microbiology and biotechnology , brucella , biology , food safety , food science , brucellosis , bacteria , polymerase chain reaction , virology , gene , infant formula , genetics , bioinformatics , biochemistry
Brucella melitensis , Cronobacter sakazakii and Listeria monocytogenes are important foodborne pathogens in milk and milk products, which are responsible for a variety of diseases that pose serious hazards to public health and food safety. The objective of this study was to develop a novel multiplex RT i‐ PCR for the detection of B. melitensis , C. sakazakii and L. monocytogenes and to characterise the potential risk of these pathogens in raw milk and cheese. The raw milk ( n  = 25) and cheese samples ( n  = 20) were analysed by multiplex RT i‐ PCR assay and detected for quantification of the three pathogens. In this study, B. melitensis , C. sakazakii and L. monocytogenes were simultaneously identified using BMEII 0466 , mms operon and hly as target genes, respectively. The multiplex RT i‐ PCR assay that was developed showed good sensitivity and selectivity for the pathogenic microorganisms ( r 2  = 0.986–0.997). Multiplex RT i‐ PCR results showed that most of the samples were contaminated with the pathogens screened.

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