期刊名称：Annals of the University Dunarea de Jos of Galati. Fascicle VI : Food Technology
印刷版ISSN：1843-5157
电子版ISSN：2068-259X
出版年度：2019
卷号：43
期号：2
页码：1-12
出版社：Galati University Press
摘要：In this study conventional PCR and multiplex PCR based method was
developed for the detection of two common foodborne pathogens Salmonella
enterica and Listeria monocytogenes in 20 raw milk samples. The PCR was
undertaken to detect two genes namely, Salmonella enterotoxin (stn) gene and
phosphatidylinositol-specific phospholipase C gene (plcA) from both the
organisms. The DNA templates (for both organisms) were amplified using
specific set of primers. The resulting amplicons were found to be 265 bp and
147 bp respectively. Validation studies were further performed in artificial
spiked milk samples and raw milk samples using multiplex PCR. The available
detection methods are bacterial culturing, biochemical tests, serological tests,
antibiotic sensitivity, ELISA and PCR. All these methods are either expensive
or non-confirmatory and have some limitations. The reported multiplexed PCR
based genetic marker completes overall analysis in 80 min which is the
minimum time reported so far for the confirmation of these foodborne
pathogens. Sensitivity and specificity of developed method was calculated and
compared with different conventional methods. The detection limit of the assay
for the S. enterica was 6.6 x100 CFU/mL and for L. monocytogenes was
4.5x100 CFU/mL.
关键词：Salmonella enterica; Listeria monocytogenes; raw milk; conventional
PCR; multiplex PCR
