A new rapid real-time PCR method for detection of listeria monocytogenes targeting the hlyA gene

Pei Liu, Hiromi Mizue, Kumiko Fujihara, Hiroshi Kobayashi, Hideaki Kamikado, Takashi Tanaka, Ken-ichi Honjoh, Takahisa Miyamoto

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

In this study, primer sets of L. monocytogenes virulence gene hlyA were designed for the L. monocytogenes- specific PCR assay. To evaluate the performance of these primer sets, the detection sensitivity and specificity were examined and the most suitable primer set was selected. Subsequently, it was subjected to compare the performance for detection of L. monocytogenes with commercially available kits (TaKaRa detection kit and ABI detection kit) on DNA level. Results of real-time PCR showed that the efficiency of this new primer set was 101.6% while TaKaRa and ABI detection kit were 101.1% and 107.4%, respectively. The detection sensitivity of all three methods was equivalent to less than 1 copy per reaction using purified genomic DNA as template. Detection specificity were 100% when testing L. monocytogenes isolates, and for other Listeria isolates were 15.4%, 76.9% and 100% by the method using hlyA L. monocytogenes detection primer set 7, TaKaRa and ABI detection kit, respectively. In summary, this new PCR detection method is relatively sensitive and more specific to L. monocytogenes under certain conditions, could serve as a rapid detection method and has the potential for detection of L. monocytogenes from contaminated food.

Original languageEnglish
Pages (from-to)47-57
Number of pages11
JournalFood Science and Technology Research
Volume18
Issue number1
DOIs
Publication statusPublished - Jan 1 2012

Fingerprint

Listeria
Gene Targeting
Listeria monocytogenes
gene targeting
Real-Time Polymerase Chain Reaction
quantitative polymerase chain reaction
DNA
Genes
Assays
Testing
methodology
Polymerase Chain Reaction
Gene
Targeting
Virulence
Sensitivity and Specificity
Food
food contamination

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Chemical Engineering(all)
  • Industrial and Manufacturing Engineering
  • Marketing

Cite this

A new rapid real-time PCR method for detection of listeria monocytogenes targeting the hlyA gene. / Liu, Pei; Mizue, Hiromi; Fujihara, Kumiko; Kobayashi, Hiroshi; Kamikado, Hideaki; Tanaka, Takashi; Honjoh, Ken-ichi; Miyamoto, Takahisa.

In: Food Science and Technology Research, Vol. 18, No. 1, 01.01.2012, p. 47-57.

Research output: Contribution to journalArticle

Liu, Pei ; Mizue, Hiromi ; Fujihara, Kumiko ; Kobayashi, Hiroshi ; Kamikado, Hideaki ; Tanaka, Takashi ; Honjoh, Ken-ichi ; Miyamoto, Takahisa. / A new rapid real-time PCR method for detection of listeria monocytogenes targeting the hlyA gene. In: Food Science and Technology Research. 2012 ; Vol. 18, No. 1. pp. 47-57.
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