Novel PCR Assays Complement Laser Biosensor-Based Method and Facilitate Listeria Species Detection from Food

Kwang-Pyo Kim; Atul K Singh; Xingjian Bai; Lena Leprun; Arun K Bhunia

doi:10.3390/s150922672

Novel PCR Assays Complement Laser Biosensor-Based Method and Facilitate Listeria Species Detection from Food

Sensors (Basel). 2015 Sep 8;15(9):22672-91. doi: 10.3390/s150922672.

Authors

Kwang-Pyo Kim^{1

2}, Atul K Singh³, Xingjian Bai⁴, Lena Leprun⁵, Arun K Bhunia^{6

7}

Affiliations

¹ Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN 47907, USA. kpkim@jbnu.ac.kr.
² Department of Food Science and Technology, College of Agriculture and Life Sciences, Chonbuk National University, Jeonbuk 561756, Korea. kpkim@jbnu.ac.kr.
³ Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN 47907, USA. aksingh@purdue.edu.
⁴ Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN 47907, USA. bai16@purdue.edu.
⁵ Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN 47907, USA. l.leprun@laposte.net.
⁶ Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, West Lafayette, IN 47907, USA. bhunia@purdue.edu.
⁷ Department of Comparative Pathobiology, Purdue University, West Lafayette, IN 47907, USA. bhunia@purdue.edu.

Abstract

The goal of this study was to develop the Listeria species-specific PCR assays based on a house-keeping gene (lmo1634) encoding alcohol acetaldehyde dehydrogenase (Aad), previously designated as Listeria adhesion protein (LAP), and compare results with a label-free light scattering sensor, BARDOT (bacterial rapid detection using optical scattering technology). PCR primer sets targeting the lap genes from the species of Listeria sensu stricto were designed and tested with 47 Listeria and 8 non-Listeria strains. The resulting PCR primer sets detected either all species of Listeria sensu stricto or individual L. innocua, L. ivanovii and L. seeligeri, L. welshimeri, and L. marthii without producing any amplified products from other bacteria tested. The PCR assays with Listeria sensu stricto-specific primers also successfully detected all species of Listeria sensu stricto and/or Listeria innocua from mixed culture-inoculated food samples, and each bacterium in food was verified by using the light scattering sensor that generated unique scatter signature for each species of Listeria tested. The PCR assays based on the house-keeping gene aad (lap) can be used for detection of either all species of Listeria sensu stricto or certain individual Listeria species in a mixture from food with a detection limit of about 10⁴ CFU/mL.

Keywords: BARDOT; Listeria adhesion protein; Listeria species; PCR; alcohol acetaldehyde dehydrogenase; detection.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Biosensing Techniques / methods*
DNA, Bacterial / analysis
DNA, Bacterial / genetics
Food Microbiology / methods*
Limit of Detection
Listeria / genetics*
Listeria / isolation & purification*
Molecular Typing / methods*
Polymerase Chain Reaction / methods*
Principal Component Analysis

Substances

DNA, Bacterial