Detection and Identification of Probiotic Lactobacillus plantarum Strains by Multiplex PCR Using RAPD-Derived Primers.

Alex Galanis,Nikos Chorianopoulos,Yiannis Kourkoutas,Chrysoula C Tassou

doi:10.3390/ijms161025141

Abstract

Lactobacillus plantarum 2035 and Lactobacillus plantarum ACA-DC 2640 are two lactic acid bacteria (LAB) strains that have been isolated from Feta cheese. Both display significant potential for the production of novel probiotic food products. The aim of the present study was the development of an accurate and efficient method for the molecular detection and identification of the above strains in a single reaction. A multiplex PCR assay was designed for each strain, based on specific primers derived from Random Amplified Polymorphic DNA (RAPD) Sequenced Characterized Amplified Region (SCAR) analysis. The specificity of the assay was tested with a total of 23 different LAB strains, for L. plantarum 2035 and L. plantarum ACA-DC 2640. The multiplex PCR assay was also successfully applied for the detection of the above cultures in yogurt samples prepared in our lab. The proposed methodology may be applied for monitoring the presence of these strains in food products, thus evaluating their probiotic character. Moreover, our strategy may be adapted for other novel LAB strains with probiotic potential, thus providing a powerful tool for molecular discrimination that could be invaluable to the food industry.

Highlights

Probiotics are living microorganisms, with bifidobacteria and lactic acid bacteria (LAB)constituting the most common microbial groups
The theory that regular consumption of lactic acid bacteria in fermented foods may contribute to enhancing health and longevity was originally developed by the Russian immunologist and Nobel Laureate in Medicine Elie Metchnikoff and was presented in his book “The prolongation of life”, published in 1907
A probiotic-rich diet has been connected with the prevention and potential treatment of several digestive disorders, such as irritable bowel syndrome [5], necrotizing enterocolitis (NEC) in neonates [6], and ulcerative colitis [7]

Summary

Introduction

Probiotics are living microorganisms, with bifidobacteria and lactic acid bacteria (LAB)constituting the most common microbial groups. 6.8% from 2013 to 2018 [3] Their strong market development and immense future potential are mainly due to numerous studies that have linked consumption of probiotic beverages, foods, and supplements to beneficial effects for consumers’ health [4]. Recent studies employing in vitro and in vivo systems have indicated that probiotic treatment might reduce the risk for colon, liver, and bladder cancers [8]. It may be effective against diet-induced obesity through the modulation of genes associated with metabolism and inflammation in the liver and adipose tissue [9], and even be therapeutically beneficial for the treatment of anxiety and depression [10]

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