Rapid Detection and Identification of Probiotic <i>Lactobacillus casei</i> ATCC 393 by Multiplex PCR

Abstract

Many functional foods containing probiotic strains have been developed recently. Lactobacillus casei ATCC 393 is one of the most frequently used cultures in probiotic products. The present study aimed to develop a method for the detection and identification of L. casei ATCC 393 based on genetic polymorphisms of the hsp60 gene. A multiplex polymerase chain reaction (PCR) assay was designed, utilizing two novel strain-specific primer sets that enable identification of L. casei ATCC 393.The accuracy of our method was further confirmed by successful identification of our strain in probiotic cheese. The method described is an easy to use, rapid, inexpensive and accurate tool that may be readily applied to food, fecal and intestinal samples.

Effect of immobilized Lactobacillus casei on the evolution of flavor compounds in probiotic dry-fermented sausages during ripening

“…The presence of the probiotic strain at levels above the minimum concentration for conferring a probiotic effect (≥6 log cfu/g) at the end of the ripening process and after mild heat treatment was confirmed by an efficient molecular tool (Karapetsas, Vavoulidis, Galanis, Sandaltzopoulos, & Kourkoutas, 2010; Meat Science 100 (2015) 41-51 Sidira et al, 2014). Noticeably, the probiotic properties of free and immobilized L. casei ATCC 393 were previously assessed by documenting maintenance of cell viability after transit through the GI tract, adhesion at the large intestine and regulation of the intestinal microbial flora in rats (Saxami et al, 2012;Sidira et al, 2010).…”

Effect of Probiotic-Fermented Milk Administration onGastrointestinal Survival of <i>Lactobacillus casei</i> ATCC 393 and Modulation of Intestinal Microbial Flora

“…1 ). Identification of L. casei ATCC 393 after growth on MRS agar plates was carried out by multiplex PCR assay using specific primers that generated PCR products of 67 and 144 bp [Karapetsas et al, 2010]. A set of universal for lactobacilli primers generating a PCR product of 340 bp was used as positive control [Walter et al, 2001].…”

“…Molecular identification of L. casei ATCC 393 strain in fecal samples was carried out as described recently [Karapetsas et al, 2010]. Briefly, genomic DNA from lactobacilli suspensions prepared after microbiological analysis as described above, was extracted using a DNeasy Tissue Kit (Qiagen, Hilden, Germany) according to the manufacturer's protocol.…”

“…Multiplex PCR reactions were carried out in a total volume of 50 l, containing 5 units Taq DNA polymerase (HyTest Ltd), 400 M of each dNTP (Promega), 1.5 m M MgCl 2 (HyTest Ltd) and 100 ng template DNA. The specific for L. casei ATCC 393 primers that recognize the hsp60 gene consisted of GGCGACCAAGGCAGCG (10 pmol), CTTCG-GTTTCATCTTCC (50 pmol) and GGCCAACTTTTTCCATA (50 pmol) [Karapetsas et al, 2010]. Additionally, a set of primers that recognize the 16S rRNA gene of all Lactobacillus strains was used as positive control.…”

“…Among LAB, Lactobacillus casei ATCC 393 strain has been extensively incorporated into food products [Kourkoutas et al, 2005[Kourkoutas et al, , 2006aLi et al, 2009]. Recently, a new method for rapid detection and identification of the above strain based on multiplex PCR was demonstrated [Karapetsas et al, 2010]. The presented methodology proved an efficient tool for accurate, convenient and reliable identification of the L. casei ATCC 393 in food products.…”