Detection and Identification of Probiotic Lactobacillus plantarum Strains by Multiplex PCR Using RAPD-Derived Primers.
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.
Department of Molecular Biology and Genetics, Democritus University of Thrace, Alexandroupolis 68100, Greece. agalanis@mbg.duth.gr.
Alex
A
Galanis
Department of Molecular Biology and Genetics, Democritus University of Thrace, Alexandroupolis 68100, Greece. ikourkou@mbg.duth.gr.
Yiannis
Y
Kourkoutas
Institute of Technology of Agricultural Products, Hellenic Agricultural Organization-DEMETER, Sof. Venizelou 1, Lycovrissi, Attiki 14123, Greece. ctassou@nagref.gr.
Chrysoula C
CC
Tassou
Institute of Technology of Agricultural Products, Hellenic Agricultural Organization-DEMETER, Sof. Venizelou 1, Lycovrissi, Attiki 14123, Greece. nchorian@nagref.gr.
Nikos
N
Chorianopoulos
D001483:Base Sequence; D002611:Cheese / Q000382:microbiology; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D048191:Lactiplantibacillus plantarum / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D058889:Molecular Typing / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D019936:Probiotics / Q000737:chemistry*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*; D017422:Sequence Analysis, DNA; D015014:Yogurt / Q000382:microbiology
D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S
Lactobacillus plantarum; RAPD; SCAR; lactic acid bacteria; multiplex PCR; probiotics