Use of advanced molecular biology techniques for detailed characterization of probiotic bacteria from a dietary supplement

Cílem diplomové práce byl průkaz probiotických bakterií z probiotického doplňku stravy ve formě sirupu. Z komplexní matrice výrobku byla izolována DNA v dostatečné čistotě a kvalitě vhodné pro metodu PCR v reálném čase a následně PCR-HRM. K amplifikaci pomocí PCR v reálném čase a PCR-HRM byly použity specifické primery pro rody Lactobacillus, Bifidobacterium a Streptococcus a na úrovni druhu pro druhy Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus reuteri, Bifidobacterium animalis, Bifidobacterium breve, Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium infantis a Streptococcus thermophilus. Po optimalizaci podmínek pro specifické primery byly ve výrobku prokázány všechny deklarované bakterie.The aim of the diploma thesis was the identification of probiotic bacteria from a probiotic food supplement in the form of syrup. DNA was isolated from the complex matrix of the product in sufficient purity and quality suitable for the real-time PCR method followed by PCR-HRM. Specific primers for the genera Lactobacillus, Bifidobacterium and Streptococcus and at the species level for the species Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus reuteri, Bifidobacterium animalis, Bifidobacterium breve, Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium infantis and Streptococcus thermophilus were used for amplification by real-time PCR and PCR-HRM. After optimizing the conditions for specific primers, all declared bacteria were detected in the product.