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Analysis of the composition of selected probiotic products by PCR-HRM
Tomanová, Barbora ; Španová, Alena (referee) ; Trachtová, Štěpánka (advisor)
This work was focused on the detection of probiotic bacteria in four different probiotic products (probiotic cream, probiotic tampons, oral probiotics and soy beverages with probiotics). The viability of the bacteria contained in the products was verified. Complex matrices of the products were used to isolate DNA in a quality suitable for the PCR method, followed by identification of the declared bacterial genus and species. Amplification was achieved with conventional PCR and real-time PCR, genus- and species-specific primers were used. Bacteria, of the genus Lactobacillus and Bacillus and bacterial species Lactobacillus pentosus, Lactobacillus rhamnosus, Lactobacillus fermentum and Lactobacillus gasseri, were proven to be within the products. Subsequently, the DNA from mixed bacterial species in the probiotic tampon were distinguished using PCR-HRM. Five sets of primers were used to test this. Two sets of primers (primers P1V1, P2V1 and V1F-HRM, V1R-HRM) were evaluated as the most suitable for resolution.
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Real-time PCR and it´s use in food processing
Tomanová, Barbora ; Pravečková, Martina (referee) ; Trachtová, Štěpánka (advisor)
Polymerase chain reaction (PCR) is a method abundantly used in molecular diagnostics. PCR in real time or quantitative PCR (qPCR) is one of its modifications and thanks to its advantages it finds still wider utilization nowadays. It finds its use in the food-processing industry too with relatively precise detection, identification, and qualification of both desirable and undesirable components in food, which often brings considerable difficulties and leads to an intensive development of this method. In the experimental part a DNA was isolated from dairy product Bio Via Natur drink for its further processing by means of PCR and gain more detailed information about a bacterial composition using real-time PCR and HRM analysis.
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Analysis of the composition of selected probiotic products by PCR-HRM
Tomanová, Barbora ; Španová, Alena (referee) ; Trachtová, Štěpánka (advisor)
This work was focused on the detection of probiotic bacteria in four different probiotic products (probiotic cream, probiotic tampons, oral probiotics and soy beverages with probiotics). The viability of the bacteria contained in the products was verified. Complex matrices of the products were used to isolate DNA in a quality suitable for the PCR method, followed by identification of the declared bacterial genus and species. Amplification was achieved with conventional PCR and real-time PCR, genus- and species-specific primers were used. Bacteria, of the genus Lactobacillus and Bacillus and bacterial species Lactobacillus pentosus, Lactobacillus rhamnosus, Lactobacillus fermentum and Lactobacillus gasseri, were proven to be within the products. Subsequently, the DNA from mixed bacterial species in the probiotic tampon were distinguished using PCR-HRM. Five sets of primers were used to test this. Two sets of primers (primers P1V1, P2V1 and V1F-HRM, V1R-HRM) were evaluated as the most suitable for resolution.
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Real-time PCR and it´s use in food processing
Tomanová, Barbora ; Pravečková, Martina (referee) ; Trachtová, Štěpánka (advisor)
Polymerase chain reaction (PCR) is a method abundantly used in molecular diagnostics. PCR in real time or quantitative PCR (qPCR) is one of its modifications and thanks to its advantages it finds still wider utilization nowadays. It finds its use in the food-processing industry too with relatively precise detection, identification, and qualification of both desirable and undesirable components in food, which often brings considerable difficulties and leads to an intensive development of this method. In the experimental part a DNA was isolated from dairy product Bio Via Natur drink for its further processing by means of PCR and gain more detailed information about a bacterial composition using real-time PCR and HRM analysis.
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