Identification of bacterial species in complex probiotic products using multiplex PCR and flow cytometry
Abstract
Probiotics are a group of microorganisms known for their health promoting effects. Consumer demand for probiotic products containing multiple species has increased due to the belief that their synergistic effects exert more health benefits. This puts additional pressure on a quality control laboratory who are required to confirm the identification of each organism in a product, rendering molecular methods requiring the cultivation of bacteria unsuitable due to long lead times of results. Therefore, identification using a culture-independent technique is required. Two methods were investigated: multiplex PCR and flow cytometry. Multiplex PCR can detect individual species within a mixed population using species-specific primer pairs. The aim of this study was to develop a multiplex PCR assay capable of identifying Lactobacillus acidophilus, Lacticaseibacillus paracasei, Lacticaseibacillus rhamnosus, Bifidobacterium animalis subsp. lactis and Bifidobacterium bifidum in mixed populations. Additional work was also carried out to determine if L. acidophilus could be detected in a mixed product with the use of polyclonal antibodies using flow cytometry, which provided promising preliminary data. This study culminated in the production and optimisation of a multiplex PCR assay capable of detecting five species within a mixed product, reducing result lead times and expenseSimilar works
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Last time updated on 20/05/2024
This paper was published in Cronfa at Swansea University.
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