Identification and phylogenetic analysis of Lactobacillus using multiplex RAPD-PCR

Multiplex RAPD-PCR was used to generate unique and identifying DNA profiles for isolates of the genus Lactobacillus. The method that was used was based on the combination of two 10-mer oligonucleotides in a single PCR. The generated RAPD profiles enabled discrimination of all lactobacillus strains that were used in this study. A dendrogram was generated from the RAPD profiles. The results of genetic relatedness obtained from the dendrogram were compared with the results obtained using carbohydrate fermentation profiles. Most of the gastrointestinal isolates studied could not be grouped using carbohydrate fermentation profiles. The RAPD profiles provided sufficient information to prepare a dendrogram of genetic relatedness. The gastrointestinal isolates were clustered together on the dendrogram. Furthermore an isolate originating from the stomach (strain ML004) was closely related to Lactobacillus fermentum. It was concluded that multiplex RAPD-PCR was useful for characterisation and inference of relatedness of Lactobacillus isolates.