Evaluation of using a short region of the recA gene for rapid and sensitive speciation of dominant bifidobacteria in the human large intestine |
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| Institution: | 1. Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran;2. Department of Medicine, University of Alberta, Edmonton, AB, Canada;3. Legal Medicine Research Center, Legal Medicine Organization, Tabriz, Iran;4. School of Health and Nutrition, Tabriz University of Medical Sciences, Tabriz, Iran;1. Neurodegeneration Imaging Group, Department of Basic and Clinical Neuroscience, Institute of Psychiatry, Psychology and Neuroscience (IoPPN), King''s College London, London, UK;2. Division of Brain Sciences, Imperial College London, Hammersmith Hospital, London, UK;3. Department of Neuroimaging, Institute of Psychiatry, King''s College London, London, UK;4. Parkinson''s Centre of Excellence, Department of Neurology, King''s College Hospital Foundation Trust, UK |
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| Abstract: | The feasibility of intragenerically characterizing bifidobacteria by a comparison of a short region within the recA gene was tested. An ∼300 bp fragment of the recA gene was PCR-amplified from six species from the genus Bifidobacterium using primers directed to two universally conserved regions of the recA gene. A phylogenetic analysis of the sequenced recA products compared favorably to classification based on the 16S rRNA sequences of the species tested. To apply this rapid methodology to unknown human intestinal bifidobacteria, 46 isolates were randomly chosen from the feces of four subjects and initially characterized by RFLP analysis of a PCR-amplified region of their 16S RNA genes. From a representative of the dominant RFLP family in each of the subjects, the recA segment was PCR-amplified, sequenced and phylogenetically analyzed. All four isolates were found to be related to one another and to B. longum and B. infantis. These results illustrate that the recA gene may be useful for intrageneric phylogenetic analysis as well as for the identification of unknown fecal bifidobacteria. |
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