ISSN: 0973-7510

E-ISSN: 2581-690X

Research Article | Open Access
Rajashree Jena1,2, Prasanta Kumar Choudhury1,2 , Anil Kumar Puniya1 and Sudhir Kumar Tomar1
1Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal – 132 001, Haryana, India.
2Department of Dairy Technology, School of Agricultural and Bioengineering, Centurion University of Technology and Management, Paralakhemundi – 761 211, Odisha, India.
J Pure Appl Microbiol. 2022;16(1):503-513 | Article Number: 6994
https://doi.org/10.22207/JPAM.16.1.48 | © The Author(s). 2022
Received: 19/04/2021 | Accepted: 27/01/2022 | Published online: 21/02/2022
Issue online: March 2022
Abstract

Bifidobacteria are widely used as probiotics for their application in the development of functional food and prophylactic therapy. This has necessitated the development of a molecular approach for the genera to be widely identified up to species and subspecies level. In the current study, PCR-RFLP of the partial RNA polymerase β-subunit (rpoB) gene fragment was evaluated for differential identification of Bifidobacterium species. The rpoB gene partial sequences of 575 bp were amplified from 93 previously identified isolates collected from various sources of human and animal origin along with 12 standard reference strains. The PCR amplified products were digested with three restriction endonucleases HhaI, HinfI and BanI separately. Dendrograms constructed from the patterns of HhaI, were found to be more discriminatory and successfully differentiated all the twelve species and also at sub-species level in between B. longum subsp. longum and B. longum subsp. infantis. However, B. adolescentis and B. pseudocatenulatum group clusters were not separated and represented by one group. The groups were further discriminated by HinfI restriction digestion. A separate combination thereof may be used for inferring the classification of bifidobacterial species targeted on rpoB PCR-RFLP analysis. To our knowledge, this work is the first report based on use of rpoB PCR-RFLP for discrimination of the isolates of genus Bifidobacterium and also provides insights into specific advantages of this method over hsp60 PCR-RFLP in differentiating B. longum subsp. longum and B. longum subsp. infantis.

Keywords

Bifidobacterium, PCR-RFLP, HhaI, rpoB gene, probiotics

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