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A PCR-based method for identification of bifidobacteria from the human alimentary tract at the species level

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Author: Venema, K. · Maathuis, A.J.H.
Type:article
Date:2003
Institution: TNO Voeding
Source:FEMS Microbiology Letters, 1, 224, 143-149
Identifier: 237187
doi: doi:10.1016/S0378-1097(03)0043611
Keywords: Health Nutrition · Physiological Sciences · 16S rDNA · Alimentary tract · Amplified ribosomal DNA restriction analysis · Bifidobacterium · bacterial DNA · bacterial enzyme · bacterial protein · DNA 16S · restriction endonuclease · ribosome protein · amplified ribosomal DNA restriction analysis · article · bacterial strain · bacterium identification · bacterium isolate · Bifidobacterium · controlled study · digestive system · DNA fingerprinting · feces analysis · human · human tissue · in vitro study · large intestine · nonhuman · nucleotide sequence · polymerase chain reaction · priority journal · restriction fragment · Bifidobacterium · Child · DNA, Bacterial · Feces · Humans · Intestine, Large · Polymerase Chain Reaction · Restriction Mapping · RNA, Ribosomal, 16S · Bifidobacterium

Abstract

A polymerase chain reaction (PCR)-based method was developed for the identification of isolates of Bifidobacterium at the species level. Using two Bifidobacterium-specific primers directed against the 16S ribosomal gene (Bif164 and Bif662), a PCR product was obtained from the type strains of 12 different bifidobacterial species that have been found in the human alimentary tract. After purification of the PCR products, the DNA was restricted with five different restriction enzymes. The size of the different restriction fragments was used as a fingerprint for the identification of individual bifidobacterial species. The amplified ribosomal DNA restriction analysis method was subsequently used to speciate bifidobacterial isolates from child's feces and from an in vitro model of the large intestine. © 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.