title: Use of 16S-23S rRNA spacer-region (SR)-PCR for identification of intestinal clostridia.
abstract: The suitability of a species identification technique based on PCR analysis of 16S-23S rRNA spacer region (SR) polymorphism for human intestinal Clostridium species was evaluated. This SR-PCR based technique is highly reproducible and successfully differentiated the strains tested, which included 17 ATCC type strains of Clostridium and 152 human stool Clostridium isolates, at the species or intraspecies level. Ninety-eight of 152 stool isolates, including C. bifermentans, C. butyricum, C. cadaveris, C. orbiscindens, C. paraputrificum, C. pefringens, C. ramosum, C. scindens, C. spiroforme, C. symbiosum and C. tertium, were identified to species level by SR-PCR patterns that were identical to those of their corresponding ATCC type strains. The other 54 stool isolates distributed among ten SR-PCR patterns that are unique and possibly represent ten novel Clostridium species or subspecies. The species identification obtained by SR-PCR pattern analysis completely agreed with that obtained by 16S rRNA sequencing, and led to identification that clearly differed from that obtained by cellular fatty acid analysis for 23/152 strains (15%). These results indicate that SR-PCR provides an accurate and rapid molecular method for the identification of human intestinal Clostridium species.
authors.affiliation: Research Service, VA Medical Center West Los Angeles, CA 90073, USA. yulis1@yahoo.com
authors.forename: Yuli
authors.initials: Y
authors.lastname: Song
authors.forename: Chengxu
authors.initials: C
authors.lastname: Liu
authors.forename: Denise
authors.initials: D
authors.lastname: Molitoris
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Tomzynski
authors.forename: Maureen
authors.initials: M
authors.lastname: Mc Teague
authors.forename: Erik
authors.initials: E
authors.lastname: Read
authors.forename: Sydney M
authors.initials: SM
authors.lastname: Finegold
mesh_terms: D001483:Base Sequence; D003013:Clostridium / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D003015:Clostridium Infections / Q000382:microbiology*; D021901:DNA, Intergenic / Q000235:genetics; D005227:Fatty Acids / Q000032:analysis; D005243:Feces / Q000382:microbiology; D006801:Humans; D007410:Intestinal Diseases / Q000382:microbiology*; D007422:Intestines / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics
chemical_list: D021901:DNA, Intergenic; D005227:Fatty Acids; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S