title: IS-pro: high-throughput molecular fingerprinting of the intestinal microbiota.
abstract: The human intestinal microbiota is known to play an important role in human health and disease, and with the advent of novel molecular techniques, disease-specific variations in its composition have been found. However, analysis of the intestinal microbiota has not yet been applicable in large-scale clinical research or routine diagnostics because of the complex and expensive nature of the techniques needed. Here, we describe a new PCR-based profiling technique for high-throughput analysis of the human intestinal microbiota, which we have termed IS-pro. This technique combines bacterial species differentiation by the length of the 16S-23S rDNA interspace region with instant taxonomic classification by phylum-specific fluorescent labeling of PCR primers. We validated IS-pro in silico, in vitro, and in vivo, on human colonic biopsies and feces, and introduced a standardized protocol for data analysis. IS-pro is easy to implement in general clinical microbiological laboratories with access to capillary gel electrophoresis, and the high-throughput nature of the test makes analysis of large numbers of samples feasible. This combination renders IS-pro ideally suited for use in clinical research and routine diagnostics.
authors.affiliation: Department of Medical Microbiology and Infection Control, VU University Medical Center, Amsterdam, The Netherlands. d.budding@vumc.nl
authors.forename: A E
authors.initials: AE
authors.lastname: Budding
authors.forename: M E
authors.initials: ME
authors.lastname: Grasman
authors.forename: F
authors.initials: F
authors.lastname: Lin
authors.forename: J A
authors.initials: JA
authors.lastname: Bogaards
authors.forename: D J
authors.initials: DJ
authors.lastname: Soeltan-Kaersenhout
authors.forename: C M J E
authors.initials: CM
authors.lastname: Vandenbroucke-Grauls
authors.forename: A A
authors.initials: AA
authors.lastname: van Bodegraven
authors.forename: P H M
authors.initials: PH
authors.lastname: Savelkoul
mesh_terms: D001419:Bacteria / Q000145:classification / Q000235:genetics*; D044822:Biodiversity; D016172:DNA Fingerprinting / Q000379:methods*; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D019075:Electrophoresis, Capillary / Q000379:methods; D005243:Feces / Q000382:microbiology*; D006801:Humans; D007422:Intestines / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D013045:Species Specificity
chemical_list: D021903:DNA, Ribosomal Spacer