TY - JOUR
T1 - IS-pro
T2 - High-throughput molecular fingerprinting of the intestinal microbiota
AU - Budding, A. E.
AU - Grasman, M. E.
AU - Lin, F.
AU - Bogaards, J. A.
AU - Soeltan-Kaersenhout, D. J.
AU - Vandenbroucke-Grauls, C. M. J. E.
AU - van Bodegraven, A. A.
AU - Savelkoul, P. H. M.
PY - 2010/11
Y1 - 2010/11
N2 - 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. © FASEB.
AB - 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. © FASEB.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=78649883215&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/20643909
U2 - https://doi.org/10.1096/fj.10-156190
DO - https://doi.org/10.1096/fj.10-156190
M3 - Article
C2 - 20643909
SN - 0892-6638
VL - 24
SP - 4556
EP - 4564
JO - FASEB Journal
JF - FASEB Journal
IS - 11
ER -