High taxonomic level fingerprint of the human intestinal microbiota by ligase detection reaction--universal array approach.
BACKGROUND
Affecting the core functional microbiome, peculiar high level taxonomic unbalances of the human intestinal microbiota have been recently associated with specific diseases, such as obesity, inflammatory bowel diseases, and intestinal inflammation.


RESULTS
In order to specifically monitor microbiota unbalances that impact human physiology, here we develop and validate an original DNA-microarray (HTF-Microbi.Array) for the high taxonomic level fingerprint of the human intestinal microbiota. Based on the Ligase Detection Reaction-Universal Array (LDR-UA) approach, the HTF-Microbi.Array enables specific detection and approximate relative quantification of 16S rRNAs from 30 phylogenetically related groups of the human intestinal microbiota. The HTF-Microbi.Array was used in a pilot study of the faecal microbiota of eight young adults. Cluster analysis revealed the good reproducibility of the high level taxonomic microbiota fingerprint obtained for each of the subject.


CONCLUSIONS
The HTF-Microbi.Array is a fast and sensitive tool for the high taxonomic level fingerprint of the human intestinal microbiota in terms of presence/absence of the principal groups. Moreover, analysis of the relative fluorescence intensity for each probe pair of our LDR-UA platform can provide estimation of the relative abundance of the microbial target groups within each samples. Focusing the phylogenetic resolution at division, order and cluster levels, the HTF-Microbi.Array is blind with respect to the inter-individual variability at the species level.
Candela
Marco
M
Department of Pharmaceutical Science, University of Bologna, Italy. marco.candela@unibo.it
Consolandi
Clarissa
C
Severgnini
Marco
M
Biagi
Elena
E
Castiglioni
Bianca
B
Vitali
Beatrice
B
De Bellis
Gianluca
G
Brigidi
Patrizia
P
D000328:Adult; D016000:Cluster Analysis; D016172:DNA Fingerprinting / Q000379:methods*; D005243:Feces / Q000382:microbiology; D006801:Humans; D007422:Intestines / Q000382:microbiology*; D054892:Metagenome; D056186:Metagenomics / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D015203:Reproducibility of Results
D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S
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