The development of a DNA microarray-based assay for the characterization of commercially formulated microbial products.
Commercially formulated bioproducts containing a complex consortia of bacteria as an active ingredient pose a significant challenge for regulatory agencies and companies seeking to assess the safety and efficacy of these bioproducts. The main challenge stems from how to characterize the bacterial composition of these products, for which there is presently a lack of suitable methods. A prototype DNA microarray composed of oligonucleotide probes for functional genes, virulence factors, and taxonomic genes for a number of bacterial species was developed to examine the utility of microarray technology as a molecular tool for characterizing consortia bioproducts. The genomic DNA from four different products was extracted by two methods and examined with the microarray prototype and by denaturing gradient gel electrophoresis (DGGE). Although the identity of the consortial species remains unknown, the microarray assay provided unique and reproducible hybridization patterns for all four products, and agreed with the fingerprints generated by DGGE. The ability to differentiate between a variety of consortia products demonstrates that DNA microarrays have the potential to be a powerful tool in monitoring complex microbial communities.
Dubois
J W
JW
Environmental Sector, Biotechnology Research Institute, NRC, 6100 Royalmount Avenue, Montreal, Quebec, Canada H4P 2R2.
Hill
S
S
England
L S
LS
Edge
T
T
Masson
L
L
Trevors
J T
JT
Brousseau
R
R
D001431:Bacteriological Techniques; D001483:Base Sequence; D001688:Biological Products / Q000032:analysis* / Q000235:genetics*; D016172:DNA Fingerprinting; D015342:DNA Probes / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction
D001688:Biological Products; D015342:DNA Probes; D004269:DNA, Bacterial