You can send isolated strains on plates or slants. We do the following identifications:
GC-FAME (Gas Chromatographic Fatty Acid Methyl Ester)
Also known as Cellular Fatty Acid Analysis, allows for bacterial or
yeast identification based on the specific fatty acid composition of
the cell wall. Fatty acids are extracted from cultured samples and are
separated by gas chromatography. A computer generated, unique profile
pattern of the extracted fatty acids (9 to 20 carbon chains long) is
compared through pattern recognition programs, to our microbial
databases of well over 2600 species and subspecies. Our reports
include the fatty acid profiles, a listing of the best database
matches, along with an assigned statistical probability value
indicating the confidence level of the match.
Biolog® MicroLog
This automated, metabolic characterization system uses a 96-well
microtiter plate arrayed with different carbon sources and the
microorganism of interest. Each microorganism has unique capacities to
oxidize the various carbon sources. Tetrazolium dye in each of the
wells turn darker shades of purple as the carbon sources are oxidized
by the microorganism. The test yields a characteristic pattern of
positive (purple) and negative wells which provide a metabolic
signature of the inoculated organism. The system's computer the
pattern signature with the database of interest to determine an
identification. Databases include aerobes, anaerobes, gram positive
& negative bacteria, yeast, actinomycetes, and lactic acid
bacteria.
RiboPrinter™ (Qualicon™, a DuPont subsidiary)
The RiboPrinter™ system characterized bacteria by creating a
fingerprint pattern of an isolate's genetic material. Pure colonies
are isolated from culture, inactivated by heat and loaded into the
sample carrier. DNA is extracted from the cell lysate and is digested
to completion using the EcoR1 restriction enzyme. The DNA fragments
are separated according to molecular size by gel electrophoresis and
are the transferred to moving Nylon membranes. After denaturation,
each membrane is hybridized with a chemically labeled rRNA operon probe
from E. coli. The membrane is washed and treated with a blocking
buffer and an antisulfonated, DNA antibody/alkaline phosphatase
conjugate. Unbound conjugate is removed and a chemiluminescent
substrate applied. This step makes each electrophoresis band
containing genetic information from the rRNA genes visible to a custom
CCD camera in the RiboPrinter™ system. The camera detects the light
intensity of the targeted bands and converts the patterns from
luminescent DNA fragments to digital information. Using algorithms, a
pattern is extracted from the image data and is compared to other known
patterns stored in the database to characterize and identify each
sample.
MicroSeq™ 16S rRNA Gene Sequencing (PE Applied Biosystems)
Genomic DNA is extracted directly from dead or alive bacterial
colonies grown under any conditions. The 16S rRNA gene is amplified
using universal primers and thermalcyclers. The amplified 16S rRNA
gene product is sequenced using dye terminator cycle sequencing
chemistry. The sequence reactions are analyzed using automated DNA
sequencers and software. Unknown bacteria samples are identified using
the MicroSeq™ microbial identification software and 16S rRNA gene
sequence database, containing over 1,500 entries. Dependant upon the
level of confidence, speed and cost desired, either the entire 1540
base pair 16S rRNA gene, or a smaller portion of the gene can be (500
bp) sequenced. A printout of the actual sequence data and the
identification is included in each report.
Vidas™ (Vitek Immuno Diagnostic System)
The VIDAS provides a more automated method of running ELISA-like assays
for the qualitative detection of specific bacterial pathogens. We can
run up to 30 samples in five separate bays all at one time or each aby
on its own schedule with up to six samples. It uses a fluorometric
detector to read results, instead of the typically less sensitive
colorimetric detection seen in other assay methods. Currently, we
employ the VIDAS instrument for Salmonell, Listeria and E.coli O157:H7 assays.
Vitek® 2 Compact
The VITEK® 2 Anaerobic and Coryenbacteria identification card (ANC) is
intended for use with VITEK® 2 Systems for the automated identification
of most clinically significant anaerobic organisms and Corynebacterium
species. The ANC card is based on established biochemical methods and
newly developed substrates. There are 36 biochemical tests measuring
carbon source utilization and enzymatic activities. Final results are
available in approximately six hours.
We are able to identify an organism by using a methodology based on
the characteristics of the data & knowledge about the organism and
reactions being analyzed. Sufficient data have been collected from
known strains to estimate the typical reactions of the claimed species
to a set of discriminating biochemicals. If a unique identification
pattern is not recognized, a list of possible organisms is given, or
the strain is determined to be outside the scope of the databases.
GC-FAME + Biolog®
Gram negative Bacteria
Gram Positive Bacteria
26S LSU Fungal Analysis
Mold/Fungal GC-FAME Analysis
Yeast
For further information about methods used, contact us.
