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GeneQuence™
Novel Rapid Pathogen Detection – Sensitive, Simple, and
Scalable - AOAC Official Method Approved
Protocol Overview:
First, a portion of the enrichment culture is placed into a test
tube. A lysis reagent is added, which disrupts the cell and
releases the nucleic acid target molecules. A portion of the
lysed sample is then transferred to a microwell and the probe
reagents are added. (In the GENE-TRAK® DLP format, the
probe reagents are added to the tube with the lysed sample,
followed by introduction of the coated dipstick to the tube.)
Point of Benefit of
Comparison GeneQuence PCR GeneQuence
Assay time 1 hr, 50 min 3 hrs, 30 Faster results.
min GeneQuence also
(average) has at least 25%
less off-line labor
time (instrument
set-up, manual
reagent addition,
etc.) than PCR
methods.
Commodities Good Some Published data
performance food documents
across foods types problems PCR in
difficult certain food
commodities.
Flexibility 4 pathogens 1 GeneQuence, due
at same time pathogen to its superior
per run automation,
allows 4
pathogens to be
assayed
concurrently. To
do this by PCR,
more than one
instrument would
be needed (at a
higher capital
cost).
Processing 4 96-well 1 set of As soon as one
plates 96 plate is finished,
concurrently samples GeneQuence can
at a time accept another
sample set. PCR
systems must wait
until the entire run
is complete before
loading another
set. GeneQuence
can provide
uninterrupted,
continuous
operation.
Technology Nucleic acid PCR The thermostable
hybridization polymerase used
in PCR has
documented
inhibition
problems that
lead to false
negatives with
several matrices.
GeneQuence
avoids this issue
entirely by using
nucleic acid
hybridization.
GeneQuence also
probes the target
directly, rather
than an
intermediate
amplified from the
original target.
Throughput 1.32 samples 0.46 187% more
per minute samples throughput on
per GeneQuence will
minute minimize retention
time of inventory.
References:
Optimal polymerase varies per food type
Abu Al-Soud, W. and Radstrom, P. 1998. Appl. Environ.
Microbiol. 64: 3748-3753.
Point of Benefit of
Comparison GeneQuence ELISA GeneQuence
Detection rRNA Protein 1,000 to 10,000
copies of rRNA per
cell give
GeneQuence its
high level of
sensitivity.
Inclusivity DNA Sandwich Allows detection of
Hybridization ELISA all serovars of
Salmonella
enteritidis
Pathogen rRNA Protein RNA is very
Target short-lived as it is
constantly broken
down by ubiquitous
RNases. Couple
that with dilutions
from enrichment,
GeneQuence will
not detect dead
bacteria.
Procedure Manual or Varies Some ELISA
Automated systems can only
be run on an
instrument. Others
specify only a
manual method that
may be adapted by
the user into an
automated system.
Only GeneQuence
allows the user to
perform the test
manually or in a
company-supported
automated format.
Specificity DNA:rRNA Antibody: Two levels of
Protein stringency (binding
of target rRNA to
capture probe, and
rRNA to detector
probe) decrease
false positive rates
with GeneQuence.
GENE-TRAK® Assays
GENE-TRAK is the classic version of our GeneQuence
technology, where instead of being performed in a microtiter
plate, the assay utilized small ‘dipstick’ paddles to which the
capture probe is bound. The DLP (direct labeled probe)
employs specific DNA probes which are directly labeled with
horseradish peroxidase, thereby decreasing the time by ~20
minutes.
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Neogen Corporation
800/234-5333 (USA/Canada) or 517/372-9200
Fax: 517/372-2006 • foodsafety@neogen.com