Genequence Automated System
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Technology Overview | Advantages | Fully Automated System | Rapid Pathogen Kits Web Seminar Request | Technical Resources | Request More Information | GENE-TRAK Assays Food Safety Main Page | Foodborne Bacteria
GeneQuence™ Novel Rapid Pathogen Detection – Sensitive, Simple, and Scalable - AOAC Official Method Approved GeneQuence utilizes a novel DNA hybridization technology which assays for Salmonella, Listeria spp., Listeria monocytogenes, and E.coli O157:H7* Each test kit uses two specific DNA elements ensuring the highest of specificity, thereby increasing your confidence of the rapid results. This web portal is designed to provide you with all of the technical and product related information on the GeneQuence and GENE-TRAK rapid pathogen detection platforms. Each kit has been developed to detect 1-5 CFU/25g sample in less than 2 hours. By coupling this easy to use technology with an automated plate handling unit , it is possible test more than 700 samples in an 8 hour work day with very little hands on time. Contact us if you are ready to simplify your pathogen testing program. Technology that performs GeneQuence is based upon the DNA-probe technology that lead to broad adoption of the classic GENE-TRAK Pathogen Assays. Each test kit contains capture and detector DNA probes specific to ribosomal RNA (rRNA) of the target organism and a coated solid phase (dipstick or microwell). To achieve a positive result both the capture probe AND the detector probe must bind. This dual selectivity ensures that only the organism of interest is detected during the assay. 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.)
The probe reagents consist of: 1) an oligonucleotide capture probe specific to rRNA sequences of the target organism and labeled at the 3’ end with polydeoxyadenylic acid (poly dA); and 2) an oligonucleotide detector probe also specific to rRNA sequences of the target organism and labeled at the 5’
22/02/2007 11:12 a.m.
GeneQuence Automated System
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http://www.neogen.com/genequence.htm
end with the enzyme horseradish peroxidase (HRP). The hybridization reaction is then allowed to proceed for one hour. If target rRNA is present in the sample, both probes will hybridize to their complementary sequences on the target molecule.
The resulting complex is captured onto the solid phase coated with polydeoxythymidylic acid (poly dT), which is complementary to the poly dA portion of the capture probe. Unbound probe is then washed away, and a substrate of HRP is added.
Following a short incubation, blue color indicates the presence of hybridized detector probe in the complex and thus the presence of rRNA from the target organism. Stop solution is added to convert blue to yellow color and results are determined spectrophotometrically at 450 nm. An absorbance value in excess of a predetermined threshold indicates a positive test result.
The GeneQuence Advantage GeneQuence DNA Hybridization Method versus PCR-based Methods Point of Comparison GeneQuence PCR Assay time
1 hr, 50 min
Benefit of GeneQuence
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 performance across foods
Some food types difficult
Flexibility
1 GeneQuence, due pathogen to its superior per run automation, allows 4 pathogens to be assayed
4 pathogens at same time
Published data documents problems PCR in certain food commodities.
22/02/2007 11:12 a.m.
GeneQuence Automated System
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http://www.neogen.com/genequence.htm
concurrently. To do this by PCR, more than one instrument would be needed (at a higher capital cost). Processing
4 96-well plates concurrently
1 set of 96 samples at a time
As soon as one plate is finished, GeneQuence can 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 hybridization
PCR
The thermostable 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 per minute
0.46 samples per minute
187% more throughput on GeneQuence will 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. High false negatives (vs. ELISA and semi-solid medium) Uyttendaele, M. et al. 2003. Lett. Appl. Microbiol. 37: 386-391. Biological samples affect sensitivity and kinetics of PCR Radstrom, P. et al. 2004. Mol. Biotechnol. 26: 133-146. Lower sensitivity (than ELISA) in seafood
22/02/2007 11:12 a.m.
GeneQuence Automated System
4 de 5
http://www.neogen.com/genequence.htm
Thompson, S. et al. 2003. ASM General Meeting Abstracts. Evaluation of PCR versus culture based detection of E.coli 0157:H7 in Ground Beef Arthur, T.M. et al. 2005. J. of Food Protect. 68(8): 1566-1574.
GeneQuence DNA Hybridization Method versus Antibody-Microwell Based Methods Point of Comparison GeneQuence ELISA
Benefit of GeneQuence
Detection
rRNA
Protein
1,000 to 10,000 copies of rRNA per cell give GeneQuence its high level of sensitivity.
Inclusivity
DNA Hybridization
Sandwich Allows detection of ELISA all serovars of Salmonella enteritidis
Pathogen Target
rRNA
Protein
RNA is very 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 Automated
Varies
Some ELISA 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.
22/02/2007 11:12 a.m.
GeneQuence Automated System
5 de 5
http://www.neogen.com/genequence.htm
Throughput
DSX
Other The DSX can platforms process more than 744 samples in an 8-hour shift, at least double that of any ELISA system on the market. This will reduce labor and equipment costs.
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. GENE-TRAK demonstrates the same high specificity found in our GeneQuence microwell assay but is perfect for those who test less than 15 samples per evaluation. GENE-TRAK is highly referenced in the industry and has additional target organisms such as Staphylococcus aureus, Campylobacter spp. and E.coli (contact food safety customer service for more information) GENE-TRAK pathogen detection products: GENE-TRAK Listeria Assay GENE-TRAK Listeria DLP Assay GENE-TRAK Listeria monocytogenes Assay GENE-TRAK Salmonella Assay GENE-TRAK Salmonella DLP Assay GENE-TRAK Staphylococcus aureus Assay GENE-TRAK E. coli Assay Return to top View GeneQuence Assay Kits | Food Safety Main Page | Foodborne Bacteria
Neogen Corporation 800/234-5333 (USA/Canada) or 517/372-9200 Fax: 517/372-2006 • [email protected]
Neogen Home | Food Safety | Animal Safety | Life Sciences | Acumedia Profile | Press | Investors | International | Ordering | Employment | Other Links ©Neogen Corporation
22/02/2007 11:12 a.m.
1 de 5
http://www.neogen.com/genequence.htm
Technology Overview | Advantages | Fully Automated System | Rapid Pathogen Kits Web Seminar Request | Technical Resources | Request More Information | GENE-TRAK Assays Food Safety Main Page | Foodborne Bacteria
GeneQuence™ Novel Rapid Pathogen Detection – Sensitive, Simple, and Scalable - AOAC Official Method Approved GeneQuence utilizes a novel DNA hybridization technology which assays for Salmonella, Listeria spp., Listeria monocytogenes, and E.coli O157:H7* Each test kit uses two specific DNA elements ensuring the highest of specificity, thereby increasing your confidence of the rapid results. This web portal is designed to provide you with all of the technical and product related information on the GeneQuence and GENE-TRAK rapid pathogen detection platforms. Each kit has been developed to detect 1-5 CFU/25g sample in less than 2 hours. By coupling this easy to use technology with an automated plate handling unit , it is possible test more than 700 samples in an 8 hour work day with very little hands on time. Contact us if you are ready to simplify your pathogen testing program. Technology that performs GeneQuence is based upon the DNA-probe technology that lead to broad adoption of the classic GENE-TRAK Pathogen Assays. Each test kit contains capture and detector DNA probes specific to ribosomal RNA (rRNA) of the target organism and a coated solid phase (dipstick or microwell). To achieve a positive result both the capture probe AND the detector probe must bind. This dual selectivity ensures that only the organism of interest is detected during the assay. 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.)
The probe reagents consist of: 1) an oligonucleotide capture probe specific to rRNA sequences of the target organism and labeled at the 3’ end with polydeoxyadenylic acid (poly dA); and 2) an oligonucleotide detector probe also specific to rRNA sequences of the target organism and labeled at the 5’
22/02/2007 11:12 a.m.
GeneQuence Automated System
2 de 5
http://www.neogen.com/genequence.htm
end with the enzyme horseradish peroxidase (HRP). The hybridization reaction is then allowed to proceed for one hour. If target rRNA is present in the sample, both probes will hybridize to their complementary sequences on the target molecule.
The resulting complex is captured onto the solid phase coated with polydeoxythymidylic acid (poly dT), which is complementary to the poly dA portion of the capture probe. Unbound probe is then washed away, and a substrate of HRP is added.
Following a short incubation, blue color indicates the presence of hybridized detector probe in the complex and thus the presence of rRNA from the target organism. Stop solution is added to convert blue to yellow color and results are determined spectrophotometrically at 450 nm. An absorbance value in excess of a predetermined threshold indicates a positive test result.
The GeneQuence Advantage GeneQuence DNA Hybridization Method versus PCR-based Methods Point of Comparison GeneQuence PCR Assay time
1 hr, 50 min
Benefit of GeneQuence
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 performance across foods
Some food types difficult
Flexibility
1 GeneQuence, due pathogen to its superior per run automation, allows 4 pathogens to be assayed
4 pathogens at same time
Published data documents problems PCR in certain food commodities.
22/02/2007 11:12 a.m.
GeneQuence Automated System
3 de 5
http://www.neogen.com/genequence.htm
concurrently. To do this by PCR, more than one instrument would be needed (at a higher capital cost). Processing
4 96-well plates concurrently
1 set of 96 samples at a time
As soon as one plate is finished, GeneQuence can 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 hybridization
PCR
The thermostable 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 per minute
0.46 samples per minute
187% more throughput on GeneQuence will 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. High false negatives (vs. ELISA and semi-solid medium) Uyttendaele, M. et al. 2003. Lett. Appl. Microbiol. 37: 386-391. Biological samples affect sensitivity and kinetics of PCR Radstrom, P. et al. 2004. Mol. Biotechnol. 26: 133-146. Lower sensitivity (than ELISA) in seafood
22/02/2007 11:12 a.m.
GeneQuence Automated System
4 de 5
http://www.neogen.com/genequence.htm
Thompson, S. et al. 2003. ASM General Meeting Abstracts. Evaluation of PCR versus culture based detection of E.coli 0157:H7 in Ground Beef Arthur, T.M. et al. 2005. J. of Food Protect. 68(8): 1566-1574.
GeneQuence DNA Hybridization Method versus Antibody-Microwell Based Methods Point of Comparison GeneQuence ELISA
Benefit of GeneQuence
Detection
rRNA
Protein
1,000 to 10,000 copies of rRNA per cell give GeneQuence its high level of sensitivity.
Inclusivity
DNA Hybridization
Sandwich Allows detection of ELISA all serovars of Salmonella enteritidis
Pathogen Target
rRNA
Protein
RNA is very 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 Automated
Varies
Some ELISA 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.
22/02/2007 11:12 a.m.
GeneQuence Automated System
5 de 5
http://www.neogen.com/genequence.htm
Throughput
DSX
Other The DSX can platforms process more than 744 samples in an 8-hour shift, at least double that of any ELISA system on the market. This will reduce labor and equipment costs.
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. GENE-TRAK demonstrates the same high specificity found in our GeneQuence microwell assay but is perfect for those who test less than 15 samples per evaluation. GENE-TRAK is highly referenced in the industry and has additional target organisms such as Staphylococcus aureus, Campylobacter spp. and E.coli (contact food safety customer service for more information) GENE-TRAK pathogen detection products: GENE-TRAK Listeria Assay GENE-TRAK Listeria DLP Assay GENE-TRAK Listeria monocytogenes Assay GENE-TRAK Salmonella Assay GENE-TRAK Salmonella DLP Assay GENE-TRAK Staphylococcus aureus Assay GENE-TRAK E. coli Assay Return to top View GeneQuence Assay Kits | Food Safety Main Page | Foodborne Bacteria
Neogen Corporation 800/234-5333 (USA/Canada) or 517/372-9200 Fax: 517/372-2006 • [email protected]
Neogen Home | Food Safety | Animal Safety | Life Sciences | Acumedia Profile | Press | Investors | International | Ordering | Employment | Other Links ©Neogen Corporation
22/02/2007 11:12 a.m.
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