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RapidFinder™ Quant Multi-Meat Set

The RapidFinder™ Quant Multi-Meat Set, when used with one or more of the RapidFinder™ Meat ID kits listed below, enables the determination of the percentage of a target meat species with respect to total meat in food and feed samples. Using the included standards, the kit enables quantification of as little as 0.05% target species DNA as compared to other meat DNA in only a few hours.

The RapidFinder Quant Multi-Meat Set is designed to be used in combination with one or more of the following meat species-specific ID kits (available separately):

NameCatalog Number
RapidFinder™ Beef ID KitA24391
RapidFinder™ Pork ID KitA24392
RapidFinder™ Equine ID KitA15570
RapidFinder™ Poultry ID KitA24397
RapidFinder™ Chicken ID KitA24393
RapidFinder™ Turkey ID KitA24394


• Simple-real-time PCR based assay, no electrophoresis needed
• Flexible-compatible with any food or feed sample, both raw and processed
• Reliable-Includes multi-meat DNA quantification standard
• Fast-from sample to result in the same day

How It Works
The method relies on quantitative real-time PCR. First, a standard curve is created by serially diluting the included multi-meat DNA standard. Then assays for the target species DNA (from one of the kits listed above) and for all meat DNA (from the RapidFinder™ Quant Multi-Meat Set) are run side-by-side on both the food sample DNA and the DNA standards. This approach enables rapid and accurate quantification and reporting of the percentage of target meat species as compared to total meat in a sample. Included in each reaction mix is an internal positive control detected in the VIC® dye channel, which is used to verify the absence of PCR inhibitors.

The RapidFinder™ Quant Multi-Meat Set is compatible with Applied Biosystems® StepOne™, StepOnePlus™, 7500, 7500 Fast, and 7300 real-time PCR systems.

Sample Preparation
DNA sample preparation for the RapidFinder™ meat species identification and quantification workflow is accomplished with the easy-to-use GMO Extraction Kit (Cat. No. 4466336). Using 10 g of sample rather than the smaller sample sizes accommodated by most other DNA isolation procedures minimizes sampling errors. The GMO Extraction Kit is compatible with raw and processed meat, as well as composite food samples.

RapidFinder™ Beef ID Kit

The RapidFinder™ Beef ID Kit enables the simple and reliable detection of beef in food and feed samples. The method works by detection of bovine, or Bos taurus, DNA by real-time PCR. Unlike ELISA-based methods, when used with the GMO Extraction Kit (Cat. No. 4466336) for sample preparation, the RapidFinder™ Beef ID Kit detects both raw and processed beef and provides sensitivity down to 0.01% bovine DNA. The included positive control, which comprises 0.1% bovine DNA, can be used as a reference to set the threshold of detection of the assay.

• Simple-real-time PCR based assay, no electrophoresis needed
• Flexible-bovine DNA detected in any food or feed sample, both raw and processed
• Reliable-Internal Positive Control (IPC) provides process-control check for PCR inhibitors
• Fast-from sample to result in the same day

How the RapidFinder™ Beef ID Kit Works
Bovine DNA detection is accomplished by real-time PCR using the supplied primer/probe set specific for mitochondrial DNA sequence specific to Bos taurus. The target sequence, detected in the FAM™ dye channel, is not only specific for beef, but also stable. Also included in each reaction mix is an internal positive control detected in the VIC® dye channel, which is used to verify the absence of PCR inhibitors.

By performing a separate reaction with the included positive control, users are able to verify the system and assay are working correctly. More importantly, it enables easy data interpretation. The positive control is provided as 0.1% bovine DNA in a background of fish DNA, and by comparing sample results to the control results, a threshold of detection is provided.

The assay is compatible with Applied Biosystems® StepOne™, StepOnePlus™, 7500, 7500 Fast, and 7300 real-time PCR systems.

Sample Preparation
DNA sample preparation for the RapidFinder™ Beef ID Kit workflow is accomplished with the easy-to-use GMO Extraction Kit (Cat. No. 4466336). Using 10 g of sample rather than the smaller sample sizes accommodated by most other DNA isolation procedures minimizes sampling errors. The GMO Extraction Kit is compatible with raw and processed meat, as well as composite food samples.

Quantification
If a positive result is obtained with the RapidFinder™ Beef ID Kit, most labs will go on to quantify the amount of bovine DNA in the sample. This can be accomplished using the RapidFinder™ Beef ID Kit in conjunction with the RapidFinder™ Quant Multi-Meat Set (Cat. No. A24399).

Part of a Suite of Solutions For Meat Species Detection
The RapidFinder™ Beef ID Kit is just one of our many kits available for the sensitive detection of various meat species in food and feed samples. Other RapidFinder™ meat species identification kits available include those for the detection of pork, horse, sheep, ruminant, chicken, turkey, and poultry.

The RapidFinder™ Meat Species Identification kits are made available through a partnership with IMEGEN (Instituto de Medicina Genomica).

mirVana™ miRNA Detection Kit (Invitrogen™)

The Ambion® mirVana™ miRNA Detection Kit provides a fast and sensitive method for detecting small RNAs. The assay is 100–500 times more sensitive than Northern analysis, it is able to detect as little as 10 attomoles (10-17 mol) of target RNA. Each kit includes sufficient reagents for 100 reactions.

• Sensitive—detect miRNA or siRNA in as little as 50 ng total RNA
• Specific—extremely low backgrounds
• Simple and fast—single-tube procedure eliminates the need to transfer to membrane for hybridization
• Multiple target detection—detect multiple small RNAs and mRNAs in the same sample

Rapid Procedure Based on Solution Hybridization
The mirVana™ miRNA Detection Kit is based on a simple solution hybridization principle where the RNA sample is simply mixed with radiolabeled RNA probe(s) and hybridized. Unhybridized RNA and excess probe are removed by a digestion step and the hybridized, protected RNA fragments are then recovered using a patented single-step technology for simultaneous ribonuclease inactivation and nucleic acid precipitation. RNA samples are then resuspended and analyzed on a denaturing polyacrylamide gel.

Simultaneous Detection of siRNA Expression and Target Gene Knockdown
Because multiple RNA species of the same or differing size can be detected in the same sample, the mirVana™ miRNA Detection Kit is ideal for correlating siRNA expression levels with target mRNA knockdown. An example of this application is shown in the accompanying data, which demonstrates that GAPDH mRNA levels were reduced in cells expressing a GAPDH siRNA, but not in cells expressing a control siRNA.

Accessory Products:
The mirVana™ miRNA Probe Construction Kit (SKU#AM1550) is available for the preparation and labeling of probes by in vitro transcription and the mirVana™ Probe & Marker Kit is available for 5' end labeling of oligonucleotide probes and the included Decade Markers (SKU#AM1554).

RapidFinder™ Pork ID Kit

The RapidFinder™ Pork ID Kit enables the simple and reliable detection of pork in food and feed samples. The method works by detection of porcine, or Sus scrofa, DNA by real-time PCR. Unlike ELISA based methods, when used with the GMO Extraction Kit (Cat. No. 4466336) for sample preparation, the RapidFinder™ Pork ID Kit detects both raw and processed pork and provides sensitivity down to 0.01% porcine DNA. The included positive control, which comprises 0.1% porcine DNA, can be used as a reference to set the threshold of detection of the assay.

• Simple-real-time PCR based assay, no electrophoresis needed
• Flexible-porcine DNA detected in any food or feed sample, both raw and processed
• Reliable-Internal Positive Control (IPC) provides process control check for PCR inhibitors
• Fast-from sample to result in the same day

How the RapidFinder™ Pork ID Kit Works
Porcine DNA detection is accomplished by real-time PCR using the supplied primer/probe set specific for mitochondrial DNA sequence specific to Sus scrofa (including Sus scrofa domesticus, commonly referred to as Sus domesticus). The target sequence, detected in the FAM™ dye channel, is not only specific for pork, but also stable. Also included in each reaction mix is an internal positive control detected in the VIC® dye channel, which is used to verify the absence of PCR inhibitors.

By performing a separate reaction with the included positive control, users are able to verify the system and assay are working correctly. More importantly, it enables easy data interpretation. The positive control is provided as 0.1% porcine DNA in a background of fish DNA, and by comparing sample results to the control results, a threshold of detection is provided.

The assay is compatible with Applied Biosystems® StepOne™, StepOnePlus™, 7500, 7500 Fast, and 7300 real-time PCR systems.

Sample Preparation
DNA sample preparation for the RapidFinder™ Pork ID Kit workflow is accomplished with the easy-to-use GMO Extraction Kit (Cat. No. 4466336). Using 10 g of sample rather than the smaller sample sizes accommodated by most other DNA isolation procedures minimizes sampling errors. The GMO Extraction Kit is compatible with raw and processed meat, as well as composite food samples.

Quantification
If a positive result is obtained with the RapidFinder™ Pork ID Kit, most labs will go on to quantify the amount of porcine DNA in the sample. This can be accomplished using the RapidFinder™ Pork ID Kit in conjunction with the RapidFinder™ Quant Multi-Meat Set (Cat. No. A24399).

Part of a Suite of Solutions For Meat Species Detection
The RapidFinder™ Pork ID Kit is just one of our many available kits for the sensitive detection of various meat species in food and feed samples. Other RapidFinder™ meat species identification kits available include those for the detection of beef, horse, sheep, ruminant, chicken, turkey, and poultry.

The RapidFinder™ Meat Species Identification kits are made available through a partnership with IMEGEN (Instituto de Medicina Genomica).

ViewRNA™ Cell Plus Cytospin Module Kit (Invitrogen™)

The ViewRNA™ Cell Plus Cytospin Module Kit is for use with the ViewRNA™ Cell Plus Assay kit (cat. 88-19000). These buffers can be used to perform the assay when using cytospin slides with cells.

Please refer to the ViewRNA™ Cell Plus Assay protocol on the product data sheet for instructions for use.

Reported Application
Microscopy, Immunocytochemistry

RapidFinder™ Chicken ID Kit

The RapidFinder™ Chicken ID Kit enables the simple and reliable detection of chicken in food and feed samples. The method works by detection of chicken, or Gallus gallus, DNA by real-time PCR. Unlike ELISA based methods, when used with the GMO Extraction Kit (Cat. No. 4466336) for sample preparation, the RapidFinder™ Chicken ID Kit detects both raw and processed chicken and provides sensitivity down to 0.01% chicken DNA. The included positive control, which comprises 0.1% chicken DNA, can be used as a reference to set the threshold of detection of the assay.

• Simple-real-time PCR based assay, no electrophoresis needed
• Flexible-chicken DNA detected in any food or feed sample, both raw and processed
• Reliable-Internal Positive Control (IPC) provides process control check for PCR inhibitors
• Fast-from sample to result in the same day

How the RapidFinder™ Chicken ID Kit Works
Chicken DNA detection is accomplished by real-time PCR using the supplied primer/probe set specific for mitochondrial DNA sequence specific to Gallus gallus. The target sequence, detected in the FAM™ dye channel, is not only specific for chicken, but also stable. Also included in each reaction mix is an internal positive control detected in the VIC® dye channel, which is used to verify the absence of PCR inhibitors.

By performing a separate reaction with the included positive control, users are able to verify the system and assay are working correctly. More importantly, it enables easy data interpretation. The positive control is provided as 0.1% chicken DNA in a background of fish DNA, and by comparing sample results to the control results, a threshold of detection is provided.

The assay is compatible with Applied Biosystems® StepOne™, StepOnePlus™, 7500, 7500 Fast, and 7300 real-time PCR systems.

Sample Preparation
DNA sample preparation for the RapidFinder™ Chicken ID Kit workflow is accomplished with the easy-to-use GMO Extraction Kit (Cat. No. 4466336). Using 10 g of sample rather than the smaller sample sizes accommodated by most other DNA isolation procedures minimizes sampling errors. The GMO Extraction Kit is compatible with raw and processed meat, as well as composite food samples.

Quantification
If a positive result is obtained with the RapidFinder™ Chicken ID Kit, most labs will go on to quantify the amount of chicken DNA in the sample. This can be accomplished using the RapidFinder™ Chicken ID Kit in conjunction with the RapidFinder™ Quant Multi-Meat Set (Cat. No. A24399).

Part of a Suite of Solutions For Meat Species Detection
The RapidFinder™ Chicken ID Kit is just one of our many kits available for the sensitive detection of various meat species in food and feed samples. Other RapidFinder™ meat species identification kits available include those for the detection of pork, beef, horse, sheep, ruminant, turkey, and poultry.

The RapidFinder™ Meat Species Identification Kits are made available through a partnership with IMEGEN (Instituto de Medicina Genomica).

RapidFinder™ Ruminant ID Kit

The RapidFinder™ Ruminant ID Kit enables the simple and reliable detection of ruminant meat in food and feed samples. The method works by detection of Bos taurus, Ovis aries, Capra hircus, Cervus elaphus, and Capreolus capreolus DNA, which corresponds to DNA from beef, sheep, goat, and deer, by real-time PCR. Unlike ELISA based methods, when used with the GMO Extraction Kit (Cat. No. 4466336) for sample preparation, the RapidFinder™ Ruminant ID Kit detects both raw and processed ruminant meat and provides sensitivity down to 0.01% ruminant DNA. The included positive control in the kit, which comprises 0.1% ruminant DNA, can be used as a reference to set the threshold of detection of the assay.

• Simple-real-time PCR based assay, no electrophoresis needed
• Flexible-ruminant DNA detected in any food or feed sample, both raw and processed
• Reliable-Internal Positive Control (IPC) provides process control check for PCR inhibitors
• Fast-from sample to result in the same day

How the RapidFinder™ Ruminant ID Kit Works
Ruminant DNA detection is accomplished by real-time PCR using the supplied primer/probe mixes targeting a multi-copy nuclear target specific to Bos taurus, Ovis aries, Capra hircus, Cervus elaphus, and Capreolus capreolus. The primer/probes are supplied in a single tube, so if any of the five included ruminant species is detected, a positive result will be obtained. The target sequences, detected in the FAM dye channel, are not only specific for the five ruminants, but are also stable. Also included in each reaction mix is an internal positive control detected in the VIC dye channel, which is used to verify the absence of PCR inhibitors.

By performing a separate reaction with the included positive control, users are able to verify the system and assay are working correctly. More importantly, however, it enables easy data interpretation. The positive control is provided as 0.1% Bos taurus DNA in a background of fish DNA, and by comparing sample results to the control results, a threshold of detection is provided.

The assay is compatible with Applied Biosystems® StepOne™, StepOnePlus™, 7500, 7500 Fast, and 7300 real-time PCR systems.

Sample Preparation
DNA sample preparation for the RapidFinder™ Ruminant ID Kit workflow is accomplished with the easy-to-use GMO Extraction Kit (Cat. No. 4466336). Using 10 g of sample rather than the smaller sample sizes accommodated by most other DNA isolation procedures minimizes sampling errors. The GMO Extraction Kit is compatible with raw and processed meat, as well as composite food samples.

Part of a Suite of Solutions For Meat Species Detection
The RapidFinder™ Ruminant ID Kit is just one of our many available kits for the sensitive detection of various meat species in food and feed samples. Other RapidFinder™ meat species identification kits available include those for the detection of pork, beef, horse, sheep, chicken, turkey, and poultry.

The RapidFinder™ Meat Species Identification Kits are made available through a partnership with IMEGEN (Instituto de Medicina Genomica).

RapidFinder™ STEC Confirmation Assay (Applied Biosystems™)

The Applied Biosystems™ RapidFinder™ STEC Confirmation Assay is part of a complete, validated screening and confirmation workflow enabling TaqMan™ real-time PCR detection of the Shiga toxin-producing E. coli (STEC) O157:H7 and Big 6 non‐O157:H7 serogroup (O26, O45, O103, O111, O121, and O145) in a variety of beef products. Samples assessed as positive using the RapidFinder STEC Screening Assay are further tested using this RapidFinder STEC Confirmation Assay. All reactions include an internal positive control (IPC) and ROX™ passive reference.

The RapidFinder STEC Detection Workflow provides:
• Greater confidence in results—part of a validated workflow for detection of as little as 1-5 CFU of pathogenic STEC in your sample
• Simplified workflow—simultaneous detection of E. coli O157:H7 and the Big 6 non-O157:H7 strains in 375 g beef samples with a closed-tube, single-step lyophilized detection format that fits within your existing testing program
• Reduced chance of false results—through use of high-quality DNA preparation, proprietary assay design, and internal positive control
• Same day detection of pathogenic STECs and clearance of STEC-negative beef samples in as little as 12 hours, supporting early product release
• Minimized costs and laboratory space with the use a lower volume of a nonproprietary single-step enrichment medium, eliminating the need to purchase proprietary media
• Compatibility with RapidFinder™ Express software v.1.2 and later—enables easy, automated presence/absence detection and interpretation of results

Studies have shown that the RapidFinder STEC assays have a high level of sensitivity and specificity for E. coli O157:H7 and the Big 6 non-O157 STEC strains*. They detected all inclusion strains and showed no cross-reactivity to any of the exclusion strains tested. The stx assays detected all known variants of stx1 and stx2, including stx2f and stx2g. The optimized workflow co-developed with USDA-ARS showed equivalent detection to the MLG reference method, and the internal positive control in both assays significantly reduces the risk of false-negatives.

Applications
The RapidFinder STEC Detection Workflow is intended for use by microbiological analysts who need to test for STEC in beef samples. Combined with the Applied Biosystems 7500 Fast RT-PCR System, RapidFinder STEC screening and confirmation assays provide a comprehensive solution for rapid detection, identification, and confirmation of Big 6 STECs and E. coli 0157:H7 to meet importation and exportation requirements for the beef industry. Specific workflows are detailed in the figures below and include both automated and spin column-based DNA isolation and real-time PCR detection of E. coli O157:H7 and Big 6 non-O157 strains in beef samples.

RapidFinder Express Software
When integrated into the RapidFinder STEC Detection Workflow, RapidFinder Express Software automates data interpretation and generates user instructions on how to proceed with a sample. Data displays are available for novice and advanced users, showing simple presence/absence calls or raw data files using 7500 Fast System SDS Software.

RapidFinder STEC Screening and Confirmation Assays meet USDA FSIS guidance
The RapidFinder STEC Detection Workflow, a two-stage real-time PCR method, is designed to accurately detect, identify, and confirm E. coli O157:H7 and the Big 6 serogroups of non-O157 Shiga toxin-producing E. coli defined by the USDA Food Safety and Inspection Service (FSIS) as adulterants in the American beef industry.

Learn more about the RapidFinder STEC Detection Workflow >

*Swimley, Michelle S. Design and Evaluation of Two-Stage Multiplex Real-Time PCR Method for Detecting O157:H7 and non-O157 STEC Strains from Beef Samples. IAFP 2015.

RapidFinder™ Turkey ID Kit

The RapidFinder™ Turkey ID Kit enables the simple and reliable detection of turkey in food and feed samples. The method works by detection of turkey, or Meleagris gallopavo, DNA by real-time PCR. Unlike ELISA based methods, when used with the GMO Extraction Kit (Cat. No. 4466336) for sample preparation, the RapidFinder™ Turkey ID Kit detects both raw and processed turkey and provides sensitivity down to 0.01% turkey DNA. The included positive control in the kit, which comprises 0.1% turkey DNA, can be used as a reference to set the threshold of detection of the assay.

• Simple-real-time PCR based assay, no electrophoresis needed
• Flexible-turkey DNA detected in any food or feed sample, both raw and processed
• Reliable-Internal Positive Control (IPC) provides process control check for PCR inhibitors
• Fast-from sample to result in the same day

How the RapidFinder™ Turkey ID Kit Works
Turkey DNA detection is accomplished by real-time PCR using the supplied primer/probe set specific for mitochondrial DNA sequence specific to Meleagris gallopavo. The target sequence, detected in the FAM™ dye channel, is not only specific for turkey, but also stable. Also included in each reaction mix is an internal positive control detected in the VIC® dye channel, which is used to verify the absence of PCR inhibitors.

By performing a separate reaction with the included positive control, users are able to verify the system and assay are working correctly. More importantly, it enables easy data interpretation. The positive control is provided as 0.1% turkey DNA in a background of fish DNA, and by comparing sample results to the control results, a threshold of detection is provided.

The assay is compatible with Applied Biosystems® StepOne™, StepOnePlus™, 7500, 7500 Fast, and 7300 real-time PCR systems.

Sample Preparation
DNA sample preparation for the RapidFinder™ Turkey ID Kit workflow is accomplished with the easy-to-use GMO Extraction Kit (Cat. No. 4466336). Using 10 g of sample rather than smaller sample sizes accommodated by most other DNA isolation procedures minimizes sampling errors. The GMO Extraction Kit is compatible with raw and processed meat, as well as composite food samples.

Quantification
If a positive result is obtained with the RapidFinder™ Turkey ID Kit, most labs will go on to quantify the amount of turkey DNA in the sample. This can be accomplished using the RapidFinder™ Turkey ID Kit in conjunction with the RapidFinder™ Quant Multi-Meat Set (Cat. No. A24399).

Part of a Suite of Solutions For Meat Species Detection
The RapidFinder™ Turkey ID Kit is just one of our many available kits for the sensitive detection of various meat species in food and feed samples. Other RapidFinder™ meat species identification kits available include those for the detection of pork, beef, horse, sheep, ruminant, chicken, and poultry.

The RapidFinder™ Meat Species Identification Kits are made available through a partnership with IMEGEN (Instituto de Medicina Genomica).

PrimeFlow™ RNA Assay Kit (Invitrogen™)

This PrimeFlow™ RNA Assay is an in situ hybridization assay that combines the power of branched-DNA technology with the single-cell resolution of flow cytometry, enabling the simultaneous detection of up to four RNA targets in combination with immunophenotyping for cell surface and intracellular proteins using fluorochrome-conjugated antibodies. Branched-DNA technology uses pairs of target-specific probes to amplify the detection of a RNA transcript, achieving between 8,000- to 16,000-fold signal amplification from a desired RNA transcript. PrimeFlow RNA Assay can detect mRNA, long non-coding RNA, viral RNA, and telomere DNA. The detection of microRNA is also possible and is optimal when using the PrimeFlow™ microRNA Pretreatment Buffer (cat. 88-18006) and protocol. PrimeFlow RNA Assay is compatible with human and mouse cells. Other mammalian and non-mammalian cells may be compatible; please contact LuminexFAS@thermofisher.com for more information.

This PrimeFlow RNA Assay contains all the reagents needed to conduct the assay. Positive control probes and target probes for genes of interest are sold separately.



Reported Application
RNA Flow Cytometry

RapidFinder™ STEC Screening Assay (Applied Biosystems™)

The Applied Biosystems™ RapidFinder™ STEC Screening Assay is part of a complete, validated screening and confirmation workflow enabling TaqMan™ real-time PCR detection of the Shiga toxin-producing E. coli (STEC) O157:H7 and Big 6 non‐O157:H7 serogroup (O26, O45, O103, O111, O121 and O145) in a variety of beef products. The RapidFinder STEC Screening Assay was co-developed with the USDA-ARS and contains 96 complete qPCR assays targeted to screen for Shiga toxin 1 and 2 (stx1/stx2), eae, and potential O157:H7 presence. The assay is designed to be inclusive of all know allelic variants of stx and eae, including stx2g and stx2f. All reactions include an internal positive control (IPC) and ROX™ passive reference. Samples assessed as negative require no further testing. Samples assessed as positive must be further tested using the RapidFinder™ STEC Confirmation Assay.

The RapidFinder STEC Detection Workflow provides:
• Greater confidence in results—part of a validated workflow for detection of as little as 1-5 CFU of pathogenic STEC in your sample
• Simplified workflow—simultaneous detection of E. coli O157:H7 and the Big 6 non-O157:H7 strains in 375 g beef samples with a closed-tube, single-step lyophilized detection format that fits within your existing testing program
• Reduced chance of false results—through use of high-quality DNA preparation, proprietary assay design, and internal positive control
• Same day detection of pathogenic STECs and clearance of STEC-negative beef samples in as little as 12 hours, supporting early product release
• Minimized costs and laboratory space with the use a lower volume of a nonproprietary single-step enrichment medium, eliminating the need to purchase proprietary media
• Compatibility with RapidFinder™ Express software v.1.2 and later—enables easy, automated presence/absence detection and interpretation of results

Studies have shown that the RapidFinder STEC assays have a high level of sensitivity and specificity for E. coli O157:H7 and the Big 6 non-O157 STEC strains*. They detected all inclusion strains and showed no cross-reactivity to any of the exclusion strains tested. The stx assays detected all known variants of stx1 and stx2, including stx2f and stx2g. The optimized workflow co-developed with USDA-ARS showed equivalent detection to the MLG reference method, and the internal positive control in both assays significantly reduces the risk of false-negatives.

Applications
The RapidFinder STEC Detection Workflow is intended for use by microbiological analysts who need to test for STEC in beef samples. Combined with the Applied Biosystems 7500 Fast RT-PCR System, RapidFinder STEC screening and confirmation assays provide a comprehensive solution for rapid detection, identification, and confirmation of Big 6 STECs and E. coli 0157:H7 to meet importation and exportation requirements for the beef industry. Specific workflows are detailed in the figures below and include both automated and spin column-based DNA isolation and real-time PCR detection of E. coli O157:H7 and Big 6 non-O157 strains in beef samples.

RapidFinder Express Software
When integrated into the RapidFinder STEC Detection Workflow, RapidFinder Express Software automates data interpretation and generates user instructions on how to proceed with a sample. Data displays are available for novice and advanced users, showing simple presence/absence calls or raw data files using 7500 Fast System SDS Software.

RapidFinder STEC Screening and Confirmation Assays meet USDA FSIS guidance
The RapidFinder STEC Detection Workflow, a two-stage real-time PCR method, is designed to accurately detect, identify, and confirm E. coli O157:H7 and the Big 6 serogroups of non-O157 Shiga toxin-producing E. coli defined by the USDA Food Safety and Inspection Service (FSIS) as adulterants in the American beef industry.

Learn more about the RapidFinder STEC Detection Workflow >

*Swimley, Michelle S. Design and Evaluation of Two-Stage Multiplex Real-Time PCR Method for Detecting O157:H7 and non-O157 STEC Strains from Beef Samples. IAFP 2015.

RapidFinder™ Equine ID Kit

The RapidFinder™ Equine ID Kit enables the simple and reliable detection of horse meat in food and feed samples. The method works by detection of horse (Equus caballus) DNA by real-time PCR. Unlike ELISA based methods, the RapidFinder™ Equine ID Kit detects both raw and processed horse meat and provides sensitivity down to 0.01% horse meat DNA when used with the GMO Extraction Kit. A positive control is included in the kit (0.1% equine DNA) that can be used as a reference to set the threshold of detection of the assay.

• Simple—real-time PCR based assay, no electrophoresis needed
• Flexible—equine DNA detected in any food or feed sample, both raw and processed
• Reliable—internal positive control (IPC) provides process control check for PCR inhibitors
• Fast—from sample to result on the same day

How the RapidFinder™ Equine Kit Works
Equine DNA detection is accomplished by real-time PCR using the supplied primer/probe set specific for a mitochondrial DNA sequence that is unique to Equus caballus. The target sequence, detected in the FAM™ channel, is not only specific for horse, but also stable. Included in each reaction mix is an internal positive control detected in the VIC® channel, used to verify the absence of PCR inhibitors.

By performing a separate reaction with the included positive control, users are able to verify the system and assay are working correctly. More importantly, it enables easy data interpretation. The positive control is provided as 0.1% horse DNA in a background of beef DNA, and by comparing sample results to control results, a threshold of detection is provided.

The assay is compatible with Applied Biosystems® StepOne™, StepOnePlus™, 7500, 7500 Fast, and 7300 real-time PCR systems.

Sample Preparation
DNA sample preparation for the RapidFinder™ Equine ID Kit workflow is accomplished with the easy-to-use GMO Extraction Kit. Using 10 g of sample rather than the smaller sample sizes accommodated by most other DNA isolation procedures minimizes sampling errors. The GMO Extraction Kit is compatible with raw and processed meat, as well as composite food samples.

Quantification
If a positive result is obtained with the RapidFinder™ Equine ID Kit, you can quantify the amount of equine DNA in the sample using the RapidFinder™ Quant Equine Set.

Qubit™ RNA IQ Assay Kit (Invitrogen™)

The Qubit RNA IQ Assay Kit provides a fast, simple method to check whether an RNA sample has degraded using the Qubit 4 Fluorometer (required). The assay utilizes two unique dyes: one that binds to large, intact and/or highly structured RNA (mRNA, tRNA, rRNA), and another that selectively binds to small and/or, degraded RNA. Together they enable you to quickly assess the quality and integrity of an RNA sample.

To use, simply add your samples to the RNA IQ working solution, then measure on the Qubit 4 Fluorometer. Results are presented as an RNA IQ number (RNA IQ#) that indicates the RNA sample integrity and quality, and also as a calculated percent of large and small RNA in the sample. The RNA IQ# is a value from 1 to 10, similar to other RNA quality scores, where a small number indicates that the sample is comprised of mainly small RNA and a larger number indicates that the sample consists of mainly large RNA or RNA with tertiary structure.

Solution-based compared to electrophoresis methods
The RNA IQ assay results correlate well with electrophoretic methods. The primary difference between the solution-based RNA IQ assay and traditional electrophoresis-based techniques is the amount of time it takes to ascertain whether an RNA sample has degraded. Gels are considered easy to perform, but still take time to run. Microfluidic-based methods not only take time to obtain results, but often have lengthy procedures to prepare the samples. Gel or microfluidic-based electrophoresis is still recommended to obtain fragment size information or size distribution of the RNA sample.

Notes:
1. The Qubit RNA IQ assay will only work on the Qubit 4 Fluorometer. It will not work on the Qubit, Qubit 2.0, or Qubit 3 fluorometers.
2. 500 µL thin-walled PCR tubes (Cat. No. Q32856) are required but not included.

RapidFinder™ Poultry ID Kit

The RapidFinder™ Poultry ID Kit enables the simple and reliable detection of poultry meat in food and feed samples. The method works by detection of avian DNA, including DNA from chicken, turkey, duck, ostrich, goose, and other birds, by real-time PCR. Unlike ELISA based methods, when used with the GMO Extraction Kit (Cat. No. 4466336) for sample preparation, the RapidFinder™ Poultry ID Kit detects both raw and processed poultry meat and provides sensitivity down to 0.01% poultry DNA. The included positive control in the kit, which comprises 0.1% poultry DNA, can be used as a reference to set the threshold of detection of the assay.

• Simple-real-time PCR based assay, no electrophoresis needed
• Flexible-poultry DNA detected in any food or feed sample, both raw and processed
• Reliable-Internal Positive Control (IPC) provides process control check for PCR inhibitors
• Fast-from sample to result in the same day

How the RapidFinder™ Poultry ID Kit Works
Poultry DNA detection is accomplished by real-time PCR using the supplied primer/probe set targeting mitochondrial DNA sequence specific to several avian species, including but not limited to Gallus gallus (chicken), Meleagris gallopavo (turkey), Anas platyrhynchos (duck), Struthio camelus (ostrich), and Anser (goose). The primer/probes are supplied in a single tube, so if any of the poultry species is detected, a positive result will be obtained. Also included in each reaction mix is an internal positive control detected in the VIC® dye channel, which is used to verify the absence of PCR inhibitors.

By performing a separate reaction with the included positive control, users are able to verify the system and assay are working correctly. More importantly, it enables easy data interpretation. The positive control is provided as 0.1% Gallus gallus DNA in a background of fish DNA, and by comparing sample results to the control results, a threshold of detection is provided.

The assay is compatible with Applied Biosystems® StepOne™, StepOnePlus™, 7500, 7500 Fast, and 7300 real-time PCR systems.

Sample Preparation
DNA sample preparation for the RapidFinder™ Poultry ID Kit workflow is accomplished with the easy-to-use GMO Extraction Kit (Cat. No. 4466336). Using 10 g of sample rather than the smaller sample sizes accommodated by most other DNA isolation procedures minimizes sampling errors. The GMO Extraction Kit is compatible with raw and processed meat, as well as composite food samples.

Quantification
If a positive result is obtained with the RapidFinder™ Poultry ID Kit, most labs will go on to quantify the amount of poultry DNA in the sample. This can be accomplished using the RapidFinder™ Poultry ID Kit in conjunction with the RapidFinder™ Quant Multi-Meat Set (Cat. No. A24399).

Part of a Suite of Solutions For Meat Species Detection
The RapidFinder™ Poultry ID Kit is just one of our many available kits for the sensitive detection of various meat species in food and feed samples. Other RapidFinder™ meat species identification kits available include those for the detection of pork, beef, horse, sheep, ruminant, chicken, and turkey.

The RapidFinder™ Meat Species Identification Kits are made available through a partnership with IMEGEN (Instituto de Medicina Genomica).

ViewRNA™ Cell Plus Assay Kit (Invitrogen™)

The ViewRNA™ Cell Plus Assay is a novel assay that combines immunocytochemistry with ViewRNA™ technology, a proprietary fluorescent in situ hybridization and sequential branched-DNA amplification technique, to visualize both RNA with single-molecule sensitivity and protein in individual cells. This assay enables simultaneous detection of up to three RNA targets in combination with immunophenotyping for cell surface or intracellular proteins using both indirect and direct immunocytochemistry to allow for detailed characterization of specific cell subpopulations.

This ViewRNA™ Cell Plus Assay kit contains all the reagents needed to conduct the assay. Target Probe sets for genes of interest and antibodies are sold separately.

Reactivity/Species
Armenian Hamster, Baboon, Bovine, Canine, cat, Chicken, Chimpanzee, Cow, Cynomolgus Monkey, Donkey, Goat, Golden Syrian Hamster, Guinea Pig, Hamster, Horse, Human, Macaque, Monkey, Mouse, Non-human primate, Olive Baboon, Pig, Pigtailed Macaque, Rabbit, Rat, Rhesus, Sheep

Reported Application
Microscopy, Immunocytochemistry