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Adventitious Virus Testing
Regulatory agencies expect all biologics to undergo viral testing to reduce the risk of viral contamination. Sf-rhabdovirus, Mouse Minute Virus (MMV) and Vesivirus are all viral contaminants that are potential threats to drug development manufacturing processes. Get the reproducible accuracy and rapid results for risk mitigation or accelerated lot disposition with the ViralSEQ Real-Time PCR System that is part of a total workflow solution designed to lessen your bioproduction testing challenges in a GMP environment.
Sf-Rhabdovirus testing in gene therapy and vaccine production
It is important to have an accurate method for quantitating Sf-rhabdovirus which is a known infectious contaminant commonly found in Sf9/Sf21 cells used in gene therapy and vaccine development. The Applied Biosystems ViralSEQ Quantitative Sf-Rhabdovirus Kit accurately quantitates genomic Sf-rhabdovirus RNA.
This kit:
- Provides a negative-sense-specific RT-qPCR assay for accurate quantitation of genomic Sf-rhabodvirus RNA
- Has no cross-reactivity to positive-strand mRNA or other non-target species
- Is provided with RNA controls, 2 primer sets, for quantitation accuracy
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Step-by-step guide to the Sf-rhabdovirus quantitation workflow
Extract nucleic acid from sample matrices throughout the bioprocessing workflow using the PrepSEQ Nucleic Acid Sample Prep kit.
Add the purified RNA to the ViralSEQ Sf-Rhabdovirus kit assay mix to prepare the sample for reverse transcription
Perform reverse transcription with nucleic acid extracted and prepared in Step 1 to produce cDNA for further amplification for subsequent accurate quantitation of negative strain-specific DNA.
Run the 96-well plate containing the cDNA and reaction mix on a Real-Time PCR instrument.
Analyze the data using AccuSEQ Real-Time PCR Software, an intuitive easy-to-use analysis tool for data interpretation. The software guides the user through the experiment and plate setup and provides easy-to-use tools for defining and setting up standards, controls, and unknowns. AccuSEQ software is designed for analysis of results obtained from ViralSEQ, MycoSEQ, and resDNASEQ, kits.
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Extract nucleic acid from sample matrices throughout the bioprocessing workflow using the PrepSEQ Nucleic Acid Sample Prep kit.
Add the purified RNA to the ViralSEQ Sf-Rhabdovirus kit assay mix to prepare the sample for reverse transcription
Perform reverse transcription with nucleic acid extracted and prepared in Step 1 to produce cDNA for further amplification for subsequent accurate quantitation of negative strain-specific DNA.
Run the 96-well plate containing the cDNA and reaction mix on a Real-Time PCR instrument.
Analyze the data using AccuSEQ Real-Time PCR Software, an intuitive easy-to-use analysis tool for data interpretation. The software guides the user through the experiment and plate setup and provides easy-to-use tools for defining and setting up standards, controls, and unknowns. AccuSEQ software is designed for analysis of results obtained from ViralSEQ, MycoSEQ, and resDNASEQ, kits.
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Mouse Minute Virus (MMV) and Vesivirus testing for mammalian cell culture manufacturing processes
Multiple cell culture manufacturing facilities have been negatively impacted over the past 20 years following contamination with these viruses. The ViralSEQ Detection System is an integrated DNA purification and real-time PCR–based test for rapid, sensitive detection of MMV and Vesivirus from a wide range of sample types. The system detects all known MMV and Vesivirus strains and is compatible with the Prep SEQ 1-2-3 Nucleic Acid Extraction Kit, optimized for highly efficient DNA recovery.
- Rapid time-to-results, typically in less than 4 hours
- Specific with undetectable to no cross-reactivity with unrelated DNA
- Highly sensitive detection with TaqMan® technology for < 10 DNA copies per reaction (viral particles equivalent to 0.002 TCID50)
- Discriminatory positive/extraction DNA control minimizes false positive test results from cross-contamination with control DNA
High Sensitivity and Broad Dymanic Range.
The ViralSEQ MMV Assay can detect from 107 to 10 copies of MMV genomic DNA per reaction.
Step-by-step guide to the MMV and Vesivirus detection workflow
Extract nucleic acid (DNA for MMV or RNA for Vesivirus) using the PrepSEQ 1-2-3 Nucleic Acid Extraction kit. The DNA (for MMV detection) you purify in this step will be used as your template for Real-Time PCR,while the RNA (for Vesivirus detection) will be used as a template in a one-step Reverse Transcription Real-Time PCR.
Prepare the Real-Time PCR reactions by combining the purified DNA (MMV) or RNA (Vesivirus) with the TaqMan assay mix and master mix contained in the ViralSEQ MMV Detection Kit or ViralSEQ Vesivirus detection kit.
Run the 96-well plate containing extracted nucleic acid and reaction mix on a Real-Time PCR instrument.
Analyze the data using AccuSEQ Real-Time PCR Software, an intuitive easy-to-use analysis tool for data interpretation. The software guides the user through the experiment and plate setup and provides easy-to-use tools for defining and setting up standards, controls, and unknowns. AccuSEQ software is designed for analysis of results obtained from ViralSEQ, MycoSEQ, and resDNASEQ kits.
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Extract nucleic acid (DNA for MMV or RNA for Vesivirus) using the PrepSEQ 1-2-3 Nucleic Acid Extraction kit. The DNA (for MMV detection) you purify in this step will be used as your template for Real-Time PCR,while the RNA (for Vesivirus detection) will be used as a template in a one-step Reverse Transcription Real-Time PCR.
Prepare the Real-Time PCR reactions by combining the purified DNA (MMV) or RNA (Vesivirus) with the TaqMan assay mix and master mix contained in the ViralSEQ MMV Detection Kit or ViralSEQ Vesivirus detection kit.
Run the 96-well plate containing extracted nucleic acid and reaction mix on a Real-Time PCR instrument.
Analyze the data using AccuSEQ Real-Time PCR Software, an intuitive easy-to-use analysis tool for data interpretation. The software guides the user through the experiment and plate setup and provides easy-to-use tools for defining and setting up standards, controls, and unknowns. AccuSEQ software is designed for analysis of results obtained from ViralSEQ, MycoSEQ, and resDNASEQ kits.
Products
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