ViralSEQ™ Mouse Minute Virus (MMV) Detection System

Catalog number: 4444415

Applied Biosystems™  Related applications: Contaminant & Impurity QC Testing | Diagnostic Testing | Mammalian Cell Culture

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ViralSEQ™ Mouse Minute Virus (MMV) Detection Assay provides integrated sample preparation and real-time PCR assay for the detection of Mouse Minute Virus (MMV) in cell culture samples.
  • Detection of all known Mouse Minute Virus (MMV) strains
  • Proven TaqMan® technology for highly sensitive detection down to 10 DNA copies per reaction (viral particles equivalent to 0.002 TCID50)
  • Specific, no cross-reactivity with unrelated DNA
  • Discriminatory positive/extraction DNA control to eliminate possibility of false positive test results from accidental cross-contamination with control DNA
  • Use with Prep SEQ™ 1-2-3 Nucleic Acid Extraction Kit, which is optimized for highly efficient DNA recovery
  • Rapid time-to-results, typically in less than 4 hours
  • Part of the Cell Culture Rapid Methods Program

The Threat of MMV Contamination
MMV contamination is a potential threat to CHO and NS0 cell culture manufacturing processes. Multiple cell culture manufacturing facilities have been negatively impacted over the past 20 years following contamination with this virus. Use of a rapid, real-time PCR-based detection method enables in-process monitoring throughout the cell culture manufacturing process and helps to enable the earliest possible detection of a contamination event.

The Applied Biosystems® ViralSEQ™ MMV Detection Assay is an integrated DNA purification and real-time PCR-based test for rapid, sensitive detection of MMV from a wide range of sample types. First, viral DNA is isolated from the sample using PrepSEQ™ magnetic beads. Highly sensitive and specific detection of the viral DNA is then performed using TaqMan® real-time PCR technology. The assay also includes a proprietary positive control molecule that eliminates the possibility of a false positive test result arising from inadvertent contamination of the test sample with positive control.

Detection of All Known MMV
MMV is a member of the Parvovirus family and is a nonenveloped virus approximately 20 nm in diameter. MMV contains a single-stranded DNA that is roughly 5 kb in size. There are four strains of MMV (i, p, m, and c) and seven genomic sequences reported in the NCBI database. The ViralSEQ™ MMV Detection Assay was designed to help enable detection of all known strains and genomes.

Rapid Time-to-Results Facilitates Routine In-Process Testing
The ViralSEQ™ MMV Detection Assay has an easy workflow that can deliver results in just under 4 hours (Figure 1). The assay can be used to perform routine in-process testing, enabling multiple MMV tests throughout the cell expansion process. The rapid assay time allows the earliest possible detection of MMV contamination and helps enable rapid assessment of the contamination scope (such as testing bioreactors, related batches or lots, raw materials, and the facility). The scope contamination event can be bracketed immediately, and the affected equipment areas and/or suspect raw materials can be quarantined. In addition, rapid real-time PCR testing can be used to confirm decontamination and to provide verification of viral DNA removal prior to restarting the cell culture processes. The versatile protocol also accommodates testing of nontypical samples, including surface swabs, sampling ports, and probes. The ViralSEQ™ MMV Detection Assay streamlines the surveillance process for the earliest possible detection of recontamination (Figure 2).

Discriminatory Positive Control
The ViralSEQ™ MMV Detection Assay also includes the MMV Discriminatory Positive/Extraction Control, a plasmid DNA containing the MMV DNA sequence that is recognized by the MMV detection assay (FAM™ probe). This control (Figure 3) was designed to behave like MMV DNA in both the sample preparation and detection portions of the assay. It has been engineered, however, to incorporate an additional probe (VIC® probe) binding sequence. The presence of both FAM™ and VIC® signal in a PCR reaction can be used to discriminate a positive test result between MMV (FAM™) and the control DNA (FAM™ and VIC®). This novel control enables risk-free DNA spike control testing, eliminating the possibility of a false positive test result due to accidental cross-contamination of a test sample with the positive control DNA.

Proven Sensitivity and Specificity
The assay is highly sensitive, able to detect as few as 10 copies of MMV genomic DNA per PCR reaction, which is equivalent to 0.002 TCID50. Analysis of 10-fold serial dilutions of MMV genomic DNA demonstrates the broad dynamic range and efficiency of the assay (Figures 4 and 5).

Table 1 shows the ViralSEQ ™ MMV Assay to be specific to the MMV genomic DNA target with no cross-reactivity with unrelated DNA.

Highly Efficient Recovery of MMV Genomic DNA Using the PrepSEQ™ 1-2-3 Sample Preparation Kit
The PrepSEQ™ 1-2-3 Nucleic Acid Extraction Kit is designed to help enable consistent and high percentage recovery of the MMV genomic DNA sample from complex sample types that include CHO cell culture and bovine serum. Figure 6 shows the Ct values of DNA extracted from the spiked sample were comparable to unspiked control that was amplified in the real-time PCR reaction, demonstrating the highly efficient DNA recovery using the PrepSEQ™ 1-2-3 Kit.

Cell Culture Rapid Methods Program
The ViralSEQ™ MMV Detection Assay is part of the Cell Culture Rapid Methods Program, designed to streamline the detection of three common contaminants of mammalian cell culture-based biopharmaceutical manufacturing. The program sets new workflow standards in efficiency and product quality, combining one sample preparation step with real-time PCR-based assays for the detection of Mycoplasma, Vesivirus, and MMV on one instrument platform.
For Research Use Only. Not for use in diagnostic procedures.


Detection Method: Primer-Probe Detection
PCR Method: qPCR
Product Line: Applied Biosystems™, ViralSEQ™
Product Size: 100 reactions
Sample Type (Specific): Viral DNA
Sensitivity: ≥ 10 copies MMV DNA
Species: Mouse
Target Organism Class: Mouse Minute Virus (MMV)
Target Specificity: Target-Specific
Shipping Condition: Dry Ice

Contents & storage

Contains Real-Time PCR assay with buffer and enzyme mix, positive control and negative controls


Manuals & protocols