AccuSEQ Real-time PCR Software

The AccuSEQ software was developed to help optimize data interpretation when using our Applied Biosystems analytical solutions, as a component of an integrated workflow for detection of mycoplasma and adventitious viruses, and quantitation of residual DNA.

The AccuSEQ software was designed to facilitate the data interpretation of essential tests required for GMP manufacturing, including in-process testing and lot-release testing.

• Integrated, easy-to-use interface allows for consistency across multiple data applications and reduces the time needed to train your team
• Single software platform for all current and future resDNASEQ, MycoSEQ, and ViralSEQ real-time PCR assays
• Security, audit, and e-signature capabilities to help enable 21 CFR Part 11 compliance
• Available for 7500 Fast and QuantStudio 5, 0.1mL Block PCR instruments for easy integration of SEQ solutions into your current QC laboratory
   

The AccuSEQ Software v3.1 plate set up interface provides customers a visual approach to setting up their experiment details on a 96-well plate. Users can input details (sample, controls, dilutions, etc.) and designate colors for each well, allowing for quick visualization of the 96-well layout. Table-view plate setup is also available. The “Define & Setup Standard” feature allows users to expedite their standards set up on the plate. Simply enter information about the target and standard curve parameters, and this feature provides well arrangement on the plate for the Standards.

For rapid and accurate mycoplasma detection, the AccuSEQ software offers a plug-in analysis module for automated presence or absence calls when using the MycoSEQ Mycoplasma Detection system.

Key features and benefits:

• Optimized and validated for the MycoSEQ Mycoplasma Detection assay
• Automated detection of PCR inhibition in sample wells
• Automated detection of discriminatory positive control (DPC)
• Robust data analysis parameters for the MycoSEQ Mycoplasma Detection assay
• Flexibility for experiments; call settings may also be modified to meet the specifications of individual laboratories

The AccuSEQ Software v3.1 shows the “automated presence or absence calls” of mycoplasma contamination.

The AccuSEQ Software v3.1 shows the MycoSEQ “results analysis table” where calls are made based on the three acceptance criteria–C_t, T_m, and derivative value–of test samples and inhibition controls.

The AccuSEQ software offers factory default templates to facilitate experiment set up for the accurate and sensitive quantitation of host cell DNA impurities in both the purification process and in drug substance samples.

Key features and benefits:

• Streamlined resDNASEQ experiment setup (using factory default templates) and data reporting
• Automatic calculations of the resDNASEQ standard curve with curve fitting parameters
• Accurate quantitation of samples with % CV
• Computation of key metrics including % Recovery, Total DNA in pg/mL, and pg DNA/mg Protein
• Flexibility for custom experiments to meet the specifications of individual laboratories
• Customizable output layout that can be exported to a variety of formats
• Comprehensive reports which help eliminate the need for Excel sheets for calculations and preserve the audit trail

For plate setup using the AccuSEQ Software v3.1, the user simply enters sample names, serial dilution factor, and spike-in information.

After the experiment run, the AccuSEQ Software v3.1 automatically sets up the standard curve calculations and provides a visual display of the results with relevant curve fitting parameters, such as slope, intercept and efficiency, and R².

The AccuSEQ Software v3.1 shows the standard curve plot with samples overlayed and the results analysis tab on the right.

The resDNASEQ Results Table in AccuSEQ Software v3.1 shows key metrics including Quantity Mean, Quantity % CV, % Recovery, Total DNA per mL, and Total DNA per Protein Concentration.

The report section allows results to be exported in multiple formats (.xls, .xlsx, .txt, and .csv) and provides the ability to customize how this content appears in a printed report.

The AccuSEQ Software v3.1 showing the Print report section where users can customize report contents.
 

Snapshot of the resDNASEQ “Result Summary” from the PDF report generated by the v3.1 software. The Comprehensive Result Summary includes all important metrics required in the resDNASEQ experiment such as % CV, % Recovery, and Total DNA per mL (dilution adjusted).

For rapid and accurate MMV and vesivirus detection, the AccuSEQ software offers:

• Robust, optimized, and validated system—data analysis designed for use with ViralSEQ Detection Systems
• Automated detection—rapid assessment of PCR inhibition in sample wells
• Analysis options—identify true positive from false positives with discriminatory positive control
• Flexible call settings—meets specifications for unique lab requirements
   

The AccuSEQ software offers factory streamlined experimental set up for the accurate and sensitive quantitation of Sf-rhabodvirus contamination in both the purification process and in drug substance samples.

Key features and benefits:

• Streamlined experiment set-up—use factory default templates for easy data reporting
• Automated calculations—help eliminate the need for Excel sheets for calculations and preserve the audit trail
• Comprehensive reports—compute key metrics including percent recovery, total RNA in pg/mL, and pg RNA/mg Protein
• Flexibility—easily adjust for custom experiments to meet the specifications and reporting needs
   

There is also the custom experiment mode (“Create New”) where users can use the AccuSEQ software to create custom experiment setups. The custom mode guides the user through the experiment properties, qPCR run method details, and plate setup where there are easy-to-use tools for defining and setting up standards, controls, and unknowns. This custom workflow also supports ViralSEQ Mouse Minute Virus (MMV), vesivirus, and resDNASEQ SF9-Baculovirus duplex assays.

The AccuSEQ Software v3.1 shows the custom (“Create New”) selection where users have the ability to create an experiment with their own specific and unique parameters. 

The AccuSEQ Software v3.1 shows the custom (“Create New”) selection where users have the ability to create an experiment differing from the default experiment templates. Users can choose the experiment type, reagent type, and ramp speed.

Example of AccuSEQ 3.1 module for ViralSEQ MMV assay demonstrating the amplification readout.

Security, audit, and E-signature (SAE) is a key feature of the AccuSEQ software as it supports our customers in meeting their local regulations like 21 CFR Part 11 compliance. The software security, audit, and electronic signature (SAE) manager includes options that let you easily organize and track your security settings, such as:

• Security—username and password restrictions, and security policies such as password expiration and user account suspension
• Audit—automatically audits changes to experiments in the AccuSEQ Software and modifications to settings in the SAE module, and sorts and prints audit records at the experiment and systems level
• Electronic signature—selection of an electronic signature mode (enable or disable), on-demand electronic signature, electronic signature settings, number of electronic signatures required, and electronic signature actions (review, approve, or sign off)

Other key benefits include:

• Database storage for improved data security
• Customizable user role and permission
• Two levels of auditing including comprehensive reporting of audit events

Since AccuSEQ software is validated with respective real-time PCR systems (7500Fast and QuantStudio 5, 0.1mL block), Thermo Fisher Scientific offers compliance services like IQOQ (installation and operation qualification) and CSV (computer system validation) services that save customers time and money with compliance audits. For more information on compliance services, please contact your local sales or field support representative.

The AccuSEQ Software v3.1 shows a summary of the audit records.

The AccuSEQ Software v3.1 SAE Admin Console displays where user roles are defined and assigned.

This is an example of an audit report generated by the AccuSEQ Software v3.1.