Automation and standardisation of a quantitative multiplex PCR assay using PCR.Ai

Area of Science:

• Clinical diagnostics
• Molecular biology
• Bioinformatics

Background:

• Routine clinical pathogen testing involves manual data analysis and quality control for quantitative real-time PCR (qPCR).
• Previous studies demonstrated 100% concurrence between manual methods and PCR.Ai for non-quantitative qPCR.
• This study further evaluates PCR.Ai's accuracy and impact on quantitative multiplex qPCR.

Purpose of the Study:

• To evaluate the impact of PCR.Ai as a final interpretation/verification step for routine in-house multiplex quantitative qPCR tests.
• To assess PCR.Ai's accuracy and time-saving potential compared to manual interpretation.
• To analyze 1350 interpretations for Cytomegalovirus (CMV), Epstein-Barr virus (EBV), and adenovirus in blood samples.

Main Methods:

• Comparison of PCR.Ai with existing manual interpretation methods.
• Assessment of accuracy in detection and quantitation of CMV, EBV, and adenovirus.
• Measurement of hands-on time savings per run.

Main Results:

• 100% concurrence achieved between manual analysis and PCR.Ai for CMV, EBV, and adenovirus detection and quantitation.
• Significant time savings of 63 minutes per run demonstrated with PCR.Ai.
• PCR.Ai proved to be a highly accurate and time-saving tool.

Conclusions:

• PCR.Ai simplifies qPCR analysis, reducing the need for specialists and hands-on time.
• The tool enables quicker results, lower costs, and reduced risk of errors in quantitative PCR testing.
• PCR.Ai is a valuable asset for enhancing efficiency and accuracy in clinical diagnostic laboratories.