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Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Updated: Jun 21, 2025

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Published on: March 30, 2015

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

A R MacLean¹, R Gunson¹

¹West of Scotland Specialist Virology Centre, New Lister Building, Glasgow Royal Infirmary, 10-16 Alexandra Parade, Glasgow G31 2ER, United Kingdom.

Journal of Virological Methods

|July 14, 2024

Summary

This summary is machine-generated.

PCR.Ai accurately automates multiplex quantitative real-time PCR (qPCR) analysis for pathogen testing. This tool significantly reduces hands-on time and complexity, enabling faster, more cost-effective, and reliable results in clinical diagnostics.

Keywords:

Multiplex PCR PCR:Ai Quantitation

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Last Updated: Jun 21, 2025

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

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Published on: March 30, 2015

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

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Published on: August 3, 2011

Author Spotlight: Advancements in Multiplex Detection of Respiratory Viruses

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Published on: November 10, 2023

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.