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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: May 19, 2026

Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres

Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres

Published on: October 23, 2011

Quantitative Multiplex Digital PCR with Fluorescence-Encoded Nanoreactor Beads.

Stephan Hubold¹, Lea Kanitz¹, Oliver Lemuth¹

¹BLINK AG, Bruesseler Strasse 20, 07747 Jena, Germany.

Analytical Chemistry

|May 18, 2026

Summary

This summary is machine-generated.

We developed a novel multiplex digital PCR method using fluorescence-encoded magnetic Nanoreactor Beads (femNRB). This adaptable platform simplifies the creation and expansion of diagnostic test panels for precision medicine.

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Area of Science:

Molecular Biology
Biotechnology
Diagnostic Assays

Background:

Syndromic PCR panels are crucial for precision medicine but are difficult to design and validate.
Integrating new markers into existing panels, especially quantitative assays, requires extensive revalidation.

Purpose of the Study:

To introduce a novel multiplex digital PCR strategy for adaptable and scalable diagnostic panels.
To enable cross-reactivity-free detection and absolute quantification of multiple genetic markers simultaneously.

Main Methods:

Utilized fluorescence-encoded magnetic Nanoreactor Beads (femNRB) for multiplex digital PCR.
Developed a system with reversibly conjugated primers and probes for adaptable panel assembly.
Validated a 10-assay panel for nosocomial pathogens, later expanded to 16 assays with antibiotic resistance and virulence markers.

Main Results:

Demonstrated cross-reactivity-free detection and absolute quantification of multiple targets in a single sample.
Achieved equivalent analytical performance for both the 10- and 16-assay panels without further optimization.
Showcased the robustness, scalability, and modularity of the femNRB-based platform.

Conclusions:

The femNRB approach offers a powerful, modular solution for developing and upgrading diagnostic PCR panels with minimal effort.
This method simplifies assay expansion by eliminating the need for recalibration, supporting seamless integration of new markers.
The platform enables uncompromised quantification across variable sample concentrations without complex hardware.