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Updated: Jun 4, 2026
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Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
Published on: March 30, 2015
Proof of concept for recombinant cellular controls in quantitative molecular pathogen detection
Peter Rossmanith1, Patrick Mester, Karin Frühwirth
1Christian Doppler Laboratory for Molecular Food Analytics, Veterinärplatz 1, 1210 Vienna, Austria. peter.rossmanith@vetmeduni.ac.at
Applied and Environmental Microbiology
|February 15, 2011
Summary
This study introduces internal sample process controls (ISPCs) to track analytical chain efficiency. A novel recombinant Listeria strain effectively demonstrated ISPC suitability for accurate molecular testing.
Area of Science:
- Food safety microbiology
- Molecular diagnostics
- Analytical chemistry
Background:
- Monitoring the efficiency of analytical methods is crucial for reliable results.
- Quantitative PCR (qPCR) is widely used but requires robust process controls.
- Traditional controls may not fully capture sample preparation variability.
Purpose of the Study:
- To introduce and validate internal sample process controls (ISPCs).
- To assess the analytical suitability of a novel recombinant strain as an ISPC.
- To monitor the entire analytical chain, including sample preparation and qPCR.
Main Methods:
- Development of a recombinant Listeria monocytogenes strain with a deleted prfA gene.
- Incorporation of a competitive artificial single-copy genomic target within the recombinant strain.
- Application of the ISPC in naturally contaminated food samples.
Main Results:
- The developed ISPC demonstrated analytical suitability for monitoring qPCR efficiency.
- The recombinant strain effectively tracked the entire analytical process.
- Successful application in real-world, naturally contaminated samples.
Conclusions:
- Internal sample process controls (ISPCs) offer a reliable method for monitoring analytical chain performance.
- The novel recombinant Listeria strain serves as a validated ISPC for molecular assays.
- This approach enhances the accuracy and trustworthiness of diagnostic testing.
