Related Experiment Video
Updated: Aug 18, 2026
Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
Published on: March 30, 2015
Development and use of an internal positive control for detection of Bordetella pertussis by PCR
Stéphanie Herwegh1, Christophe Carnoy, Frédéric Wallet
1Laboratoire de Bactériologie-Hygiène, Hôpital A. Calmette, F-59037-Lille Cedex, France.
Abstract:
An internal control of amplification was constructed by recombinant PCR to detect PCR inhibitors. This exogenous DNA was included in the reaction mixture and coamplified with the target gene. This detection was successfully applied to the diagnosis of whooping cough by amplification of a fragment of Bordetella pertussis IS481.
Insights
An internal control using recombinant PCR was developed to detect PCR inhibitors. This method was successfully applied to diagnose whooping cough by amplifying Bordetella pertussis DNA.
Area of Science:
- Molecular Biology
- Diagnostic Microbiology
Background:
- Polymerase Chain Reaction (PCR) is a powerful tool for DNA amplification.
- The presence of PCR inhibitors in clinical samples can lead to false-negative results.
- Accurate detection of infectious agents requires reliable amplification methods.
Purpose of the Study:
- To develop and validate an internal control for PCR amplification.
- To assess the utility of this control in detecting PCR inhibitors.
- To apply the method for the diagnosis of whooping cough.
Main Methods:
- Construction of an exogenous DNA internal control using recombinant PCR.
- Coamplification of the internal control with the target gene in the same reaction mixture.
- Application of the method to detect Bordetella pertussis IS481 DNA.
Main Results:
- The internal control effectively detected the presence of PCR inhibitors.
- The method demonstrated successful coamplification with the target gene.
- The assay was successfully applied to the diagnosis of whooping cough.
Conclusions:
- Recombinant PCR-based internal controls are effective for detecting PCR inhibitors.
- This approach enhances the reliability of PCR-based diagnostic tests.
- The developed method provides a robust tool for whooping cough diagnosis.
