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Towards improved accuracy of Bordetella pertussis nucleic acid amplification tests
1Department of Pathology, University of Texas Medical Branch, Galveston, Texas, USA. mjloeffe@utmb.edu
Abstract:
In many clinical microbiology laboratories, nucleic acid amplification tests such as PCR have become the routine methods for the diagnosis of pertussis. While PCR has greatly increased the ability of laboratories to detect Bordetella pertussis infections, it has also been associated with false-positive results that can, given the tendency of B. pertussis to cause outbreaks, result in unnecessary and costly control measures. The species specificity of Bordetella gene targets and their number of copies per genome greatly impact the performance characteristics of nucleic acid amplification tests for B. pertussis. It is crucial that laboratorians recognize these characteristics, to limit false-positive test results and prevent pseudo-outbreaks.
Insights
PCR tests for pertussis are common but can yield false positives. Understanding Bordetella gene targets is key to accurate Bordetella pertussis detection and preventing unnecessary outbreak responses.
Area of Science:
- Clinical Microbiology
- Molecular Diagnostics
- Infectious Disease Epidemiology
Background:
- Nucleic acid amplification tests, including PCR, are standard for diagnosing pertussis.
- PCR enhances detection of Bordetella pertussis but is prone to false-positive results.
- False positives can trigger costly and unnecessary control measures during pertussis outbreaks.
Purpose of the Study:
- To highlight the impact of Bordetella gene target selection on PCR accuracy for pertussis.
- To emphasize the importance of understanding species specificity and copy number for B. pertussis detection.
- To guide laboratorians in minimizing false-positive PCR results and preventing pseudo-outbreaks.
Main Methods:
- Analysis of species specificity of Bordetella gene targets used in PCR assays.
- Evaluation of the impact of target gene copy number per genome on assay sensitivity and specificity.
- Review of clinical laboratory data and outbreak investigation protocols related to pertussis diagnosis.
Main Results:
- The choice of Bordetella gene targets significantly influences the specificity of PCR assays.
- Higher copy numbers of target genes can increase sensitivity but may also elevate false-positive rates.
- Inappropriate target selection can lead to misidentification and pseudo-outbreaks.
Conclusions:
- Careful selection of species-specific Bordetella gene targets is critical for accurate pertussis diagnosis via PCR.
- Laboratorians must be aware of assay characteristics to prevent false-positive results.
- Accurate molecular diagnostics are essential for effective pertussis surveillance and outbreak management.
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