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Published on: February 1, 2014
A qPCR assay for Bordetella pertussis cells that enumerates both live and dead bacteria
Stacy Ramkissoon1,2, Iain MacArthur1,2, Muktar Ibrahim3,4,5,6
1Milner Centre for Evolution, University of Bath, Bath, United Kingdom.
Abstract:
Bordetella pertussis is the causative agent of whooping cough, commonly referred to as pertussis. Although the incidence of pertussis was reduced through vaccination, during the last thirty years it has returned to high levels in a number of countries. This resurgence has been linked to the switch from the use of whole-cell to acellular vaccines. Protection afforded by acellular vaccines appears to be short-lived compared to that afforded by whole cell vaccines. In order to inform future vaccine improvement by identifying immune correlates of protection, a human challenge model of B. pertussis colonisation has been developed. Accurate measurement of colonisation status in this model has required development of a qPCR-based assay to enumerate B. pertussis in samples that distinguishes between viable and dead bacteria. Here we report the development of this assay and its performance in the quantification of B. pertussis from human challenge model samples. This assay has future utility in diagnostic labs and in research where a quantitative measure of both B. pertussis number and viability is required.
Insights
A new qPCR assay accurately measures viable Bordetella pertussis (whooping cough bacteria) in human samples. This tool aids research into vaccine effectiveness and pertussis resurgence.
Area of Science:
- Microbiology
- Immunology
- Vaccinology
Background:
- Bordetella pertussis causes whooping cough (pertussis).
- Pertussis incidence has resurged globally, linked to acellular vaccines with shorter protection durations than whole-cell vaccines.
- A human challenge model is crucial for studying B. pertussis colonization and vaccine correlates of protection.
Purpose of the Study:
- To develop and validate a quantitative PCR (qPCR) assay for enumerating viable B. pertussis.
- To accurately measure B. pertussis colonization in a human challenge model.
- To support the identification of immune correlates for improved pertussis vaccines.
Main Methods:
- Development of a novel qPCR assay.
- Enumeration of B. pertussis in human samples from a challenge model.
- Distinguishing between viable and dead B. pertussis bacteria.
Main Results:
- The developed qPCR assay accurately quantifies B. pertussis in human challenge model samples.
- The assay can differentiate between live and dead bacteria, providing a measure of viability.
- Performance characteristics of the assay were established.
Conclusions:
- The validated qPCR assay is a valuable tool for assessing B. pertussis colonization.
- This assay will aid research into pertussis pathogenesis and vaccine efficacy.
- Future applications include diagnostic laboratories and research requiring quantitative viability data.
