Milner Centre for Evolution, University of Bath, Bath, United Kingdom.
Department of Biology and Biochemistry, University of Bath, Bath, United Kingdom.
PLoS One. 2020 Apr 30;15(4):e0232334. doi: 10.1371/journal.pone.0232334. eCollection 2020.
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.
百日咳博德特氏菌是百日咳的病原体，通常称为百日咳。尽管通过疫苗接种降低了百日咳的发病率，但在过去三十年中，许多国家的百日咳发病率又回到了很高的水平。这种反弹与全细胞疫苗向无细胞疫苗的转变有关。与全细胞疫苗相比，无细胞疫苗提供的保护似乎是短暂的。为了通过确定保护的免疫相关性来为未来的疫苗改进提供信息，已经开发了一种百日咳博德特氏菌定植的人体挑战模型。为了在该模型中准确测量定植状态，需要开发一种基于 qPCR 的检测方法来对样本中的百日咳博德特氏菌进行计数，该方法能够区分活细菌和死细菌。在这里，我们报告了该检测方法的开发及其在人体挑战模型样本中定量检测百日咳博德特氏菌的性能。该检测方法在需要定量测量百日咳博德特氏菌数量和活力的诊断实验室和研究中具有未来的应用价值。
