Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden.
Methods Mol Biol. 2023;2674:101-115. doi: 10.1007/978-1-0716-3243-7_7.
Pathogenic bacteria have evolved the ability to evade their host defenses and cause diseases. Virulence factors encompass a wide range of adaptations that allow pathogens to survive and proliferate in the hostile host environment during successful infection. In human pathogenic Yersinia species, the potent type III secretion system (T3SS) and other essential virulence factors are encoded on a virulence plasmid. Here, we investigated the bacterial growth rate and plasmid copy number following a Yersinia infection using droplet digital PCR (ddPCR). ddPCR is an exceptionally sensitive, highly precise, and cost-efficient method. It enables precise quantification even from very small amounts of target DNA. This method also enables analysis of complex samples with large amounts of interfering DNA, such as infected tissues or microbiome studies.
病原细菌已经进化出逃避宿主防御并导致疾病的能力。毒力因子包含了广泛的适应性，使病原体能够在成功感染期间在宿主环境中存活和繁殖。在人类病原性耶尔森氏菌中，强大的 III 型分泌系统（T3SS）和其他必需的毒力因子都编码在一个毒性质粒上。在这里，我们使用液滴数字 PCR（ddPCR）研究了耶尔森氏菌感染后细菌的生长速度和质粒拷贝数。ddPCR 是一种非常灵敏、高度精确和具有成本效益的方法。它即使从非常少量的目标 DNA 也能进行精确的定量。这种方法还可以分析含有大量干扰 DNA 的复杂样本，如感染组织或微生物组研究。
