Valencia-Bacca Juan D, Islam Md Maidul, Haas Karen M, Zafar M Ammar
Department of Microbiology and Immunology, Wake Forest University School of Medicine, Winston-Salem, North Carolina.
These authors contributed equally to this work.
Curr Protoc. 2025 Aug;5(8):e70187. doi: 10.1002/cpz1.70187.
Klebsiella pneumoniae (Kpn) is a Gram-negative opportunistic pathogen and a leading cause of hospital-acquired infections. The gastrointestinal (GI) tract serves as a critical site for Kpn colonization and a reservoir for host-to-host spread that primarily occurs through the fecal-oral route. The emergence of multidrug-resistant strains, coupled with virulence factors such as enhanced capsule production, presents significant challenges for clinical management and public health. Animal models-particularly murine models of GI colonization-are essential for advancing our understanding of Kpn pathogenesis, host-pathogen interactions, and for identifying therapeutic strategies. In this manuscript, we describe standardized protocols for studying Kpn GI colonization and systemic dissemination. These methods are designed to facilitate reproducibility and consistency across laboratories investigating Kpn colonization dynamics, dissemination routes, and immune evasion mechanisms. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. By providing detailed and optimized experimental approaches, this work aims to support the development of uniform methodologies for Kpn GI colonization and host response studies Basic Protocol 1: Murine model of Kpn gastrointestinal colonization via oral feeding Basic Protocol 2: Murine model of Kpn gastrointestinal colonization via gavage Basic Protocol 3: Murine model of Kpn intraperitoneal challenge Basic Protocol 4: Murine model of systemic Kpn infection via intravenous injection Basic Protocol 5: Kpn transmission studies following gastrointestinal colonization with intact microbiota Basic Protocol 6: Transmission studies of Kpn gastrointestinal colonization in antibiotic-pretreated mice Support Protocol 1: Preparation of Kpn inoculum Support Protocol 2: Preparation of mixed Kpn inoculum for competitive infection Support Protocol 3: Assessment of Kpn bacterial burden Support Protocol 4: Preparation of cecal filtrate.
肺炎克雷伯菌（Kpn）是一种革兰氏阴性机会致病菌，是医院获得性感染的主要原因。胃肠道是Kpn定植的关键部位，也是主要通过粪-口途径进行宿主间传播的储存库。多重耐药菌株的出现，加上如增强荚膜产生等毒力因子，给临床管理和公共卫生带来了重大挑战。动物模型——尤其是胃肠道定植的小鼠模型——对于增进我们对Kpn发病机制、宿主-病原体相互作用的理解以及确定治疗策略至关重要。在本手稿中，我们描述了研究Kpn胃肠道定植和全身播散的标准化方案。这些方法旨在促进各实验室在研究Kpn定植动态、传播途径和免疫逃逸机制方面的可重复性和一致性。© 2025作者。由Wiley Periodicals LLC出版的《当前方案》。通过提供详细且优化的实验方法，这项工作旨在支持开发用于Kpn胃肠道定植和宿主反应研究的统一方法。基本方案1：通过口服喂养建立Kpn胃肠道定植的小鼠模型。基本方案2：通过灌胃建立Kpn胃肠道定植的小鼠模型。基本方案3：Kpn腹腔注射攻击的小鼠模型。基本方案4：通过静脉注射建立Kpn全身感染的小鼠模型。基本方案5：完整微生物群存在下胃肠道定植后的Kpn传播研究。基本方案6：抗生素预处理小鼠中Kpn胃肠道定植的传播研究。支持方案1：Kpn接种物的制备。支持方案2：用于竞争性感染的混合Kpn接种物的制备。支持方案3：Kpn细菌负荷的评估。支持方案4：盲肠滤液的制备。
