One-Pot Isothermal LAMP-CRISPR-Based Assay for Klebsiella pneumoniae Detection

作者：Xiaotong Qiu, Xueping Liu, Xiao Ma, Ruixue Wang, Shenglin Chen, Fang Li, Zhenjun Li · 发表于：Microbiology Spectrum · 年份：2022 · DOI：10.1128/spectrum.01545-22 · 被引用次数：45 · 研究领域：CRISPR and Genetic Engineering、Biosensors and Analytical Detection、Advanced biosensing and bioanalysis techniques

Klebsiella pneumoniae is a significant threat to global health. At present, the methods of K. pneumoniae detection are culture-based and instrument-dependent and are not suitable for rapid diagnostic. This study reports K. pneumoniae CRISPR-top assay, which can precisely identify K. pneumoniae using nucleic acids of pure cultures or clinical samples in one pot with one fluid-handling step. The K. pneumoniae CRISPR-top reaction can be completed within 60 min at a constant temperature, thus specific instruments are not required. Our results show that CRISPR-top assay yields enormous advantages compared with LAMP detection. The K. pneumoniae CRISPR-top assay can be a high-efficiency alternative tool for rapid and accurate diagnosis of K. pneumoniae infection, especially in resource-limited settings.