Abstract:[Background] As the issue of lung health is aggravating, the emergence of hypervirulent Klebsiella pneumoniae (hvKP) has attracted widespread attention. As a variant of K. pneumoniae, hvKP is characterized by easy transmission, high virulence, and high fatality rate, which make it prone to causing community-acquired infection in healthy individuals. [Objective] To develop a sensitive and convenient detection method for hvKP, thus preventing its large-scale spread. [Methods] We developed a convenient and highly sensitive detection method for hvKP by combining recombinase polymerase isothermal amplification with the CRISPR ultra-sensitive detection system. [Results] This method was capable of detecting hvKP DNA samples as low as 58.1 fg/µL within 40 min without the need for enrichment. Moreover, it demonstrated excellent specificity, showing no amplification of 13 non-specific bacterial strains including classic K. pneumoniae. Furthermore, in the detection of simulated blood samples, this method showcased the sensitivity reaching 5.5 CFU/mL, which was two orders of magnitude higher than that (5.5×102 CFU/mL) of the conventional qRCR method. [Conclusion] This novel detection method has not only high sensitivity and specificity but also simple operation. This study aims to provide an effective tool for large-scale screening of hvKP in community-level medical institutions and offer a new detection scheme for public health services. This new technology is expected to play an important role in controlling and preventing the spread of hvKP.
