Abstract

Postoperative intracranial infection represents a significant complication in neurosurgical procedures, with Acinetobacter baumannii (Ab) and Klebsiella pneumoniae (Kp) frequently identified as common drug-resistant pathogens. In response to this challenge, we have developed an efficient, extraction-free detection method that facilitates the rapid and highly sensitive identification of Ab and Kp. This method employs a single-tube format that integrates heat lysis with PCR, allowing for the concurrent bacterial lysis and target gene amplification. Detection of the amplified products is achieved through the Cas12a/crRNA complex, which, upon activation, exhibits trans-cleavage activity that cleaves reporters to produce a signal detectable by microplate readers or lateral flow test strips (LFTS). The Heat Lysis-PCR-CRISPR/Cas12a (HLPCR-CRISPR/Cas12a) detection platform unifies lysis and amplification in one step, streamlining the process. It exhibits high specificity (no cross-reactivity) and achieves detection limits of 10 2  CFU/μL (fluorescence) and 10 3  CFU/μL (LFTS). When validated on clinical CSF samples, it delivered results in 90 min with perfect concordance with quantitative PCR (qPCR). Combining speed, high sensitivity, specificity, and operational simplicity, this platform provides a powerful diagnostic tool for early detection of Ab and Kp in postoperative intracranial infections.