Objective: To establish and evaluate microfluidic chip for rapid diagnosis of neurosurgical infections; Methods: Retrospective analysis of the pathogenic bacteria in Beijing Tiantan Hospital from 2006 to 2016 and CHINET data of bacterial neurosurgical infections, selecting pathogens and drug resistance genotypes as the parameters. The performance of the microfluidic chip was validated by 108 cerebrospinal fluid positive cultures. RESULTS: A total of 10 microbial identification and 15 resistance genes were included as the microfluidic chip parameters. The consistency assessment showed that the identification accuracy rate was amounted to 94.44%. The consistency of gene and phenotype with carbapenem antibiotics, clavulanic acid synergy test, oxacillin and vancomycin are higher, but the consistency of aminoglycoside antibiotics is low. Conclusion: The microfluidic chip is currently a very promising application technology for diagnosis of neurosurgical infections. It has rapid and accurate application properties and can be used as an important supplement to the clinical diagnosis.