Objective To establish the proteomic diagnostic model for Klebsiella pneumoniae strains producing extended spectrum beta-lactamases(ESBLs) and search for a rapid method to identify them. Methods Klebsiella pneumoniae proteins were detected by the PBS Ⅱ/C protein fingerprint spectrometer and Au protein chip.The differentially expressed proteins of Klebsiella pneumoniae ESBLs positive and negative strains were analyzed and an artificial neural network(ANN) model of Klebsiella pneumoniae ESBLs producing strains was established and validated. Results Totally,47 protein peaks were detected for the moleculars with mass-to-charge ratio(m/z) within the range of 2,000-20,000.The peaks with m/z of 5,985.5,8,361.6 and 9,169.8 were screened by ANN as the protein spectrum model of Klebsiella pneumoniae ESBLs producing strains.The validation result of the model indicated that the sensitivity was 96.88% and the specificity was 93.62%. Conclusions The protein spectrum model established by this study can identify Klebsiella pneumoniae ESBLs producing strains accurately and rapidly.It is worth further studying.