Objective To investigate the distribution of virulence genes of Klebsiella pneumoniae(KPN)and to evaluate the diagnostic efficacy of peg-344 for hypervirulent strains.Methods KPN isolates were col-lected from January to December in 2023 in Xuanwu Hospital,Capital Medical University.Whonet5.6 was used to analyze the distribution and drug resistance of bacterial strains.Toxicity testing included wire string test,PCR amplification of virulence genes(peg-344,rmpA,iutA,iroB),K1 and K2 capsule serotypes,serum resistance test,and diagnostic efficacy of detection was analyzed by using receiver operating characteristic curve.Results A total of 122 KPN isolates were collected,including 45(36.9%)sensitive strains,20(16.4%)extended-spectrum β-lactamase positive strains,and 57(46.7%)carbapenem-resistant Klebsiella pneumoniae strains.The specimens were mainly isolated from sputum,urine,and blood,and mainly came from Department of Urology,Department of Neurology,Intensive Care Unit,and Department of Neurosurgery.The positive rate of string test was 27.9%.PCR detected K1(12.3%),K2(8.2%),peg-344(62.3%),rmpA(60.7%),and iutA(73.8%),but the iroB result was negative,and strains with multiple virulence genes accounted for 63.1%.The results of the serum resistance test showed that 43.5%,31.1%,and 25.4%of the samples were at levels 1-2,3-4,and 5-6,respectively.peg-344 had the largest area under the curve for diagnosing the hy-pervirulent strains,followed by peg-344+iutA and peg-344+rmpA.Conclusion KPN is widely distributed in the hospital,and there are a variety of virulence genes.peg-344 has clinical value in distinguishing low viru-lence from high virulence in KPN.