Objective To explore a rapid bacterial identifying method based on the 16S rRNA gene sequence analysis technology to provide the scientific basis for the diagnosis and treatment of unknown pathogenic bacteria.Methods The pure colonies were iso-lated and cultured directly from a clinical patient′s sputum sample.The colony as a template for PCR amplification with universal primers to amplify 16S rRNA gene fragments of unknown bacteria.The product of PCR was sequenced directly,then the sequence result was compared by using the BLAST of NCBI and the pathogen was identified based on the sequence homology.Results 1 strain of unknown pathogen was identified as ochrobactrum by this test and confirmed by ABI bacterial rapid identification sys-tem.Conclusion This study simplifies the isolation and identification procedures of unknown pathogen from the clinical samples and establishes a simple method for the rapid identification of pathogens by using 16S rRNA gene amplification.