Abstract: OBJECTIVE To evaluate the clinical value of polymerase chain reaction (PCR)-microarray hybridization in the diagnosis of bloodstream infection. METHODS Total of 316 positive blood cultures were collected, including 161 gram-positive blood cultures and 155 gram-negative blood cultures. After the positive blood culture was detected, the culture medium was collected in time for routine culture identification, drug sensitivity and PCR-microarray hybridization. The specimens which showed the inconsistent results were analyzed by sequencing and the specimen turn-around time (TAT) was calculated. The results were statistically analyzed. RESULTS Nineteen gram-positive strains (161 strains) were identified by blood culture, among which 67 strains were resistant to oxacillin. There were 13 strains of gram-negative bacteria (155 strains), including 36 extended spectrum β lactamases (ESBLs)-producing strains and 14 carbapenem-resistant Klebsiella pneumoniae (KPC) strains. Five gram-positive strains (148 strains) were detected by PCR-microarray hybridization, including 67 strains of drug-resistant gene MecA. There were 6 strains of gram-negative bacteria (147 strains) , including 27 strains of CTX-M-1 and 14 strains of KPC. The sensitivity and specificity of PCR-microarray hybridization were 85.7%-100.0% and 99.1%-100.0%. respectively. The sensitivity and specificity of MecA and KPC were 100.0%, and those of CTX-M-1 were 69.4% and 97.0%. The average shortening time of TAT for gram-positive and gram-negative bacteria was 39.52 h and 38.08 h, respectively. CONCLUSION PCR-microarray hybridization shows high sensitivity and specificity for identification of strains and detection of drug-resistant genes in positive blood culture medium, which can effectively shorten TAT and has a good prospect of clinical application.