OBJECTIVE To establish a method of polymerase chain reaction(PCR) for detecting staphylococci in positive blood culture bottles.METHODS Genomic DNA in 493 positive blood culture bottles was extracted by guanidine hydrochloride and benzenemethanol,then genes 16S rRNA,ssa and mecA were amplified by PCR to identify staphylococci.Finally,the results of PCR were compared with that of traditional method.RESULTS To compare with traditional method,as the golden standard the sensitivity and specificity of PCR method were 98.6% and 100.0%,respectively,the detection could be finished in four hours.Method of PCR was better than traditional method in detecting meticillin-resistant staphylococci.CONCLUSIONS The PCR-based assay is simple,rapid,sensitive and specific,it can be used to detect staphylococci in positive blood culture bottles.

OBJECTIVE To establish a method of loop-mediated isothermal amplification(LAMP) for detecting Staphylococcus aureus in positive blood culture bottles.METHODS Genomic DNA in 293 positive blood culture bottles was extracted by guanidine hydrochloride and benzenemethanol,then genes ssa and mecA were amplified by LAMP to identify S.aureus.Finally,the results of LAMP were compared with the results of traditional method.RESULTS Twenty-two strains of S.aureus were detected in 293 positive blood culture bottles by LAMP method.Compared with traditional method,the sensitivity and specificity of LAMP method were both 100%,respectively,the detection could be finished in an hour.CONCLUSIONS The LAMP-based assay is simple,rapid,sensitive and specific which can be used to detect S.aureus in positive blood culture bottles rapidly.