Development of a Triple PCR Detection Method and Antibiotic Resistance Analysis for Micrococcus luteus Isolated from Fish

To identify strain fsznc-CL isolated from Pelteobagrus fulvidraco,establish a rapid detection method and discuss the drug sensitivity. The strain was identified by the morphological,physical and chemical characteristics and 16S rDNA sequence. Through the optimization of reaction system and conditions for three pairs of specific primers,a triple PCR detection method was established. This method was applied to the detection of thesamples infected Pelteobagrus fulvidraco. The drug resistance genes were amplified by PCR and the drug sensitivity of was detected by disk agar diffusion method. The results showed that strain fsznc-CL was Micrococcus luteus. Strain fsznc-CL was lethal to Pelteobagrus fulvidraco. Three pairs of target genes of Micrococcus luteus CBS,Sig and Pol could be accurately amplified by triple-PCR detection,while the other strains could not be amplified. The minimum template concentration for the detection of Micrococcus luteus was 5.822×10^-3 ng/μL. The positive rate of this method in detected sampleswas about 83.33%.This strain has drug resistance genes of TEM,aph(3')-Ⅱa,aac(6')-I,Sul1 and Sul2.Strain fsznc-CL was resistant to norfloxacin,oxacillin,erythromycin and furazolidone. This triple PCR method had high specificity and sensitivity. Micrococcus luteus was resistant to a few antibiotics at present.