Main Drug Resistance Gene Clone And Research Of Homology Of Staphylococcus Aureus From Animals

In recent years, the resistance of antibiotic has received more and more concern for principal factorsthat includes the widespread, sometimes inappropriate and continual use of antimicrobials.Especially drugresistance of Staphylococcus aureus effect on therapeutic efficacy of anti-infection, which was one of thecommon pathogenic bacteria on clinic. Accordingly, it is imminent to detecting drug resistance and tocontrol dissemination.Gene chip was an efficient technology to determine the drug resistance of pathogen quickly. It couldsatisfy the requirement of detecting drug resistance with characteristics of high-flux, quick, high-flux andhigh-specificity. In my experiment, gathering S.aureus from animals,detecting drug resistance,cloning drugreistance genes and researching the homology prepare for positive sample copy of drug resistance genechip. The research is consisted of three contents:14 S.aureus were isolated and identificated from pus, blood of Septicemia, liver, kidney, and spleen ofmorbid domestic animals and domestic fowls in regions of Jilin province, they were researched forprepareing positive specimen of detected drug resistance gene chip for S.aureus.Detecting by plate double dilution for penicillinum G, Erythromycin, Lincomycin, Ciprofloxacin andAchromycin of Minimal Inhibitory Concentration (MIC). As a result, 1 strain was resistant forpenicillinum G, 7 isolates were resistant for Erythromycin, Lincomycin,4 isolates were resistant forCiprofloxacin and 6 strains were resistant for vibramycin.6 strains were multidrug resistant.We selected mecA gene and femA factor asβ-lactam antibiotic-resistant genes, ermA, ermC, msrAgene as macrolides-lincosamids antibiotic-resistant genes, norA, grlA, gyrA, gene as Quinolonesantibiotic-resistant genes, and tetM gene as Tetracycline antibiotic-resistant genes on the basis of the latestresearch. We designed primers with GenBank enunciable gene sequence, cloned the drug resistantgenes, and sequenced these genes. The homology of nucleotide sequence was analyzed with DNAsissoftware. The result showed that the homology was up to 99％compared with publicated sequence.MecAof S. aureus H34, ermA of S. aureus H34, ermC of S. aureus O41, msrAof S. aureus H34 and K37, gyrA, grlAof S. aureus H34, norA of S. aureus S19 and terM of S. aureus H34 consider to be the positive specimen. Wefind the 96 and 99 of grlA in QRDR of S. aureus H34 have mutated from Leu to Ser for the first time. Itmay have relatations with drug resistance for Quinolones in S.aureus.The two mutations will be consideredas the positive specimen of detecting drug resistance gene chip.