Effect Of Compound Coptis Injection To MSW And Resistance Gene In Staphylococcus Aureus

The Staphylococcus aureus especially resistant Staphylococcus aureus have caused serious drug resistance problems to human and animal, which has become the most difficult one in the clinical treatment.In this thesis which investigated compound coptis injection to the Selection Window Mutation of sensitive Staphylococcus aureus and the grlA、gyrA gene sequences and NorA gene expression of resistant Staphylococcus aureus, it provided some basis study for further for injection as a resistant inhibitor of traditional Chinese medicine.The first part of the thesis we determined the MIC、MIC99and MPC in sensitive Staphylococcus aureus of Amoxicilli、Ceftiofur and Compound Coptis Injection single-used and combination used by typical MH agar double dilution.Conclusion showed that the MSW grew downwards obviously when (3-lactam-type drugs and Compound Coptis Injection combine used. The SI value is15.2of Amoxicillin alone,but combined with16MIC Compound Coptis Injection, the SI ruduce to5.1,became to its1/3of drug alone use; The SI value is16.1of Ceftiofur alone use.but combined with Compound Coptis Injection, the SI ruduce to4.2, became to its1/4of drug alone use.The second part of the thesis established the resistant-enrofloxcin、resistant-ofloxcin and resistant-norfloxcin Staphylococcus aureus by artificially induced.They had been detected its resistance by double dilution and disk diffusion methods;Then conducting the gene PCR amplification of grlA and gyrA which come DNA extraction reagent kit in sensitive Staphylococcus aureus ATCC29213、resistant-enrofloxcin、resistant-ofloxcin and resistant-norfloxcin Staphylococcus aureus by artificially induced and the processing lines which resistant strains had been dealed with Compound Coptis Injection, the amplification products connect with pMD18-T vector, transformed into JM109Escherichia coli competent cells, positive clones were screened, sequencing. The sequencing analysis showed that the gene base and amino acid sequence of grlA and gyrA all occurred mutation in resistant-enrofloxcin、resistant-ofloxcin and resistant-norfloxcin Staphylococcus aureus by artificially induced:After dealing with Compound Coptis Injection, the gene base and amino acid sequence of grlA and gyrA had producted obvious revertant,however it had not producted revertant for which resistant-norfloxcin strains had been dealed with Compound Coptis Injection.The third part of the thesis, it extract RNA by TRIZOL reagent in sensitive Staphylococcus aureus、resistant-enrofloxcin、resistant-ofloxcin、resistant-norfloxcin Staphylococcus aureus and the processing lines which resistant strains had been dealed with Compound Coptis Injection. Fluorescentquantitative RT-PCR had been used to study the impact of Compound Coptis Injection to NorA gene expression. The results show that the Compound Coptis Injection can reduce the expression of NorA gene mRNA of the resistant-enrofloxcin、resistant-ofloxcin and resistant-norfloxcin Staphylococcus aureus by artificially induced.