本文關(guān)鍵詞：肺炎克雷伯菌耐藥突變選擇窗的體內(nèi)外研究　出處：《中國(guó)醫(yī)科大學(xué)》2010年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 肺炎克雷伯菌 左氧氟沙星 防突變濃度 突變選擇窗

【摘要】： 目的 體外測(cè)定氟喹諾酮類(lèi)藥物左氧氟沙星對(duì)臨床分離肺炎克雷伯菌的最低抑菌濃度(Minimum Inhibitory Concentration, MIC)和防突變濃度(Mutant Prevention Concentration, MPO)。建立小鼠肺部肺炎克雷伯菌感染模型,測(cè)定相關(guān)藥代／藥效動(dòng)力學(xué)(PK/PD)參數(shù),觀察藥物濃度對(duì)耐藥突變菌株富集程度的影響,結(jié)合體外實(shí)驗(yàn)測(cè)定結(jié)果,在動(dòng)物體內(nèi)初步驗(yàn)證突變選擇窗(MSW)理論,并通過(guò)MSW理論與已有的藥代動(dòng)力學(xué)參數(shù),對(duì)臨床左氧氟沙星治療肺炎克雷伯菌肺炎的給藥策略進(jìn)行評(píng)價(jià)。 方法 采用CLSI規(guī)定的標(biāo)準(zhǔn)瓊脂平皿二倍稀釋法,測(cè)定氟喹諾酮藥物左氧氟沙星對(duì)臨床分離肺炎克雷伯菌的MIC和MPC,計(jì)算選擇指數(shù)(SI),選擇銅綠假單胞菌ATCC27853作為質(zhì)控菌株；鼻腔滴注法建立小鼠肺部肺炎克雷伯菌感染模型,比較不同接種劑量組死亡情況,確定最佳造模劑量；根據(jù)最終確定造模劑量建立小鼠肺部肺炎克雷伯菌感染模型,通過(guò)灌胃方法分別給予15mg/kg、30 mg/kg、60 mg/kg和90mg/kg左氧氟沙星治療,考察不同劑量組耐藥突變株的出現(xiàn)頻率及治療后MIC變化情況,評(píng)價(jià)不同劑量組與細(xì)菌耐藥突變株富集擴(kuò)增的關(guān)系；采用高效液相色譜法(HPLC)測(cè)定模型組小鼠治療一天、三天、五天、七天后的左氧氟沙星血藥濃度,參考相關(guān)PK/PD常數(shù),結(jié)合體外測(cè)定MPC和MSW值評(píng)價(jià)臨床應(yīng)用左氧氟沙星治療細(xì)菌性感染的劑量與療效關(guān)系。 結(jié)果 體外測(cè)定臨床分離肺炎克雷伯菌的MIC和MPC分別為0.0625μg／ml和1μg／ml,SI=16,根據(jù)CLSI藥敏標(biāo)準(zhǔn),此菌株為對(duì)左氧氟沙星敏感菌株；三個(gè)接種劑量組死亡率無(wú)明顯差異,綜合考慮接種時(shí)間、接種劑量等方面因素,確定最終接種濃度為108CFU/ml,接種劑量為30μl；給予不同劑量左氧氟沙星治療過(guò)程中,15mg/kg劑量組和90mg/kg劑量組,突變率較恒定,無(wú)明顯升高,MIC值多無(wú)變化,而30mg/kg和60mg/kg劑量組突變率明顯升高,MIC值升高的動(dòng)物數(shù)明顯增多,30mg/kg主要選擇低水平突變,MIC值較接種時(shí)升高2倍,而60mg/kg劑量組則主要選擇高水平突變,MIC值較接種時(shí)升高8倍；HPLC測(cè)定結(jié)果表明,在治療期間內(nèi),血藥濃度位于MSW內(nèi)的時(shí)間越長(zhǎng),細(xì)菌的突變率越高。 結(jié)論 不同劑量左氧氟沙星治療肺炎克雷伯菌感染的細(xì)菌突變率不同,不適當(dāng)劑量的藥物雖可以達(dá)到治療目的,卻會(huì)引起細(xì)菌突變株擴(kuò)增,在一定范圍內(nèi)水平傳播,加速細(xì)菌耐藥。根據(jù)本文的研究結(jié)果,推薦臨床應(yīng)用左氧氟沙星治療細(xì)菌性感染疾病時(shí),在兼顧安全的前提下采用大劑量沖擊療法。
[Abstract]:objective
In vitro determination of fluoroquinolone levofloxacin and minimum inhibitory concentration of clinical isolates of Klebsiella pneumoniae (Minimum Inhibitory, Concentration, MIC) and mutant prevention concentration (Mutant Prevention Concentration, MPO). The establishment of the lungs of mice infected with Klebsiella pneumoniae model, determination of pharmacokinetic / pharmacodynamic (PK/PD) parameters, to observe the drug the concentration of drug resistance mutations affect the enrichment degree of strain, combined with the determination results of experiments in vitro and in vivo animal preliminary validation of mutant selection window (MSW) theory, and the pharmacokinetic parameters of MSW theory and the evaluation of clinical levofloxacin in the treatment of pneumonia caused by Klebsiella pneumoniae dosing strategy.
Method
Using standard agar CLSI prescribed two times dilution method, determination of fluoroquinolone levofloxacin in clinical isolates of Klebsiella pneumoniae MIC and MPC, calculate the selection index (SI), selection of Pseudomonas aeruginosa ATCC27853 as control bacteria; nasal instillation method to establish mouse lung of Klebsiella pneumoniae infection model. The comparison of the death of different inoculation doses to determine the optimum dose; according to the final dose of mice lung Klebsiella pneumoniae infection model, 15mg/kg, were given by gavage for 30 mg/kg, 60 mg/kg and 90mg/kg of levofloxacin in the treatment of resistant strains of different dosage groups, investigated the frequency and changes of MIC after treatment the mutation, evaluation of different dose groups and bacteria resistant mutants were amplified by relationship enrichment; high performance liquid chromatography (HPLC) determination of model mice for a day, three days, five days, seven days After the levofloxacin blood concentration, refer to the relevant PK/PD constant, combined with MPC and MSW values in vitro to evaluate the relationship between the dose and efficacy of levofloxacin in the treatment of bacterial infections.
Result
Determination of clinical isolates of Klebsiella pneumoniae MIC and MPC were 0.0625 g / ml and 1 g / ml, SI=16 in vitro, drug sensitivity according to the CLSI standard, this strain is susceptible to levofloxacin strains; no significant differences between the three dose group mortality, considering the time of inoculation, inoculation amount etc. factors that determine the final inoculation concentration is 108CFU/ml, with the dose of 30 L; given different doses of levofloxacin in the treatment process, 15mg/kg and 90mg/kg groups, the mutation rate is relatively constant, significantly increased, MIC value had no change, while 30mg/kg and 60mg/kg dose group mutation rate was significantly increased, the MIC value of the number of animal elevated 30mg/kg increased significantly, the main choice of low level mutation, the MIC value is 2 times higher when inoculated, while the 60mg/kg group mainly choose high levels of mutation, the MIC value is vaccination increased 8 times; the results of HPLC analysis show that in the treatment period, blood concentration The longer the time in MSW, the higher the rate of mutation of bacteria.
conclusion
Different doses of levofloxacin in the treatment of Klebsiella pneumoniae infections in different mutation rate, improper drug dose can achieve the goal of treatment, but can cause bacterial mutant amplification and transmission within the certain range, accelerate the bacterial resistance. According to the results of this study, the recommended clinical application of levofloxacin in the treatment of bacterial infection disease and the impact of high-dose therapy on the premise of safety.

【學(xué)位授予單位】：中國(guó)醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R378

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