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：《中國醫(yī)科大學(xué)》2010年碩士論文　論文類型：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 體外測定氟喹諾酮類藥物左氧氟沙星對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,測定相關(guān)藥代／藥效動力學(xué)(PK/PD)參數(shù),觀察藥物濃度對耐藥突變菌株富集程度的影響,結(jié)合體外實驗測定結(jié)果,在動物體內(nèi)初步驗證突變選擇窗(MSW)理論,并通過MSW理論與已有的藥代動力學(xué)參數(shù),對臨床左氧氟沙星治療肺炎克雷伯菌肺炎的給藥策略進行評價。 方法 采用CLSI規(guī)定的標準瓊脂平皿二倍稀釋法,測定氟喹諾酮藥物左氧氟沙星對臨床分離肺炎克雷伯菌的MIC和MPC,計算選擇指數(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,評價不同劑量組與細菌耐藥突變株富集擴增的關(guān)系；采用高效液相色譜法(HPLC)測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)合體外測定MPC和MSW值評價臨床應(yīng)用左氧氟沙星治療細菌性感染的劑量與療效關(guān)系。 結(jié)果 體外測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藥敏標準,此菌株為對左氧氟沙星敏感菌株；三個接種劑量組死亡率無明顯差異,綜合考慮接種時間、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程中,15mg/kg劑量組和90mg/kg劑量組,突變率較恒定,無明顯升高,MIC值多無變化,而30mg/kg和60mg/kg劑量組突變率明顯升高,MIC值升高的動物數(shù)明顯增多,30mg/kg主要選擇低水平突變,MIC值較接種時升高2倍,而60mg/kg劑量組則主要選擇高水平突變,MIC值較接種時升高8倍；HPLC測定結(jié)果表明,在治療期間內(nèi),血藥濃度位于MSW內(nèi)的時間越長,細菌的突變率越高。 結(jié)論 不同劑量左氧氟沙星治療肺炎克雷伯菌感染的細菌突變率不同,不適當(dāng)劑量的藥物雖可以達到治療目的,卻會引起細菌突變株擴增,在一定范圍內(nèi)水平傳播,加速細菌耐藥。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菌性感染疾病時,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】：中國醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R378

【參考文獻】

相關(guān)期刊論文 前10條

1 胡靜;姚云清;傅靜亦;夏云;左儀;;2005～2007年醫(yī)院病原菌分布和耐藥性分析[J];重慶醫(yī)學(xué);2009年07期

2 聶大平;董楓;石宏宴;;左氧氟沙星、環(huán)丙沙星單用和聯(lián)合其他抗菌藥物對銅綠假單胞菌防突變濃度的研究[J];中國感染控制雜志;2007年06期

3 方正;修清玉;黃海;陳吉泉;何禮賢;萬歡英;周新;;注射用左氧氟沙星治療急性細菌性感染的多中心隨機對照研究[J];中國抗感染化療雜志;2005年06期

4 曹國英;張菁;郁繼誠;施耀國;;HPLC法測定人血漿中左氧氟沙星濃度[J];中國臨床藥學(xué)雜志;2008年02期

5 田俊生;楊佳鳳;王躍飛;潘桂湘;高秀梅;張恒昆;;HPLC法測定健康人體血漿中左氧氟沙星濃度[J];天津中醫(yī)藥大學(xué)學(xué)報;2007年02期

6 崔俊昌;劉又寧;王睿杭;童衛(wèi)杭;李朝霞;;左氧氟沙星對兔組織籠金黃色葡萄球菌感染的療效研究[J];中國臨床藥理學(xué)與治療學(xué);2007年03期

7 李朝霞;劉又寧;王睿;童衛(wèi)杭;程仕虎;;左氧氟沙星聯(lián)合萬古霉素縮小金黃色葡萄球菌耐藥突變選擇窗的初步研究[J];中國臨床藥理學(xué)與治療學(xué);2007年08期

8 崔俊昌;劉又寧;王睿;童衛(wèi)杭;梁蓓蓓;;左氧氟沙星藥代動力學(xué)/藥效動力學(xué)參數(shù)與金黃色葡萄球菌耐藥的相關(guān)性研究[J];中國臨床藥理學(xué)與治療學(xué);2007年09期

9 黃瀚;劉世坤;;對抗菌藥物防突變濃度及突變選擇窗兩概念的探討與思索[J];中國臨床藥理學(xué)與治療學(xué);2008年01期

10 鐘大放;以加權(quán)最小二乘法建立生物分析標準曲線的若干問題[J];藥物分析雜志;1996年05期

，

本文編號：1402324