多重耐药性铜绿假单胞菌的分离鉴定及小鼠烧伤感染模型的建立  

Isolation and identification of multi-drug resistant Pseudomanas areuginosa and burned mice infection model construction

作　　者：郝羽翀 刘洋洋 王亚利 史红艳[1] HAO Yu-chong LIU Yang yang WANG Ya-li et al.(The Basic Medical School of J ilin University, Changchun 130021 ,China)

机构地区：[1]吉林大学基础医学院,吉林长春130021

出　　处：《中国实验诊断学》2017年第9期1622-1625,共4页Chinese Journal of Laboratory Diagnosis

摘　　要：目的分离鉴定多重耐药性铜绿假单胞菌,制备铜绿假单胞菌感染的小鼠烫伤模型。方法常规方法分离纯化铜绿假单胞菌,并经16sRNA序列分析鉴定铜绿假单胞菌;平板稀释法测定分离菌株对18种抗菌药物的耐药性;应用烫伤仪确定小鼠对100℃下不同烫伤面积和烫伤时间的耐受情况,确定最佳烫伤时间和面积建立小鼠烫伤模型;对小鼠烫伤模型皮下感染不同浓度的铜绿假单胞菌,确定72h内全部小鼠死亡的细菌最小致死量,最小致死量感染组小鼠死亡后立即应用平板稀释法检测肝和脾内细菌荷载量。结果分离鉴定得到5株多重耐药性铜绿假单胞菌,其中耐药种类最多的为PA53,该菌对检测的18种抗菌药物中的14种耐药;烫伤温度100℃,面积2.5cm2,烫伤时间10s可获得稳定的小鼠烫伤模型;PA53对小鼠烫伤模型感染的最小致死量为1.8×1011 CFU/ml,250μl;死亡小鼠肝和脾内的细菌荷载量分别为108PFU/g和1010 PFU/g数量级。结论获得多重耐药性铜绿假单胞菌PA53;成功构建小鼠烫伤模型,并测定了PA53对烫伤模型小鼠的最小致死量及死亡小鼠部分脏器的细菌荷载量。Objective In order to isolate and identify the multi-drug resistant Pseudomonas aeruginosa（MDRPA） and rebuilt infection model of burned-ice by MDRPA. Methods The burned-mice model was rebuilt by burning instrument according to different area and time at 100℃. Pseudomonas aeruginosa was isolated and purified by the conventional method,and identified by 16sRNA sequence analysis ; The 18 kinds of anti-bacterial drugs were administrated to MDRPA by plate dilution method; Different concentrations of MDRPA were infected by subcutaneous injection to burned-mice model to measure the 100% minimum lethal dose within 72h; And the loading bacteria of spleens and livers of the dead mice in 100% minimum lethal dose infection group were tested by plate dilution method. Results The multi-drug resistant Pseudomonas aeruginosa PAS3 was selected finally,this bacterial is resistant to 14 kinds of antibiotics of all 18. The burn area and burn time were 2.5 cm^2 , 10 s. The minimum lethal dose of PA53 to burned mice was 1.8× 10^11CFU/ml,250μl; The concentration of PA53 in liver and spleen were10^8PFU/g and10^10PFU/g respectively. Conclusion A multi-drug resistant Pseudomonas aeruginosa PA53 was isolated and identified,and the burned-mice in fection model was successfully constructed. The minimal lethal dose of PA53 and the bacterial loading of spleen and liver of dead mice were measured.

关 键 词：多重耐药性 铜绿假单胞菌 烧伤模型 最小致死量 

分 类 号：R-332[医药卫生] R644