纸片扩散法对OXA型耐碳青霉烯类鲍氏不动杆菌的鉴定作用  

Identification of OXA-type carbapenem-resistant Acinetobacter baumannii with disk diffusion method

作　　者：王艳侠[1] 周战修[1] 蔡辉[1] 陈露露 姜梅杰[3] 

机构地区：[1]山东省济宁医学院附属金乡医院检验科,山东济宁272200 [2]济宁市金乡县疾病预防与控制中心检验科,山东济宁272200 [3]泰安市中心医院检验科,山东泰安271000

出　　处：《中华医院感染学杂志》2016年第6期1219-1222,共4页Chinese Journal of Nosocomiology

基　　金：山东省自然科学基金资助项目(22011HM036);济宁市科技计划基金资助项目(2014jnjc18)

摘　　要：目的探索简便易行的纸片扩散法,用于临床实验室OXA型耐碳青霉烯类鲍氏不动杆菌的筛选。方法收集济宁医学院附属金乡医院2012年1月-2014年6月耐碳青霉烯类鲍氏不动杆菌32株,利用PCR筛出其中的OXA各亚型并将其编号纳入试验组;选取无菌水、1株鲍氏不动杆菌ATCC17978、3株IMP型和3株VIM型耐碳青霉烯鲍氏不动杆菌设为4个对照组;将金黄色葡萄球菌涂在M-H平板上并贴好苯唑西林纸片;调整各组0.5麦氏单位菌液,并取10μl加入到苯唑西林纸片上;将M-H平板放置于培养箱35℃孵育48h并观察苯唑西林纸片抑菌圈内菌落生长情况。结果 PCR筛选出OXA型耐碳青霉烯类鲍氏不动杆菌25株纳入试验组,培养12h后,5组苯唑西林纸片周围均形成清晰的抑菌圈,抑菌圈内无菌落生长;培养24h后,试验组抑菌圈内有少数菌落生长;培养36h后,试验组抑菌圈内菌落变大且数量变多;继续培养至48h,试验组抑菌圈菌落无明显生长;整个培养过程中,4个对照组抑菌圈内均无菌落生长。结论试验所应用的纸片扩散法可筛选出产OXA型耐碳青霉烯类鲍氏不动杆菌,且最佳观察时间为35℃孵育36h。OBJECTIVE To explore the use of disk diffusion method in clinical laboratory screening of the OXA-type carbapenem-resistant Acinetobacter baumannii.METHODS A total of 32 strains of carbapenem-resistant A.baumannii were collected from the Affiliated Jinxiang Hospital of Jining Medical College from Jan 2012 to Jun 2014;the OXA-subtypes strains were screened out by using PCR and assigned as the test group;the sterile water,1strain of A.baumannii ATCC17978,3strains of IMP-type,and 3strains of VIM-type carbapenem-resistant A.baumannii were set as four control groups.The oxacillin disc was put on the MH plate after the Staphylococcus aureus was coated,then the bacteria concentrations of all the groups were diluted to 0.5Maxwell units,and 10μL of the diluted bacteria solution was added to the oxacillin disc,then the M-H plates were incubated at 35℃for 48 hours,and the growth of bacterial colony in the oxacillin bacteriostatic circle was observed.RESULTS Totally 25 strains of OXA-type carbapenem-resistant A.baumannii were screened out by using the PCR and were assigned as the test group,the clear inhibition zones were observed around the oxacillin disk in the 5groups after the culture for 12 hours,and there was no growth of bacterial colony in the bacteriostatic circle.There was growth of a few of bacterial colonies in the bacteriostatic circle of the test group after the culture for 24 hours.The bacterial colonies in the bacteriostatic circle grew bigger and were increased in the test group after the culture for 36 hours.There was no clear growth of bacterial colony in the bacteriostatic circle of the test group after the culture for 48 hours.There was no growth of bacterial colony in any of the four control groups during the whole process of culture.CONCLUSION The disk diffusion method can screen out the OXA-type carbapenem-resistant A.baumannii strains,and the optimal observation time is at the 36 hours of incubation under 35℃.

关 键 词：纸片扩散法 鲍氏不动杆菌 碳青霉烯酶 

分 类 号：R378.99[医药卫生—病原生物学]