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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 a simple and convenient disk diffusion method for the screening of OXA carbapenem resistant Acinetobacter baumannii in clinical laboratory. Methods Thirty-two strains of carbapenem-resistant Acinetobacter baumannii were collected from Jinxiang Hospital affiliated to Jining Medical College from January 2012 to June 2014. The OXA subtypes were screened by PCR and included in the experimental group. Select sterile water, Acinetobacter baumannii ATCC17978, 3 IMP and 3 VIM carbapenem-resistant Acinetobacter baumannii was set as 4 control groups; Staphylococcus aureus coated MH plate And attached oxacillin paper; adjust each group 0.5 Macrobrachium unit bacilli, and take 10μl added to the oxacillin paper; the MH plate was incubated in an incubator at 35 ℃ 48h and observed oxacillin paper antibacterial Circle the growth of the colony. Results 25 strains of OXA carbapenem-resistant Acinetobacter baumannii were screened out by PCR. After cultured for 12h, all the 5 groups of oxacillin disks formed a clear inhibition zone around them, ; After cultured for 24h, there were a few colonies in the zone of inhibition in the experimental group; after 36h of culture, the number of colonies in the zone of inhibition zone became larger and more in the experimental group; no significant growth was observed in the zone of inhibition zone in the experimental group after 48h incubation; In the process of culture, there was no colony growth in the four control groups. Conclusion The OXA type carbapenem-resistant Acinetobacter baumannii was screened by the disk diffusion method. The optimal observation time was 35 ℃ for 36 hours.