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目的:研究磷霉素联合5种中药单、复方对产KPC肠杆菌科细菌体外药敏实验的抑菌效果。方法:2012年7月~2013年6月从杭州市中医院分离到的20株碳青霉烯不敏感的肠杆菌科细菌,其中有11株肺炎克雷伯菌,7株大肠埃氏菌,2株摩根摩根菌。改良Hodge实验及特异性PCR扩增和序列分析细菌KPC碳青霉烯酶的产生情况。琼脂稀释法分别测定亚胺培南、厄他培南、磷霉素、大黄、黄芪、黄连、金银花、鱼腥草及磷霉素联合中药单复方对待检菌的最小抑菌浓度(MIC)。结果:20株对碳青霉烯不敏感的肠杆菌科细菌均产KPC-2型碳青霉烯酶。体外药敏试验显示敏感率,亚胺培南为5%(1/20)、厄他培南为10%(2/20)、磷霉素为65%(13/20)(根据EUCAST标准≤32μg/mL);中药中黄芪的抑菌效果最好,MIC为25~50 mg/mL,黄连其次,MIC为50~100 mg/mL;大黄、鱼腥草、金银花抑菌效果最差,但金银花、金银花加黄连黄芪复方、鱼腥草加黄连黄芪复方对摩根摩根菌抑菌效果明显,MIC为12.5~25 mg/mL;磷霉素联合中药后,除高度耐药株外,大部分菌株MIC值均降低了1~3个稀释度。尤其是磷霉素抗菌活性不佳的摩根摩根菌株,联合除鱼腥草外其他中药后磷霉素的MIC下降可达6个稀释度。结论:通过琼脂稀释法测定的中药抑菌效果有限且与菌株是否产KPC酶无关。联合中药抗碳青霉烯不敏感的肠杆菌细菌感染可降低磷霉素MIC,提高药物效能。
OBJECTIVE: To study the bacteriostatic effect of fosfomycin combined with five kinds of traditional Chinese medicine alone and in combination on the susceptibility test of bacterium producing KPC Enterobacteriaceae in vitro. METHODS: Twenty carbapenem insensitive Enterobacteriaceae isolated from Hangzhou Hospital of Traditional Chinese Medicine from July 2012 to June 2013 included 11 strains of Klebsiella pneumoniae, 7 strains of Escherichia coli, 2 strains of Morgan bacteria. Improved Hodge experiment and specific PCR amplification and sequence analysis of bacterial KPC carbapenemase production. Agar dilution method were determined imipenem, ertapenem, fosfomycin, rhubarb, Astragalus, Coptis, honeysuckle, Houttuynia and fosfomycin combined with Chinese medicine single prescriptions for antibacterial MIC (minimum inhibitory concentration). Results: 20 strains of Enterobacteriaceae insensitive to carbapenem produced KPC-2 carbapenemase. In vitro susceptibility tests showed a sensitivity of 5% (1/20) for imipenem, 10% (2/20) for ertapenem, and 65% (13/20) for fosfomycin (EUCAST ≤ 32μg / mL). Astragalus membranaceus had the best antibacterial activity with the MIC of 25-50 mg / mL, followed by Coptis chinensis Franch and MIC of 50-100 mg / mL. Rhubarb, Houttuynia, Honeysuckle, honeysuckle plus Huanglian Astragalus compound, Houttuynia, Coptis and Astragalus compound on the Morgan root inhibitory effect was obvious, MIC 12.5 ~ 25 mg / mL; fosfomycin combined with traditional Chinese medicine, in addition to highly resistant strains, the majority of strains MIC Values were reduced by 1 to 3 dilutions. Especially fosfomycin Morgan antibacterial activity of poor Morgan strains, combined with other than Houttuynia outside the fosfomycin MIC decline of up to 6 dilutions. Conclusion: The antibacterial effect of traditional Chinese medicine determined by agar dilution method is limited and it has nothing to do with the production of KPC enzyme. Combination of anti-carbapenem-resistant Enterobacteriaceae bacterial infection can reduce the fosfomycin MIC, improve drug efficacy.