糖尿病足患者创面中耐碳青霉烯类鲍曼不动杆菌的生物膜基因和群体感应基因分析  

Analysis of biofilm genes and quorum sensing genes of carbapenem resistant Acinetobacter baumannii in the wounds of diabetic foot patients

作　　者：徐俊[1,2] 韩晓翠 何璐 冯书红[1] 孙东建[1] 曹晨 刘熙姣 张艳燕[2] 丁百兴 常柏[1] Xu Jun;Han Xiaocui;He Lu;Feng Shuhong;Sun Dongjian;Cao Chen;Liu Xijiao;Zhang Yanyan;Ding Baixing;Chang Bai(National Health Commission Key Laboratory of Hormones and Development,Tianjin Key Laboratory of Metabolic Diseases,Department of Diabetic Foot,Tianjin Medical University Chu Hsien-I Memorial Hospital&Institute of Endocrinology,Tianjin 300134,China;Department of Endocrinology/Rheumatology and Immunology,the Sixth Division Hospital,Xinjiang Production and Construction Corps,Wujiaqu 831300,China;Department of Infectious Medicine,the Sixth Division Hospital,Xinjiang Production and Construction Corps,Wujiaqu 831300,China;National Health Commission Key Laboratory of Hormones and Development,Tianjin Key Laboratory of Metabolic Diseases,Clinical Laboratory,Tianjin Medical University Chu Hsien-I Memorial Hospital&Institute of Endocrinology,Tianjin 300134,China;Institute of Antibiotics,Huashan Hospital,Fudan University,Shanghai 200040,China)

机构地区：[1]天津医科大学朱宪彝纪念医院糖尿病足科,天津市内分泌研究所,国家卫生健康委激素与发育重点实验室,天津市代谢性疾病重点实验室,天津300134 [2]新疆生产建设兵团第六师医院内分泌/风湿免疫科,五家渠831300 [3]新疆生产建设兵团第六师医院感染医学科,五家渠831300 [4]天津医科大学朱宪彝纪念医院检验科,天津市内分泌研究所,国家卫生健康委激素与发育重点实验室,天津市代谢性疾病重点实验室,天津300134 [5]复旦大学华山医院抗生素研究所,上海200040

出　　处：《中华烧伤与创面修复杂志》2024年第12期1166-1175,共10页Chinese Journal of Burns And Wounds

基　　金：国家自然科学基金面上项目(81973614);天津市医学重点学科(专科)建设项目(TJYXZDXK-032A)。

摘　　要：目的探讨糖尿病足患者创面中耐碳青霉烯类鲍曼不动杆菌(CRAB)携带生物膜基因和群体感应基因情况,以及CRAB对抗生素的耐药情况。方法该研究为回顾性观察性研究。2020年10月-2023年9月,天津医科大学朱宪彝纪念医院糖尿病足科收治177例符合入选标准的糖尿病足患者,其中男128例、女49例,年龄(56±10)岁。从前述患者的糖尿病足创面中培养出233株鲍曼不动杆菌,先用基质辅助激光解析电离飞行时间质谱鉴定细菌,然后采用动态比浊法通过全自动微生物系统做药物敏感试验分析耐药率。任意选择10株CRAB[来自10例患者,其中男9例、女1例,年龄(63±13)岁]和10株碳青霉烯类敏感鲍曼不动杆菌[CSAB,来自10例患者,其中男8例、女2例,年龄(63±9)岁],提取DNA并进行全基因组测序,使用综合抗生素耐药基因数据库对耐药基因进行注释,绘制系统发育树分析CRAB与CSAB的同源关系。将鲍曼不动杆菌的7个管家基因输入PubMLST网站,分析CRAB与CSAB的多位点序列分型。将测得的全部基因放入PubMLST网站查找每株鲍曼不动杆菌是否携带生物膜基因bap、csuA、csuB、csuA/B、csuC、csuD、csuE、pgaA、pgaB、pgaC、pgaD、bfmR、bfmS、ompA和群体感应基因abaI、abaR及鞭毛基因pilA,并比较CRAB与CSAB携带这些基因的差异。分析携带苯唑西林酶(OXA)耐药基因bla OXA的CRAB与CSAB所携带生物膜基因和群体感应基因情况。大体观察感染CRAB与CSAB的糖尿病足创面中是否存在胶冻样膜样结构,若存在则采用扫描电子显微镜观察其微观结构。结果检测到的鲍曼不动杆菌中,CSAB、CRAB、多重耐药鲍曼不动杆菌、泛耐药鲍曼不动杆菌的阳性检出率分别为16.7%(39/233)、83.3%(194/233)、95.3%(222/233)、34.3%(80/233),未检测到全耐药鲍曼不动杆菌。233株鲍曼不动杆菌对碳青霉烯类抗生素的耐药率均>80%;对头孢哌酮/舒巴坦的耐药率较低,为37%,对其他头孢类抗Objective To investigate the biofilm genes and quorum sensing genes of carbapenem resistant Acinetobacter baumannii(CRAB)in the wounds of diabetic foot patients.Methods This study was a retrospective observational study.The 233 strains of Acinetobacter baumannii were cultured from 177 inpatients(128 males and 49 females,aged(56±10)years)with diabetic foot admitted to the Department of Diabetic Foot of Tianjin Medical University Chu Hsien-I Memorial Hospital from October 2020 to September 2023.Two hundred and thirty-three Acinetobacter baumannii strains were detected by bacterial culture from the diabetic foot wounds of the aforementioned patients.All Acinetobacter baumannii strains were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,followed by analysis of their resistance rate using kinetic turbidimetric assay by a fully automated microbiological analysis system.Ten CRAB strains(from 10 patients,9 males and 1 female,aged(63±13)years)and 10 carbapenem sensitive Acinetobacter baumannii(CSAB;from 10 patients,8 males and 2 females,aged(63±9)years)strains were randomly selected,and the whole DNA genome was extracted and sequenced.The antibiotic resistance genes were annotated using a comprehensive antibiotic resistance gene database,and a phylogenetic tree was drawn to analyze the homologous relationship between CRAB and CSAB.The 7 housekeeping genes of Acinetobacter baumannii was entered into the PubMLST website to analyze the multi-locus sequence typing of CRAB and CSAB.All the measured genes were put into the PubMLST website to search for the biofilm genes bap,csuA,csuB,csuA/B,csuC,csuD,csuE,pgaA,pgaB,pgaC,pgaD,bfmR,bfmS,ompA carried by each Acinetobacter baumannii,as well as the quorum sensing genes abaI and abaR,and flagellar gene pilA.The differences in carrying these genes between CRAB and CSAB were compared.The biofilm genes and quorum sensing genes carried by CRAB and CSAB carrying oxacillinase(OXA)resistance gene bla OXA were analyzed.Gross observation was p

关 键 词：糖尿病足 生物膜 群体感应 鲍曼不动杆菌 碳青霉烯 抗生素耐药 

分 类 号：R587.2[医药卫生—内分泌]