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目的研究耐青霉烯类肺炎克雷白杆菌(Carbapenem-Resistant Klebsiella Pneumoniae,CRKP)临床分布,及整合素1在CRKP多药耐药中的作用。方法在三年间从中国中部城市一家医院的多个科室采集唾液、血液及多种引流液临床标本,分离肺炎克雷白杆菌并鉴定CRKP。细菌药敏实验分析CRKP对13中抗生素的耐药情况。PCR扩增CRKP整合素1的可变区域,通过酶切及DNA测序技术分析整合素1可变区域的基因结构。结果共955例肺炎克雷白杆菌被分离出,其中CRKP共117例(12.3%)。2013,2014,2015年的Kp菌株中CRKP的分离率分别为8.9%(26/292),11.31%(38/336)及16.21%(53/327),呈逐步上升趋势。44.4%(52/117)的CRKP从ICU分离获得,61.5%(72/117)从痰液分离获得。117株CRKP中95%对氨苄西林/舒巴坦、氨曲南、亚胺培南、美罗培南、头孢他啶、头孢噻肟、头孢吡肟及哌拉西林耐药,但对替加环素和多粘菌素的耐药率较低,分别为12.8%和35.9%。CRKP中整合素1阳性检测率为77.8%(91/117),整合素1与妥布霉素,庆大霉素及阿米卡星这三种氨基糖苷类抗生素耐药密切相关(P<0.01)。整合素1可变区域基因盒检测显示,aad A2占64.84%(59/91),aac A4 cat B8-aad A1占23.1%(21/91),及aad A2-dfr A25占12.1%(11/91)。结论 CRKP有逐年增多的趋势,大部分同时具备多药耐药。整合素1介导的耐药基因盒在CRKP对氨基糖苷类抗生素耐药中发挥重要作用,其中aad A2是最常见的耐药基因。
Objective To investigate the clinical distribution of Carbapenem-Resistant Klebsiella Pneumoniae (CRKP) and the role of integrin-1 in CRKP multidrug resistance. Methods Three years clinical specimens of saliva, blood and various drainage fluids were collected from multiple departments of a hospital in the central city of China. Klebsiella pneumoniae was isolated and identified as CRKP. Antibiotic resistance of CRKP to 13 in bacterial susceptibility test was analyzed. The variable region of CRKP integrin 1 was amplified by PCR. The gene structure of integrin 1 variable region was analyzed by restriction enzyme digestion and DNA sequencing. Results A total of 955 cases of Klebsiella pneumoniae were isolated, of which 117 cases were CRKP (12.3%). The isolation rates of CRKP in Kp strains in 2013, 2014 and 2015 were 8.9% (26/292), 11.31% (38/336) and 16.21% (53/327), respectively, showing a gradual upward trend. 44.4% (52/117) of CRKP was isolated from the ICU and 61.5% (72/117) was isolated from sputum. 95% of 117 CRKP were resistant to ampicillin / sulbactam, aztreonam, imipenem, meropenem, ceftazidime, cefotaxime, cefepime and piperacillin but resistant to tigecycline and Colistin resistance rates were lower, respectively 12.8% and 35.9%. The positive rate of integrin-1 in CRKP was 77.8% (91/117). Integrin-1 was closely related to three aminoglycoside antibiotics, tobramycin, gentamicin and amikacin (P <0.01 ). Integrin-1 variable region gene cassette assay showed that aad A2 accounted for 64.84% (59/91), aac A4 cat B8-aad A1 accounted for 23.1% (21/91), and aad A2-dfr A25 accounted for 12.1% (11 / 91). Conclusion CRKP tends to increase year by year, most of which have multidrug resistance at the same time. Integrin-1-mediated resistance gene cassettes play an important role in the resistance of aminoglycoside antibiotics to CRKP, of which aad A2 is the most common resistance gene.