非发酵菌碳青霉烯酶检测及分子流行病学研究.docVIP

非发酵菌碳青霉烯酶检测及分子流行病学研究 谷秀梅1，杨敏2，刘文恩(1，彭婉婵1，李艳明1，李虹玲1 【摘要】目的 调查湘雅医院临床分离碳青霉烯类耐药非发酵菌产碳青霉烯酶的情况，探讨产酶菌株的分子流行病学特点及可移动遗传元件分布。方法 收集湘雅医院检验科临床微生物室2011年10月-2012年3月临床各类标本中非重复碳青霉烯类不敏感非发酵菌，根据CLSI 2010年标准采用纸片扩散法进行药敏试验；采用2种改良Hodge试验进行碳青霉烯酶表型确证；聚合酶链式反应（PCR）检测主要碳青霉烯酶基因blaKPC、blaIMP、blaVIM-1、blaVIM-2、blaSPM、blaNDM-1、blaOXA-23、blaOXA-40、和blaOXA-58，对 PCR扩增阳性产物进行基因测序以确定基因型别；对碳青霉烯酶基因扩增阳性菌株再进行4类可移动遗传元件如整合子遗传标记( intI 1、intI 2、intI 3)、转座子遗传标记( merA、tnpU、tnp513、tnpA) 、插入序列遗传标记（ISAba1、IS26）及接合性质粒遗传标记（traA、trbC）Detection of carbapenemases and analysis epidemiology in nonfermenters GU Xiu-mei1 , YANG Min2, LIU Wen-en1 , PENG Wan-chan1, LI Yan-ming1，LI Hong-ling1 Corresponding author :LIU Wen-en ,E-mail:liuwenen@. 【Abstract】Objectives To investigate the drug resistance and prevalence of carbapenemases in nonfermentative clinical isolates collected from our hospital and to study molecular epidemiology and resistance mechanism of carbapenemase-producing isolates. Methods Nonrepetitive carbapenem-resistant nonfermenters were collected in the clinical microbiology laboratory in our hospital from October 2011 to March 2012. Nonfermenters resistant to carbapenems were screened by disk diffusion method. Two kinds of Modified Hodge Test were conducted to detect the carbapenemase-producing isolates. The blaKPC, blaIMP, blaVIM-1, blaVIM-2, blaSPM, blaNDM-1, blaOXA-23, blaOXA-40, blaOXA-58 genes were detected by specific PCR, and the amplified products were sequenced. . PCR was used to amplify four kinds of mobile genetic elements such as integron genetic marks (intI 1,intI 2 and intI 3),transposon genetic marks ( merA,tnpU,tnp513 and tnpA)、insertion sequence genetic marks (ISAba1IS26) and conjunction plasmid genetic marks (traA and trbC)) of carbapenemase-producing isolates. Results In the 82 nonfermentative clinical isolates, resistant rates to the major antibiotics were higher than 60%. The resistant rates of the 27 Pseudomonas aeruginosa isolates to amikacin ，ceftazidime，and cetriaxone were 22.2% 、37.0%