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目的:分析惠东地区肠杆菌科细菌中超广谱β-内酰胺酶类型及外膜蛋白缺失情况。方法:收集2013年1月至2013年12月期间惠东地区各医院,各科室血、尿、痰、大便、穿刺液、分泌物、胆汁等样本中分离培养的菌株,依据2010年版CLSI文件M100-S20判断依据确证阳性菌株246株。本研究采用荧光定量PCR反应检测细菌基因型,采用琼脂稀释法及E-tesr法测定MIC值,对部分耐药现象异常的菌株采用SDS-PAGE凝胶电泳研究外膜蛋白结构。结果:共有270株肠杆菌科细菌检测出β-内酰胺酶,检出率为95.74%。在大肠埃希菌与变异杆菌中,CTX-M-9组基因检出率明显高于CTX-M-1组基因(P<0.01),在肺炎克雷伯菌中CTX-M-1组基因检出率明显高于CTX-M-9组基因(P<0.01)。282株产ESBLs的菌株对头孢唑啉、头孢噻肟、头孢曲松株均呈耐药性。然而93.33%、95.56%的奇异变形杆菌对头孢他啶及氨曲南敏感。提取对头孢唑啉、头孢噻肟、头孢曲松株耐药,却对氨曲南十分敏感的菌株的外膜蛋白进行SDS-PAGE电泳,未发现明显的结构缺失。结论:在惠东地区,肠杆菌科细菌普遍能够产生ESBLs,故临床进行治疗时应根据细菌种类及基因型确定治疗方案。
OBJECTIVE: To analyze the type of extended-spectrum β-lactamase and the deletion of outer membrane protein in Enterobacteriaceae in Huidong area. Methods: The strains isolated from blood, urine, sputum, stool, puncture fluid, secretions and bile in hospitals and departments of Huidong area from January 2013 to December 2013 were collected and analyzed according to CLSI document M100 -S20 According to the confirmation of positive 246 strains. In this study, bacterial genotypes were detected by real-time PCR. MIC values were determined by agar dilution method and E-tesr method. SDS-PAGE gel electrophoresis was used to study the outer membrane protein structure of some strains with abnormal drug resistance. Results: A total of 270 Enterobacteriaceae bacteria detected β-lactamases, the detection rate was 95.74%. The gene detection rate of CTX-M-9 group was significantly higher than that of CTX-M-1 group (P <0.01) in Escherichia coli and Mutant bacilli. In the CTX-M-1 group of Klebsiella pneumoniae, The detection rate was significantly higher than that of CTX-M-9 group (P <0.01). 282 strains of ESBLs-producing strains of cefazolin, cefotaxime, ceftriaxone strains were resistant. However, 93.33% and 95.56% Proteus mirabilis were sensitive to ceftazidime and aztreonam. The outer membrane proteins of strains resistant to cefazolin, cefotaxime and ceftriaxone, but sensitive to aztreonam, were extracted by SDS-PAGE and no obvious structural loss was found. Conclusion: Enterobacteriaceae bacteria generally produce ESBLs in Huidong area. Therefore, the clinical treatment should be based on the type of bacteria and genotype to determine the treatment.