泌尿道解葡萄糖苷棒状杆菌的分离、鉴定和药敏分析(1)
中圖分类号 R978.1 文献标志码 A 文章编号 1001-0408(2018)04-0496-05
DOI 10.6039/j.issn.1001-0408.2018.04.16
摘 要 目的：对临床分离的解葡萄糖苷棒状杆菌进行鉴定和药敏分析，为临床用药提供参考。方法：将从我院泌尿系统结石致尿路感染患者中段尿标本中分离得到的2株菌株接种至哥伦比亚血平板和麦康凯平板上，观察其生长情况并计数；采用基质辅助激光解析飞行时间质谱(MALDI-TOF-MS)检测菌株的蛋白质谱图；提取菌株DNA，采用聚合酶链反应扩增其16S核糖体RNA(rRNA)序列，对片段大小约为1 500 bp的目标条带进行双向测序，并与GenBank数据库进行Blast比对，鉴定菌种；采用Etest法监测2株菌株的耐药情况。结果：2株菌株在哥伦比亚血平板上生长(菌落数>105 CFU/mL)，在麦康凯平板上不生长；均为革兰氏阳性棒状杆菌，多呈栅栏样或八字形排列。MALDI-TOF-MS鉴定其均为解葡萄糖苷棒状杆菌，可信度为99.9%，在m/z 2 431、3 089、3 364、3 378、4 200、5 508、6 302、6 637、6 730、6 946、12 603处出现特征峰；Blast比对结果显示，2株菌株与GenBank中已知解葡萄糖苷棒状杆菌的序列同源性超过98%。药敏试验结果显示，菌株1对头孢曲松、环丙沙星耐药，对青霉素G等其他14种抗菌药物敏感；菌株2对头孢曲松、红霉素、环丙沙星、四环素、克林霉素耐药，对头孢噻肟中介，对其他10种抗菌药物敏感。结论：2株菌株均为解葡萄糖苷棒状杆菌，对常用抗菌药物的耐药性有所差异。该菌为罕见的泌尿道病原菌，且呈多重耐药，临床应根据菌株鉴定及药敏试验结果合理选用抗菌药物。
关键词 尿路感染；解葡萄糖苷棒状杆菌；基质辅助激光解析飞行时间质谱；鉴定；药敏试验
ABSTRACT OBJECTIVE： To identify and analyze drug sensitivity of Corynebacterium glucuronolyticum iscolated from clinic， and to provide reference for clinical drug use. METHODS： Two strains isolated from the urine specimens of urolithiasis-induced urinary tract infection patients in our hospital were inoculated into Columbia blood plate and the MacConkey plate. The growth of strains was observed and counted. Protein mass spectrometry of strains was detected by MALDI-TOF-MS. DNA of strains was extracted， and PCR was used to amplify the 16S ribosome RNA (rRNA) sequence. Bi-directional sequencing of 1 500 bp target bands was conducted. Blast comparison between it and GenBank database was conducted to identify bacterial strain. Drug resistance of 2 strains was monitored by Etest assay. RESULTS： Two strains grew on the Columbia blood plate (with colony forming unit >105 CFU/mL) and did not grow on the MacConkey plate. Two strains were Gram-positive Corynebacterium and showed palisading or eight type arrangement. Two strains were C. glucuronolyticum by MALDI-TOF-MS identification， with reliability of 99.9%. The characteristic peaks of m/z 2 431， 3 089， 3 364， 3 378， 4 200， 5 508， 6 302， 6 637， 6 730， 6 946， 12 603 appeared. Blast comparison showed that the sequence homology of 2 strains compared with C. glucuronolyticum strain known in GenBank were higher than 98%. Results of drug sensitivity test showed that strain 1 was resistant to ceftriaxone and ciprofloxacin， and sensitive to 14 other antibiotics as penicillin G； strain 2 was resistant to ceftriaxone， erythromycin， ciprofloxacin， tetracycline and clindamycin， moderately sensitive to cefotaxime， and sensitive to 10 other antibiotics. CONCLUSIONS： Two strains are C. glucuronolyticum， and drug resistance of them to commonly used antibiotics is different. The strains are rare pathogen of urinary tract and show multidrug resistance. Antibiotics should be selected according to the results of strain identification and drug sensitivity test., http://www.100md.com(韦柳华 罗国兰 李梦薇 林盛张 周格琛 邹燕)
DOI 10.6039/j.issn.1001-0408.2018.04.16
摘 要 目的：对临床分离的解葡萄糖苷棒状杆菌进行鉴定和药敏分析，为临床用药提供参考。方法：将从我院泌尿系统结石致尿路感染患者中段尿标本中分离得到的2株菌株接种至哥伦比亚血平板和麦康凯平板上，观察其生长情况并计数；采用基质辅助激光解析飞行时间质谱(MALDI-TOF-MS)检测菌株的蛋白质谱图；提取菌株DNA，采用聚合酶链反应扩增其16S核糖体RNA(rRNA)序列，对片段大小约为1 500 bp的目标条带进行双向测序，并与GenBank数据库进行Blast比对，鉴定菌种；采用Etest法监测2株菌株的耐药情况。结果：2株菌株在哥伦比亚血平板上生长(菌落数>105 CFU/mL)，在麦康凯平板上不生长；均为革兰氏阳性棒状杆菌，多呈栅栏样或八字形排列。MALDI-TOF-MS鉴定其均为解葡萄糖苷棒状杆菌，可信度为99.9%，在m/z 2 431、3 089、3 364、3 378、4 200、5 508、6 302、6 637、6 730、6 946、12 603处出现特征峰；Blast比对结果显示，2株菌株与GenBank中已知解葡萄糖苷棒状杆菌的序列同源性超过98%。药敏试验结果显示，菌株1对头孢曲松、环丙沙星耐药，对青霉素G等其他14种抗菌药物敏感；菌株2对头孢曲松、红霉素、环丙沙星、四环素、克林霉素耐药，对头孢噻肟中介，对其他10种抗菌药物敏感。结论：2株菌株均为解葡萄糖苷棒状杆菌，对常用抗菌药物的耐药性有所差异。该菌为罕见的泌尿道病原菌，且呈多重耐药，临床应根据菌株鉴定及药敏试验结果合理选用抗菌药物。
关键词 尿路感染；解葡萄糖苷棒状杆菌；基质辅助激光解析飞行时间质谱；鉴定；药敏试验
ABSTRACT OBJECTIVE： To identify and analyze drug sensitivity of Corynebacterium glucuronolyticum iscolated from clinic， and to provide reference for clinical drug use. METHODS： Two strains isolated from the urine specimens of urolithiasis-induced urinary tract infection patients in our hospital were inoculated into Columbia blood plate and the MacConkey plate. The growth of strains was observed and counted. Protein mass spectrometry of strains was detected by MALDI-TOF-MS. DNA of strains was extracted， and PCR was used to amplify the 16S ribosome RNA (rRNA) sequence. Bi-directional sequencing of 1 500 bp target bands was conducted. Blast comparison between it and GenBank database was conducted to identify bacterial strain. Drug resistance of 2 strains was monitored by Etest assay. RESULTS： Two strains grew on the Columbia blood plate (with colony forming unit >105 CFU/mL) and did not grow on the MacConkey plate. Two strains were Gram-positive Corynebacterium and showed palisading or eight type arrangement. Two strains were C. glucuronolyticum by MALDI-TOF-MS identification， with reliability of 99.9%. The characteristic peaks of m/z 2 431， 3 089， 3 364， 3 378， 4 200， 5 508， 6 302， 6 637， 6 730， 6 946， 12 603 appeared. Blast comparison showed that the sequence homology of 2 strains compared with C. glucuronolyticum strain known in GenBank were higher than 98%. Results of drug sensitivity test showed that strain 1 was resistant to ceftriaxone and ciprofloxacin， and sensitive to 14 other antibiotics as penicillin G； strain 2 was resistant to ceftriaxone， erythromycin， ciprofloxacin， tetracycline and clindamycin， moderately sensitive to cefotaxime， and sensitive to 10 other antibiotics. CONCLUSIONS： Two strains are C. glucuronolyticum， and drug resistance of them to commonly used antibiotics is different. The strains are rare pathogen of urinary tract and show multidrug resistance. Antibiotics should be selected according to the results of strain identification and drug sensitivity test., http://www.100md.com(韦柳华 罗国兰 李梦薇 林盛张 周格琛 邹燕)
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