检验医学
檢驗醫學
검험의학
LABORATORY MEDICINE
2014年
11期
1178-1183
,共6页
曾章锐%王卫萍%黄梅%邵海枫
曾章銳%王衛萍%黃梅%邵海楓
증장예%왕위평%황매%소해풍
铜绿假单胞菌%头孢吡肟%PSE-1%KPC%外排系统
銅綠假單胞菌%頭孢吡肟%PSE-1%KPC%外排繫統
동록가단포균%두포필우%PSE-1%KPC%외배계통
Pseudomonas aeruginosa%Cefepime%PSE-1%KPC%Efflux system
目的：探讨临床分离的铜绿假单胞菌对头孢吡肟（FEP）敏感性低于头孢他啶（CAZ）的原因。方法收集临床分离的经 Vitek 2 Compact 全自动微生物分析系统检测 PEF 最小抑菌浓度（MIC）高于 CAZ MIC 的铜绿假单胞菌60株，琼脂稀释法复查 FEP 和 CAZ 的 MIC 值，聚合酶链反应（PCR）扩增耐药基因，实时荧光定量 PCR分析细菌外排系统表达情况。结果手工琼脂稀释法检测与 Vitek 2 Compact 全自动微生物分析系统检测存在5％的差别，PCR 扩增发现部分菌株存在编码 KPC 和 PSE-1的耐药基因，外排系统表达显示以 mexB 和 mexD 为主。测序结果发现 mexB 的3个调控基因只有 nalC Gly70→Glu（GGG→GAG）改变，其他2组调控基因未发现有意义的突变，mexD 外排系统过度表达的调控基因出现 nfxB Gly109→Val（GGC→GTC）。结论FEP 敏感性低于CAZ 的主要原因可能是编码 KPC 与 PSE-1的耐药基因的存在和外排系统 mexAB-OprM、mexCD-OprJ 表达增加共同作用的结果。
目的：探討臨床分離的銅綠假單胞菌對頭孢吡肟（FEP）敏感性低于頭孢他啶（CAZ）的原因。方法收集臨床分離的經 Vitek 2 Compact 全自動微生物分析繫統檢測 PEF 最小抑菌濃度（MIC）高于 CAZ MIC 的銅綠假單胞菌60株，瓊脂稀釋法複查 FEP 和 CAZ 的 MIC 值，聚閤酶鏈反應（PCR）擴增耐藥基因，實時熒光定量 PCR分析細菌外排繫統錶達情況。結果手工瓊脂稀釋法檢測與 Vitek 2 Compact 全自動微生物分析繫統檢測存在5％的差彆，PCR 擴增髮現部分菌株存在編碼 KPC 和 PSE-1的耐藥基因，外排繫統錶達顯示以 mexB 和 mexD 為主。測序結果髮現 mexB 的3箇調控基因隻有 nalC Gly70→Glu（GGG→GAG）改變，其他2組調控基因未髮現有意義的突變，mexD 外排繫統過度錶達的調控基因齣現 nfxB Gly109→Val（GGC→GTC）。結論FEP 敏感性低于CAZ 的主要原因可能是編碼 KPC 與 PSE-1的耐藥基因的存在和外排繫統 mexAB-OprM、mexCD-OprJ 錶達增加共同作用的結果。
목적：탐토림상분리적동록가단포균대두포필우（FEP）민감성저우두포타정（CAZ）적원인。방법수집림상분리적경 Vitek 2 Compact 전자동미생물분석계통검측 PEF 최소억균농도（MIC）고우 CAZ MIC 적동록가단포균60주，경지희석법복사 FEP 화 CAZ 적 MIC 치，취합매련반응（PCR）확증내약기인，실시형광정량 PCR분석세균외배계통표체정황。결과수공경지희석법검측여 Vitek 2 Compact 전자동미생물분석계통검측존재5％적차별，PCR 확증발현부분균주존재편마 KPC 화 PSE-1적내약기인，외배계통표체현시이 mexB 화 mexD 위주。측서결과발현 mexB 적3개조공기인지유 nalC Gly70→Glu（GGG→GAG）개변，기타2조조공기인미발현유의의적돌변，mexD 외배계통과도표체적조공기인출현 nfxB Gly109→Val（GGC→GTC）。결론FEP 민감성저우CAZ 적주요원인가능시편마 KPC 여 PSE-1적내약기인적존재화외배계통 mexAB-OprM、mexCD-OprJ 표체증가공동작용적결과。
Objective To investigate the reasons of less susceptibility mechanism to cefepime (FEP)than to ceftazidime (CAZ)in clinical isolates of Pseudomonas aeruginosa.Methods A total of 60 isolates of Pseudomonas aeruginosa which had been tested being less susceptible to FEP than to CAZ for minimal inhibitory concentration (MIC) by Vitek 2 Compact automatic microorganism analyzer system were collected.Agar dilution method was used to reexamine the MIC of FEP and CAZ.The resistance genes were amplified by polymerase chain reaction (PCR).The expressions of efflux system were analyzed by real-time fluorescence quantitation PCR.Results The 5% of error rate was showed between agar dilution method and Vitek 2 Compact system.There were isolates with KPC and PSE-1 resistance genes by PCR amplification.There were mainly the expressions of mexB and mexD in efflux system.The sequencing of regulatory genes showed that there were 3 isolates for mexB which the Gly70(GGG)of nalC was substitute for Glu(GAG)(GGG→GAG)and 2 isolates for mexD which the Gly1 09(GGC)of nfxB was substitute for Val (GTC) (GGC→GTC).Conclusions Less susceptibility to FEP than to CAZ in clinical isolates of Pseudomonas aeruginosa is due to the production of KPC and PSE-1 resistance genes and the increasing expression levels of mexAB-OprM and mexCD-OprJ.
