中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2013年
6期
416-420
,共5页
胡丽庆%王盛%史煜波%翁幸鐾
鬍麗慶%王盛%史煜波%翁倖鐾
호려경%왕성%사욱파%옹행폐
奇异变形杆菌%blaKPC%PCR%质粒%电转化
奇異變形桿菌%blaKPC%PCR%質粒%電轉化
기이변형간균%blaKPC%PCR%질립%전전화
Proteus mirabilis%blaKPC%PCR%Plasmid%Electroporation
目的 研究本地区近4年临床分离的非重复碳青霉烯类耐药的奇异变形杆菌可能存在的碳青霉烯类酶基因型别.方法 回顾分析2009到2012年临床分离奇异变形杆菌的资料筛选出非重复的碳青霉烯类抗生素耐药的菌株15株；用浓度梯度法(E-试条法)检测其对亚胺培南(IPM)、美罗培南(MEM)、厄他培南(ETP)、环丙沙星(CIP)、阿米卡星(AK)和米诺环素(MIN)的MIC值；Hodge试验进行产碳青霉烯酶的确认,同时对试验菌株进行耐药基因的PCR扩增检测及测序,对阳性扩增菌株的质粒提取物与叠氮钠耐受的敏感态大肠埃希菌J53(E.coli J53)进行电转化,并对电转化后的菌株进行目标基因的PCR扩增及碳青霉烯类抗生素MIC值的检测.结果 2009到2012年,筛选到15株耐碳青霉烯类抗生素耐药的菌株；IPM、MEM、ETP的耐药率均为100％,CIP的耐药率为86.7％(13/15),AK的耐药率为33.3％ (5/15),MIN的耐药率为80％(12/15)；15株细菌中Hodge试验阳性7株,blaKPC基因阳性11株,经测序检测确认为blaKPC-2型；11株blaKPC基因阳性菌株的质粒提取物经电转化大肠埃希菌J53,使转化子大肠埃希菌J53对亚胺培南、美洛培南和厄他培南的MIC值增高2～64倍,PCR扩增转化子大肠埃希菌J53 blaKPC基因均阳性.结论 本地区对碳青霉烯类抗生素耐药的奇异变形杆菌,对阿米卡星的耐药率相对较低.其碳青霉烯酶的基因型主要为位于质粒上的blaKPC-2基因型,易于在菌株间流行,临床应引起高度重视.
目的 研究本地區近4年臨床分離的非重複碳青黴烯類耐藥的奇異變形桿菌可能存在的碳青黴烯類酶基因型彆.方法 迴顧分析2009到2012年臨床分離奇異變形桿菌的資料篩選齣非重複的碳青黴烯類抗生素耐藥的菌株15株；用濃度梯度法(E-試條法)檢測其對亞胺培南(IPM)、美囉培南(MEM)、阨他培南(ETP)、環丙沙星(CIP)、阿米卡星(AK)和米諾環素(MIN)的MIC值；Hodge試驗進行產碳青黴烯酶的確認,同時對試驗菌株進行耐藥基因的PCR擴增檢測及測序,對暘性擴增菌株的質粒提取物與疊氮鈉耐受的敏感態大腸埃希菌J53(E.coli J53)進行電轉化,併對電轉化後的菌株進行目標基因的PCR擴增及碳青黴烯類抗生素MIC值的檢測.結果 2009到2012年,篩選到15株耐碳青黴烯類抗生素耐藥的菌株；IPM、MEM、ETP的耐藥率均為100％,CIP的耐藥率為86.7％(13/15),AK的耐藥率為33.3％ (5/15),MIN的耐藥率為80％(12/15)；15株細菌中Hodge試驗暘性7株,blaKPC基因暘性11株,經測序檢測確認為blaKPC-2型；11株blaKPC基因暘性菌株的質粒提取物經電轉化大腸埃希菌J53,使轉化子大腸埃希菌J53對亞胺培南、美洛培南和阨他培南的MIC值增高2～64倍,PCR擴增轉化子大腸埃希菌J53 blaKPC基因均暘性.結論 本地區對碳青黴烯類抗生素耐藥的奇異變形桿菌,對阿米卡星的耐藥率相對較低.其碳青黴烯酶的基因型主要為位于質粒上的blaKPC-2基因型,易于在菌株間流行,臨床應引起高度重視.
목적 연구본지구근4년림상분리적비중복탄청매희류내약적기이변형간균가능존재적탄청매희류매기인형별.방법 회고분석2009도2012년림상분리기이변형간균적자료사선출비중복적탄청매희류항생소내약적균주15주；용농도제도법(E-시조법)검측기대아알배남(IPM)、미라배남(MEM)、액타배남(ETP)、배병사성(CIP)、아미잡성(AK)화미낙배소(MIN)적MIC치；Hodge시험진행산탄청매희매적학인,동시대시험균주진행내약기인적PCR확증검측급측서,대양성확증균주적질립제취물여첩담납내수적민감태대장애희균J53(E.coli J53)진행전전화,병대전전화후적균주진행목표기인적PCR확증급탄청매희류항생소MIC치적검측.결과 2009도2012년,사선도15주내탄청매희류항생소내약적균주；IPM、MEM、ETP적내약솔균위100％,CIP적내약솔위86.7％(13/15),AK적내약솔위33.3％ (5/15),MIN적내약솔위80％(12/15)；15주세균중Hodge시험양성7주,blaKPC기인양성11주,경측서검측학인위blaKPC-2형；11주blaKPC기인양성균주적질립제취물경전전화대장애희균J53,사전화자대장애희균J53대아알배남、미락배남화액타배남적MIC치증고2～64배,PCR확증전화자대장애희균J53 blaKPC기인균양성.결론 본지구대탄청매희류항생소내약적기이변형간균,대아미잡성적내약솔상대교저.기탄청매희매적기인형주요위위우질립상적blaKPC-2기인형,역우재균주간류행,림상응인기고도중시.
Objective To investigate the clinical distribution and the drug resistance of carbapenemresistant Proteus mirabilis strains isolated from 2009 to 2012 ; and to study carbapenemase genotypes of the isolates.Methods A total of 15 non-repetitive carbapenem-resistant Proteus mirabilis strains were selected from 422 Proteus mirabilis strains isolated from Ningbo First Hospital during January 2009 to December 2012.The minimal inhibitory concentrations (MIC) of 6 antibacterial agents,including imipenem (IPM),meropenem (MEM),ertapenem (ETP),ciprofloxacin (CIP),amikacin (AK) and minocycline (MIN),against 15 isolates were determined by E-test.The modified Hodge test (MHT) was used to detect the carbapenemase production in isolates.The PCR assay was performed to detect drug resistance genes of blaKPC,blaNDM-1,blaGES,blaSME,blaIMI-1/NmcA and blaSHV-38.Plasmids were extracted from the blaKPC-positive strains and then transformed into Escherichia coli J53 strains by electroporation.The transformed and untransformed Escherichia coli J53 strains were tested for MIC values and blaKPC gene by E-test and PCR respectively.Results The resistance rates of the 15 carbapenem-resistant Proteus mirabilis strains to IPM,MEM,ETP,CIP,AK and MIN were 100％,100％,100％,86.7％ (13/15),33.3％ (5/15) and 80％ (12/15),respectively.7 out of 15 strains were Hodge test positive,and 11 strains were blaKPC-2 positive.Results of PCR amplification showed that the transformed Escherichia coli J53 strains,whose MIC values to IPM,MEM,and ETP were increased by 2 to 64 times,were blaKPC-2 gene positive.Conclusion The carbapenem-resistant Proteus mirabilis strains in this study were resistant to many commonly used antibiotics,however,the resistance rates to AK were relatively low.The dominant carbapenemase genotype was blaKPC-2 carried by the plasmid.Attention should be paid to its easily transmissible feature among the strains in clinic.
