中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2014年
2期
136-139
,共4页
李莹莹%明亮%刘红春%康运凯%明昭
李瑩瑩%明亮%劉紅春%康運凱%明昭
리형형%명량%류홍춘%강운개%명소
糖尿病,2型%克雷伯菌,肺炎%抗药性,细菌%血清分型%毒力
糖尿病,2型%剋雷伯菌,肺炎%抗藥性,細菌%血清分型%毒力
당뇨병,2형%극뢰백균,폐염%항약성,세균%혈청분형%독력
Diabetes mellitus,type 2%Klebsiella pneumoniae%Drug resistance,bacterial%Serotyping%Virulence
目的 了解临床分离自2型糖尿病并发颌面部感染的98株肺炎克雷伯菌的脉冲场凝胶电泳(PFGE)分型和对常用抗菌药物的耐药情况,揭示其相关耐药机制,并对其毒力性进行探讨.方法 前瞻性研究.本研究采取定点连续抽样的方法,选择2010年3月至2012年10月郑州市7家医院颌面外科确诊为2型糖尿病合并颌面部感染患者为研究对象,共连续收集431例患者脓液标本,分离鉴定出98株肺炎克雷伯菌,K-B法检测肺炎克雷伯菌对15种抗菌药物的敏感性试验.用Xba I酶对菌株进行限制性酶切,PGFE分离酶切片段,Fingerprinting指纹图谱分析软件分析PFGE图谱,并分析PFGE基因组型与耐药表型之间的关系.产超广谱β内酰胺酶(ESBL)菌株检测应用双纸片协同及确认试验.通过PCR检测耐药基因型、血清型及毒力基因.并将TEM、SHV及CTX-M耐药基因的PCR扩增产物纯化、克隆后进行基因测序.黏液丝实验确定菌株的高黏液型(HM)表型.结果 98株肺炎克霄伯菌对15种抗生素的耐药情况严重,ESBL检出率达57.1％(56/98).PFGE将肺炎克雷伯菌分为13型,未发现优势带型及特异的ESBL电泳条带.PCR结果显示56株ESBL菌株中,产SHV型β内酰胺酶40株(占71.4％)、TEM型28株(占50.0％)和CTX-M型21株(占37.5％),测序结果证实均为TEM-1、CTX-M-3及多种SHV型.K1、K2、K3、K5、K20、K54和K57血清型及3种毒力基因均有检出,但不以强毒性血清型为主.HM阳性率为31.6％ (31/98).结论 2型糖尿病伴发颌面部感染来源的肺炎克雷伯菌的耐药性严重,耐药机制多样复杂,并且存在强毒性血清型和毒力基因,需引起临床的高度重视.
目的 瞭解臨床分離自2型糖尿病併髮頜麵部感染的98株肺炎剋雷伯菌的脈遲場凝膠電泳(PFGE)分型和對常用抗菌藥物的耐藥情況,揭示其相關耐藥機製,併對其毒力性進行探討.方法 前瞻性研究.本研究採取定點連續抽樣的方法,選擇2010年3月至2012年10月鄭州市7傢醫院頜麵外科確診為2型糖尿病閤併頜麵部感染患者為研究對象,共連續收集431例患者膿液標本,分離鑒定齣98株肺炎剋雷伯菌,K-B法檢測肺炎剋雷伯菌對15種抗菌藥物的敏感性試驗.用Xba I酶對菌株進行限製性酶切,PGFE分離酶切片段,Fingerprinting指紋圖譜分析軟件分析PFGE圖譜,併分析PFGE基因組型與耐藥錶型之間的關繫.產超廣譜β內酰胺酶(ESBL)菌株檢測應用雙紙片協同及確認試驗.通過PCR檢測耐藥基因型、血清型及毒力基因.併將TEM、SHV及CTX-M耐藥基因的PCR擴增產物純化、剋隆後進行基因測序.黏液絲實驗確定菌株的高黏液型(HM)錶型.結果 98株肺炎剋霄伯菌對15種抗生素的耐藥情況嚴重,ESBL檢齣率達57.1％(56/98).PFGE將肺炎剋雷伯菌分為13型,未髮現優勢帶型及特異的ESBL電泳條帶.PCR結果顯示56株ESBL菌株中,產SHV型β內酰胺酶40株(佔71.4％)、TEM型28株(佔50.0％)和CTX-M型21株(佔37.5％),測序結果證實均為TEM-1、CTX-M-3及多種SHV型.K1、K2、K3、K5、K20、K54和K57血清型及3種毒力基因均有檢齣,但不以彊毒性血清型為主.HM暘性率為31.6％ (31/98).結論 2型糖尿病伴髮頜麵部感染來源的肺炎剋雷伯菌的耐藥性嚴重,耐藥機製多樣複雜,併且存在彊毒性血清型和毒力基因,需引起臨床的高度重視.
목적 료해림상분리자2형당뇨병병발합면부감염적98주폐염극뢰백균적맥충장응효전영(PFGE)분형화대상용항균약물적내약정황,게시기상관내약궤제,병대기독력성진행탐토.방법 전첨성연구.본연구채취정점련속추양적방법,선택2010년3월지2012년10월정주시7가의원합면외과학진위2형당뇨병합병합면부감염환자위연구대상,공련속수집431례환자농액표본,분리감정출98주폐염극뢰백균,K-B법검측폐염극뢰백균대15충항균약물적민감성시험.용Xba I매대균주진행한제성매절,PGFE분리매절편단,Fingerprinting지문도보분석연건분석PFGE도보,병분석PFGE기인조형여내약표형지간적관계.산초엄보β내선알매(ESBL)균주검측응용쌍지편협동급학인시험.통과PCR검측내약기인형、혈청형급독력기인.병장TEM、SHV급CTX-M내약기인적PCR확증산물순화、극륭후진행기인측서.점액사실험학정균주적고점액형(HM)표형.결과 98주폐염극소백균대15충항생소적내약정황엄중,ESBL검출솔체57.1％(56/98).PFGE장폐염극뢰백균분위13형,미발현우세대형급특이적ESBL전영조대.PCR결과현시56주ESBL균주중,산SHV형β내선알매40주(점71.4％)、TEM형28주(점50.0％)화CTX-M형21주(점37.5％),측서결과증실균위TEM-1、CTX-M-3급다충SHV형.K1、K2、K3、K5、K20、K54화K57혈청형급3충독력기인균유검출,단불이강독성혈청형위주.HM양성솔위31.6％ (31/98).결론 2형당뇨병반발합면부감염래원적폐염극뢰백균적내약성엄중,내약궤제다양복잡,병차존재강독성혈청형화독력기인,수인기림상적고도중시.
Objective To investigate molecular typing and drug resistance patterns of 98 Klebsiella pneumoniae (K.pneumoniae) isolated from type 2 diabetes patients complicated with maxillofacial infection,to research the virulence and resistance mechanisms.Methods The study was a prospective study that adopted the method of continuous sampling from fixed location,from March 2010 to October 2012.The maxillofacial surgery patients diagnosed with type 2 diabetes complicated with maxillofacial infection were chosen in 7 hospitals in Zhengzhou as the research object,and a total of 431 pus sample were collected continuously,in which 98 strains K.pneumoniae were isolated and identified.The Kirby-Bauer disk diffusion test was conducted in 98 strains to determine the resistance to 19 antibacterial agents.K.pneumoniae chromosomal DNA were digested by restriction endonuclease Xba Ⅰ and analyzed by pulsed-field gel electrophoresis (PFGE).PFGE patterns of K.pneumoniae strains were analyzed using Fingerprinting software.The relationship between the molecular types and resistance phenotype was observed.The extended spectrum β-1actamase-producing K.pneumoniae were screened out by the double disc synergy test (DDST)Polymerase chain reaction (PCR) was used to detect resistant genotypes,serotype and virulence genes.The purified PCR products of resistant genes were cloned and sequenced.Hypermucoviscosity phenotype of all strains were determined by string test.Results Much severer drug-resistance for K.pneumoniae was identified and the result of extended-spectrum beta-lactamase producing rate was 57.1 ％.Ninety-eight strains were dispatched into 13 groups by PFGE.No dominant bands and specific extended-spectrum beta-lactamase DNA bands were found.The results of PCR showed that among the 56 strains of extended spectrum β-lactamase-producing K.pneumoniae,40 were positive for blaSHV (accounting for 71.4％),28 positive for blaTEM (accounting for 50.5％),21 positive for blaCTX-M (accounting tor 37.5％).The sequencing results were as follows:TEM-1,CTX-M-3 and a variety of SHV.Serotype K1,K2,K3,K5,K20,K54 and K57 and 3 kinds of virulence genes were detected,but not in strong toxicity-based.Hypermucoviscosity positive rate was 31.6％ (31/98).Conclusion Much severer drug resistance of K.pneumoniae in this study was identified and resistant mechanism was complex,in which strong toxicity serotype and virulence geues exist,which need more attention from clinical.
