天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2015年
2期
166-169,170
,共5页
曹阳%刘双庆%魏殿军%陈薇
曹暘%劉雙慶%魏殿軍%陳薇
조양%류쌍경%위전군%진미
大肠杆菌%交叉感染%毒力因子类%多态性,单核苷酸%fimH基因
大腸桿菌%交扠感染%毒力因子類%多態性,單覈苷痠%fimH基因
대장간균%교차감염%독력인자류%다태성,단핵감산%fimH기인
Escherichia coli%cross infection%virulence factors%polymorphism,single nucleotide%fimH gene
目的：了解老年患者医院感染肠外致病性大肠埃希菌（ExPEC）30种毒力因子的检出情况及其分子流行病学特点。方法收集天津地区引起老年患者医院感染的非重复肠外致病性大肠埃希菌140株，采用多重PCR方法检测其毒力因子基因的存在情况，比较不同标本来源ExPEC毒力因子基因的检出率。从检出fimH基因的菌株中选取50株，进行fimH基因的扩增与测序，将全部测序结果与大肠埃希菌对照株CFT037、UTI89和参考株K-12 Gen?Bank数据库中的fimH基因序列均输入软件DNAMAN 6.0.3.93中寻找单核苷酸多态性（SNPs），后将53个fimH基因序列输入软件MEGA4，进行基因分型和树状图的构建。分型结果结合病例回顾，综合分析医院内大肠埃希菌克隆流行情况。结果 ExPEC中毒力因子基因fimH、traT、fyuA、iutA和kpsMT II的检出率较高，非尿标本来源ExPEC的毒力因子基因kpsM II、K5、papC、papEF、papG allele II (Internal)、papA、cnf1(CNF)、sfa/focDE和rfc的检出率均比尿标本来源ExPEC的高。53株菌fimH基因SNPs分析显示，在57个多态位点中发现有60种SNPs。53株菌被分为25个基因型，其中有11组菌株的基因指纹图谱分别完全一致。未发现不同医院的菌株具有相同的基因型。结论老年患者医院感染的肠外致病性大肠埃希菌携带多种毒力因子基因；非尿标本来源的大肠埃希菌可能比尿液标本来源的菌株致病性更强；fimH基因SNPs分型适用于医院大肠埃希菌克隆流行的调查，值得在今后的临床工作中推广。
目的：瞭解老年患者醫院感染腸外緻病性大腸埃希菌（ExPEC）30種毒力因子的檢齣情況及其分子流行病學特點。方法收集天津地區引起老年患者醫院感染的非重複腸外緻病性大腸埃希菌140株，採用多重PCR方法檢測其毒力因子基因的存在情況，比較不同標本來源ExPEC毒力因子基因的檢齣率。從檢齣fimH基因的菌株中選取50株，進行fimH基因的擴增與測序，將全部測序結果與大腸埃希菌對照株CFT037、UTI89和參攷株K-12 Gen?Bank數據庫中的fimH基因序列均輸入軟件DNAMAN 6.0.3.93中尋找單覈苷痠多態性（SNPs），後將53箇fimH基因序列輸入軟件MEGA4，進行基因分型和樹狀圖的構建。分型結果結閤病例迴顧，綜閤分析醫院內大腸埃希菌剋隆流行情況。結果 ExPEC中毒力因子基因fimH、traT、fyuA、iutA和kpsMT II的檢齣率較高，非尿標本來源ExPEC的毒力因子基因kpsM II、K5、papC、papEF、papG allele II (Internal)、papA、cnf1(CNF)、sfa/focDE和rfc的檢齣率均比尿標本來源ExPEC的高。53株菌fimH基因SNPs分析顯示，在57箇多態位點中髮現有60種SNPs。53株菌被分為25箇基因型，其中有11組菌株的基因指紋圖譜分彆完全一緻。未髮現不同醫院的菌株具有相同的基因型。結論老年患者醫院感染的腸外緻病性大腸埃希菌攜帶多種毒力因子基因；非尿標本來源的大腸埃希菌可能比尿液標本來源的菌株緻病性更彊；fimH基因SNPs分型適用于醫院大腸埃希菌剋隆流行的調查，值得在今後的臨床工作中推廣。
목적：료해노년환자의원감염장외치병성대장애희균（ExPEC）30충독력인자적검출정황급기분자류행병학특점。방법수집천진지구인기노년환자의원감염적비중복장외치병성대장애희균140주，채용다중PCR방법검측기독력인자기인적존재정황，비교불동표본래원ExPEC독력인자기인적검출솔。종검출fimH기인적균주중선취50주，진행fimH기인적확증여측서，장전부측서결과여대장애희균대조주CFT037、UTI89화삼고주K-12 Gen?Bank수거고중적fimH기인서렬균수입연건DNAMAN 6.0.3.93중심조단핵감산다태성（SNPs），후장53개fimH기인서렬수입연건MEGA4，진행기인분형화수상도적구건。분형결과결합병례회고，종합분석의원내대장애희균극륭류행정황。결과 ExPEC중독력인자기인fimH、traT、fyuA、iutA화kpsMT II적검출솔교고，비뇨표본래원ExPEC적독력인자기인kpsM II、K5、papC、papEF、papG allele II (Internal)、papA、cnf1(CNF)、sfa/focDE화rfc적검출솔균비뇨표본래원ExPEC적고。53주균fimH기인SNPs분석현시，재57개다태위점중발현유60충SNPs。53주균피분위25개기인형，기중유11조균주적기인지문도보분별완전일치。미발현불동의원적균주구유상동적기인형。결론노년환자의원감염적장외치병성대장애희균휴대다충독력인자기인；비뇨표본래원적대장애희균가능비뇨액표본래원적균주치병성경강；fimH기인SNPs분형괄용우의원대장애희균극륭류행적조사，치득재금후적림상공작중추엄。
Objective To examine the detection rate of 30 known virulence factors (VFs) of extraintestinal pathogenic Escherichia coli(ExPEC), and to investigates the epidemiology of ExPEC in elderly nosocomial infection. Methods A to?tal of 140 ExPEC clinical isolates from elderly nosocomial patients in hospitals in Tianjin were investigated. Multiplex PCR was performed to detect the 30 virulence factors among the E.coli strains and the detection rate of virulence factors for Ex?PEC were compared between isolates from different sites of infection.Fifty E. coli strains were shown to carry fimH gene that was amplified and sequenced. These sequences were used besides3 references strains (CFT037、UTI89 and K-12 ) to detect SNPs of fimH gene using DNAMAN Version 6.0.3.93 these 53 fimH sequences were used for genotyping and building dendrogram by MEGA4 software. Results In ExPEC, the following virulence factor genes, fimH, traT, fyuA, iutA and kpsMT II, had a higher detection rate than those of the rest . The following virulence factor genes, kpsMT II, K5, papC, pa?pEF ,papG allele II (Internal), papA, cnf1 (CNF), sfa/focDE and rfc had a a higher detectionrate from non-urine origin sam?ples than those from urine origin samples. fimH SNPs analysis of the 50 clinical isolated samples and 3 references samples showed 60 SNPs at 57 polymorphic sites. The fimH SNPs analysis classified the 53 strains into 25 genotype. The genetic fin?gerprintings of 11 isolates were exactly the same. Conclusion Many kinds of virulence factors can be found in ExPEC of el?derly nosocomial infection. The ExPEC strain isolated from non-urine origin had a stronger pathogenicity than those from urine-origin specimens. fimH SNPs analysis is suitable for molecular epidemiological investigation of ExPEC in hospital.
