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
2014年
6期
423-430
,共8页
吴伟元%陆坚%卢月梅%吴劲松%李文青%程锦娥%梁训宏%吴文苑%刘映霞
吳偉元%陸堅%盧月梅%吳勁鬆%李文青%程錦娥%樑訓宏%吳文苑%劉映霞
오위원%륙견%로월매%오경송%리문청%정금아%량훈굉%오문원%류영하
奇异变形杆菌%超广谱β-内酰胺酶%AmpC酶%qnrD%脉冲场凝胶电泳
奇異變形桿菌%超廣譜β-內酰胺酶%AmpC酶%qnrD%脈遲場凝膠電泳
기이변형간균%초엄보β-내선알매%AmpC매%qnrD%맥충장응효전영
Proteus mirabilis%Extended-spectrum β-lactamase%AmpC enzyme%qnrD%Pulsed-field gel electrophoresis
目的：调查深圳市人民医院产超广谱β-内酰胺酶（ ESBL ）和/或AmpC酶奇异变形杆菌的流行及其分子特征。方法使用ESBL表型初筛和确证试验以及AmpC酶纸片法检测产ESBL和/或AmpC酶奇异变形杆菌。 PCR扩增和DNA测序确定ESBL、AmpC酶基因型及其上游插入序列、质粒介导喹诺酮类耐药（ PMQR）基因型和染色体gyrA、gyrB和parC基因的喹诺酮类耐药决定区（ QRDRs）突变位点，以及整合酶基因和1类整合子基因盒。脉冲场凝胶电泳（ PFGE）分析菌株的同源性。结果2004-2010年我院临床共分离130株奇异变形杆菌，13株（10％）产 ESBL，3株（2．3％）产AmpC酶；产ESBL菌从0％～9．1％（2004-2009年）迅速上升至29．4％（2010年）。最常见ESBL基因型为 blaCTX-M-14（n＝7），其他 ESBL 基因型包括 blaCTX-M-65（n＝3）、blaCTX-M-55（n＝1）、blaCTX-M-24（n＝1）和blaPER-1（n＝1），系国内首次临床分离出携带blaPER-1奇异变形杆菌；2株产AmpC酶菌基因型为blaCMY-2，1株产AmpC酶菌基因型未知。91．7％（11/12）奇异变形杆菌的blaCTX-M上游和1株blaCMY-2上游均携带ISEcp1；1株blaPER-1上游携带ISPa12。66．7％（10/15）产ESBL和/或AmpC酶奇异变形杆菌携带qnrD（n＝5）和/或aac-Ib-cr（n＝8）。12株环丙沙星耐药产ESBL和/或AmpC酶菌在QRDRs中均携带GyrA上1个点突变（S83I）和ParC上1个点突变（S80I或S80R），其中6株还同时携带GyrB上1个点突变（E466D）。86．7％（13/15）产ESBL和/或AmpC酶菌携带1类整合子。15株产ESBL和/或AmpC酶奇异变形杆菌共获14种不同PFGE型别。结论产CTX-M型酶是我院奇异变形杆菌对超广谱头孢菌素耐药的主要机制，产ESBL和/或AmpC酶奇异变形杆菌大多携带qnrD和/或aac-Ib-cr基因。
目的：調查深圳市人民醫院產超廣譜β-內酰胺酶（ ESBL ）和/或AmpC酶奇異變形桿菌的流行及其分子特徵。方法使用ESBL錶型初篩和確證試驗以及AmpC酶紙片法檢測產ESBL和/或AmpC酶奇異變形桿菌。 PCR擴增和DNA測序確定ESBL、AmpC酶基因型及其上遊插入序列、質粒介導喹諾酮類耐藥（ PMQR）基因型和染色體gyrA、gyrB和parC基因的喹諾酮類耐藥決定區（ QRDRs）突變位點，以及整閤酶基因和1類整閤子基因盒。脈遲場凝膠電泳（ PFGE）分析菌株的同源性。結果2004-2010年我院臨床共分離130株奇異變形桿菌，13株（10％）產 ESBL，3株（2．3％）產AmpC酶；產ESBL菌從0％～9．1％（2004-2009年）迅速上升至29．4％（2010年）。最常見ESBL基因型為 blaCTX-M-14（n＝7），其他 ESBL 基因型包括 blaCTX-M-65（n＝3）、blaCTX-M-55（n＝1）、blaCTX-M-24（n＝1）和blaPER-1（n＝1），繫國內首次臨床分離齣攜帶blaPER-1奇異變形桿菌；2株產AmpC酶菌基因型為blaCMY-2，1株產AmpC酶菌基因型未知。91．7％（11/12）奇異變形桿菌的blaCTX-M上遊和1株blaCMY-2上遊均攜帶ISEcp1；1株blaPER-1上遊攜帶ISPa12。66．7％（10/15）產ESBL和/或AmpC酶奇異變形桿菌攜帶qnrD（n＝5）和/或aac-Ib-cr（n＝8）。12株環丙沙星耐藥產ESBL和/或AmpC酶菌在QRDRs中均攜帶GyrA上1箇點突變（S83I）和ParC上1箇點突變（S80I或S80R），其中6株還同時攜帶GyrB上1箇點突變（E466D）。86．7％（13/15）產ESBL和/或AmpC酶菌攜帶1類整閤子。15株產ESBL和/或AmpC酶奇異變形桿菌共穫14種不同PFGE型彆。結論產CTX-M型酶是我院奇異變形桿菌對超廣譜頭孢菌素耐藥的主要機製，產ESBL和/或AmpC酶奇異變形桿菌大多攜帶qnrD和/或aac-Ib-cr基因。
목적：조사심수시인민의원산초엄보β-내선알매（ ESBL ）화/혹AmpC매기이변형간균적류행급기분자특정。방법사용ESBL표형초사화학증시험이급AmpC매지편법검측산ESBL화/혹AmpC매기이변형간균。 PCR확증화DNA측서학정ESBL、AmpC매기인형급기상유삽입서렬、질립개도규낙동류내약（ PMQR）기인형화염색체gyrA、gyrB화parC기인적규낙동류내약결정구（ QRDRs）돌변위점，이급정합매기인화1류정합자기인합。맥충장응효전영（ PFGE）분석균주적동원성。결과2004-2010년아원림상공분리130주기이변형간균，13주（10％）산 ESBL，3주（2．3％）산AmpC매；산ESBL균종0％～9．1％（2004-2009년）신속상승지29．4％（2010년）。최상견ESBL기인형위 blaCTX-M-14（n＝7），기타 ESBL 기인형포괄 blaCTX-M-65（n＝3）、blaCTX-M-55（n＝1）、blaCTX-M-24（n＝1）화blaPER-1（n＝1），계국내수차림상분리출휴대blaPER-1기이변형간균；2주산AmpC매균기인형위blaCMY-2，1주산AmpC매균기인형미지。91．7％（11/12）기이변형간균적blaCTX-M상유화1주blaCMY-2상유균휴대ISEcp1；1주blaPER-1상유휴대ISPa12。66．7％（10/15）산ESBL화/혹AmpC매기이변형간균휴대qnrD（n＝5）화/혹aac-Ib-cr（n＝8）。12주배병사성내약산ESBL화/혹AmpC매균재QRDRs중균휴대GyrA상1개점돌변（S83I）화ParC상1개점돌변（S80I혹S80R），기중6주환동시휴대GyrB상1개점돌변（E466D）。86．7％（13/15）산ESBL화/혹AmpC매균휴대1류정합자。15주산ESBL화/혹AmpC매기이변형간균공획14충불동PFGE형별。결론산CTX-M형매시아원기이변형간균대초엄보두포균소내약적주요궤제，산ESBL화/혹AmpC매기이변형간균대다휴대qnrD화/혹aac-Ib-cr기인。
Objective To investigate the prevalence and molecular characteristics of the extended -spectrum β-lactamase ( ESBL) and AmpC enzyme-producing Proteus mirabilis ( P.mirabilis) strains isola-ted in Shenzhen People′s Hospital.Methods The production of ESBLs and AmpC enzymes by P.mirabilis isolates were detected by a screening and confirmatory test for ESBLs and AmpC disk test , respectively .The PCR assays followed by DNA sequencing of the products were employed to analyze the multiple genes inclu -ding the ESBLs genes, AmpC genes, insertion sequences (ISs) upstream of the ESBLs or AmpC genes, plasmid -mediated quinolone resistance ( PMQR ) determinants , quinolone resistance-determining region (QRDR) genes , the integrase genes, and class1 integron cassette.The epidemiological analysis of the iso-lates was performed by pulsed field gel electrophoresis .Results There were 130 P.mirabilis clinical iso-lates collected from Shenzhen People′s Hospital in China during the year 2004 to 2010.Among them, 13 isolates (10%) produced ESBLs, that accounted for 0%-9.1%in the year 2004-2009 and up to 29.4%in 2010, and 3 isolates (2.3%) produced AmpC enzymes.The predominant genotype of ESBLs -producing isolateswas b al CTX-M-14(n=7), followed by blaCTX-M-65(n=3), blaCTX-M-55(n=1), blaCTX-M-24(n=1) and blaPER-1 (n =1).The clinical isolate of PER-1-producing P.mirabilis was reported for the first time in China.Twoisolates carried an AmpC β-lactamase gene of blaCMY-2 and one isolate carried an unidentified AmpC gene .ISEcp1 located upstream of blaCTX-M and blaCMY-2 were detected in 91.7% (11/12) of CTX-M-producing isolatesand one CMY-2-producing isolate, respectively.ISPa12 was present upstream of blaPER-1 in one studiedisolate.Approximately 66.7% (10/15) of ESBL and /or AmpC-producing isolates harbored PMQR genes including2 carrying qnrD, 5 carrying aac-Ib-cr and 3 carrying both qnrD and aac-Ib-cr.Twelve ESBL and /orAmpC-producers with high level of resistance to ciprofloxacin carried the similar mutation profiles of S 83I inGyrA, S80I or S80R in ParC and among them, six strains showed E466D mutation in GyrB.Approximately86.7% (13/15) of ESBL and/or AmpC-producing isolates carried class 1 integron.Fourteen PFGE typeswere observed among 15 ESBL and/or AmpC-producers.Conclusion The prevalence of CTX-M β-lactamasesin P.mirabilis isolates contributed to the increased resistance to extended -spectrum cephalosporins.The qnrD and/or aac-Ib-cr genes were detected among the most of ESBL and /or AmpC-producing P.mirabilis clinical isolates.
