铜绿假单胞菌和阴沟肠杆菌超超广谱β-内酰胺酶的检测

目的：了解多重耐药的铜绿假单胞菌和阴沟肠杆菌的主要耐药机制超超广谱β-内酰胺酶即超广谱β-内酰胺酶(ESBLs)和高产AmpC酶的产生情况。方法：建立一种简易快速地检测超超广谱β-内酰胺酶的K-B药敏试验法：首先选择5种药敏纸片：IMP、FOX、FEP、CTX、CD03,依据纸片的抑菌环直径综合判断分析。怀疑产AmpC酶的菌株,用OBs抑制试验进一步证实。结果：使用该法分别检测产AmpC酶、ESBLs的4株标准株和多重耐药的70株铜绿假单胞菌和30株阴沟肠杆菌,结果各种标准株检测无误。70株铜绿假单胞菌中产AmpC酶的有40株,产ESBLs的有18株,同时产AmpC酶和ESBLs的是5株,还有9株细菌两类酶检测均阴性,原因待分析。同时利用此试验法检测铜绿假单胞菌的诱导阳性率为42／70(60％)和阴沟肠杆菌的诱导阳性率为16／30(53．3％)。结论：此法简易快速,能较好地鉴别产ESBLs或和AmpC酶株,并适用于临床常规检验。     

肠杆菌诱导型AmpC酶对第三代和第四代头孢菌素的影响

目的探讨诱导型AmpC酶对三代、四代头孢菌素的影响及其治疗策略。方法采用药敏纸片扩散法,用FOX药敏纸片作为诱导剂,以CAZ、CTX和FEP作为指示剂,检测肠杆菌的诱导AmpC酶的产生。结果待测的196株菌株中有101株为产诱导型AmpC酶阳性（51．5％）,其中阴沟肠杆菌58株（67．4％）,产气肠杆菌18株（52．9％）,沙雷菌属共8株（40％）,费劳地枸椽酸杆菌6株（42．8％）,聚团肠杆菌6株（33．3％）,奇异变形杆菌5株（20．8％）,其余95株产诱导型AmpC酶为阴性。产诱导型AmpC酶阳性菌株中无一株对FEP纸片的抑菌圈直径有影响,诱导型AmpC酶试验均为阴性。结论对于鉴定确认为诱导型AmpC酶阳性菌株的感染,即使实验报告三代头孢菌素为敏感,也应慎用或避免使用第三代头孢菌素的治疗,以第四代头孢菌素如头孢吡肟为首选药。     

革兰阴性杆菌ESBLs和AmpC酶的检测及耐药分析

目的检测引起医院感染的革兰阴性杆菌携带产ESBLs和去阻遏AmpC酶状况,探讨各菌对临床常用抗生素的主要耐药机制及耐药性,为临床制定合理使用抗生素策略提供依据。方法采用全自动微生物分析仪（VITEK-32）做细菌鉴定和药敏试验,用纸片扩散确证法检测超广谱β-内酰胺酶（ESBLs）,用三维法检测高水平表达染色体编码的AmpC酶。结果158株革兰阴性杆菌ESBLs检出率为26．6％,主要菌为大肠埃希菌（45．2％）、肺炎克雷伯菌（42．9％）、阴沟肠杆菌（11．9％）。AmpC酶检出率为10．1％,主要为鲍曼不动杆菌（43．8％）、阴沟肠杆菌（25％）;上述产酶细菌均对青霉素和一、二、三代头孢菌素、磺胺类、喹诺酮类、氨基糖苷类耐药,对亚胺培南敏感。结论革兰阴性杆菌耐药机制主要是产超广谱β-内酰胺酶和AmpC酶,这些产酶菌株均出现多重耐药。     

阴沟肠杆菌产AmpC酶菌和非产酶菌的耐药性研究

目的：探讨阴沟肠杆菌分布特征及产AmpC酶菌和非AmpC酶菌的耐药性。方法：对临床分离的158株阴沟肠杆菌分布科室、感染部位及对16种抗生素耐药性进行分析,并通过酶粗提物头孢西丁三维试验结合PCR法检测AmpC酶。结果：标本来源主要为患者的痰液、尿液、创口分泌物等,科室以重症监护室为多,感染部位以呼吸道为主,耐药性较高的抗生素为头孢西丁、头孢噻肟、头孢曲松等,158株阴沟肠杆菌中产AmpC酶菌株共33株,产AmpC酶阳性率占总菌株数20．9％,产AmpC酶菌株对各种抗生素的耐药率比不产AmpC酶的明显增高。结论：阴沟肠杆菌的耐药与产AmpC酶有关,治疗首选亚胺培南。     

血液透析患者感染的菌群分布和耐药性分析

目的了解引起血液透析感染菌群分布及耐药情况。方法用K-B法进行药敏试验,对革兰阴性菌采用ESBLs确认试验检测ESBLs,头孢西丁三维试验检测AmpC。结果本组分离的259株细菌中,革兰阴性菌179株,占69.1%,主要致病菌为大肠埃希菌、肺炎克雷伯菌、阴沟肠杆菌。革兰阳性菌80株,占30.9%,以金黄色葡萄球菌、凝固酶阴性葡萄球菌居前2位。革兰阴性杆菌ESBLs、AmpC酶总检出率分别为42.4%、39.1%,单产ESBLs、单产AmpC、同产ESBLs 高产AmpC酶、ESBLs 诱导AmpC酶菌株依次占15.6%、12.3%、16.8%、10.1%。革兰阳性分离株除对万古霉素、替考拉宁、喹奴普汀-达福普汀、利福平的耐药率较低外,其余抗生素的耐药率均大于50.0%。革兰阴性分离株对亚胺培南、美罗培南、头孢哌酮/他唑巴坦的耐药率分别为5.02%、3.35%和6.70%,产酶株较非产酶株具有较高的耐药率。结论血液透析患者细菌感染以金黄色葡萄球菌、凝固酶阴性葡萄球菌、大肠埃希菌、肺炎克雷伯菌、阴沟肠杆菌为主,万古霉素、替考拉宁对革兰阳性菌,亚胺培南、美罗培南和头孢哌酮/他唑巴坦对革兰阴性菌具有良好的抗菌活性。     

产AmpC酶阴沟肠杆菌的基因分析及其耐药性

探讨昆明地区阴沟肠杆菌的耐药性及与结构基因ampC和调节基因ampD的相关性。通过K-B法检测阴沟肠杆菌的药敏情况,头孢西丁三维试验检测AmpC酶,PCR法扩增ampC和ampD基因。结果显示74株阴沟肠杆菌经头孢西丁三维试验检测,产AmpC酶的有17株,检出率为22.3%,而且产酶菌株抗生素敏感率低于非产酶菌株。ampC基因扩增阳性率为89.2%(66/74);64株ampD基因阳性率为86.5%(64/74)。实验证实昆明地区产酶阴沟肠杆菌耐药状况严重,与结构基因ampC和调节基因ampD密切相关。     

多重耐药阴沟肠杆菌质粒型AmpC酶DHA-1基因的检出

目的测定56株同时耐头孢噻肟、庆大霉素和环丙沙星的阴沟肠杆菌质粒型AmpC酶基因型。方法先后用头孢西丁纸片法、三维试验、等电聚焦及酶抑制试验和微量稀释法进行表型检测。接合试验证实酶基因的转移性。多重聚合酶链反应以及基因测序等方法鉴定质粒型AmpC酶基因型。结果受试的56株细菌中有5株三维试验阳性,其中1株能转移接合,接合子多重聚合酶链反应扩增结果呈阳性,等电点为7．8,基因测序表明和DHA-1型AmpC酶一致。结论我国的多重耐药阴沟肠杆菌已获得质粒型DHA基因,DHA基因可通过转化、接合等方式转移给其他细菌且易于传播,因此应加强监控以防DHA基因在革兰阴性菌中流行。     

肺炎克雷伯菌超广谱β-内酰胺酶和AmpC酶基因检测及耐药性分析

目的 了解下呼吸道感染肺炎克雷伯菌超广谱β-内酰胺酶（ESBLs）和质粒AmpC酶的产生情况及耐药性,研究AmpC酶的基因型别.方法 用纸片扩散确证法检测ESBLs;用酶提取三维试验检测AmpC酶,聚合酶链反应（PCR）扩增AmpC酶的基因,DNA序列测定检测AmpC酶的基因型;K-B法检测细菌耐药性.结果 58株下呼吸道感染肺炎克雷伯菌中ESBLs阳性21株,AmpC酶阳性5株,其中3株ESBLs和AmpC酶均阳性,5株AmpC酶阳性菌中,4株扩增出DHA基因,经测序均为DHA-1,1株扩增出MIR基因.产酶菌株的耐药性明显高于非产酶株.结论 肺炎克雷伯菌中ESBL＊s和AmpC酶均有较高的检出率,AmpC酶以DHA基因型为主.产ESBLs和AmpC酶是肺炎克雷伯菌耐药的主要原因.     

铜绿假单胞菌ESBLs和AmpC酶的检测及耐药性分析

目的了解铜绿假单胞菌临床分离株ESBLs和AmpC酶的产生及对常用抗菌药物敏感性,指导临床合理选用抗生素。方法常规培养分离细菌,采用VITEK-60型全自动细菌鉴定仪鉴定细菌;按NCCLS推荐的双纸片确证法和K-B纸片扩散法检测ESBLs和药敏试验;采用头孢西丁纸片扩散法筛选疑产AmpC酶阳性菌株,确诊采用三维试验。结果铜绿假单胞菌产ESBLs和AmpC酶总检出率分别为40.8%和38.2%,其中,单产AmpC酶、单产ESBLs和同产AmpC酶+ESBLs检出率分别为19.7%、26.3%和14.5%。药敏试验显示:产酶株的耐药性明显高于非产酶株,耐药现象在同产AmpC酶和ESBLs菌株中更为严重。结论台州地区临床分离的铜绿假单胞菌产ESBLs和AmpC酶菌株检出率较高。产AmpC酶和ESBLs的菌株呈高度耐药,临床上对产酶菌株引起感染的治疗应根据细菌药敏试验结果,合理选择有效的抗菌药物联合治疗,减少产酶菌株的产生和流行。     

产妇会阴切口感染肺炎克雷伯菌质粒介导AmpC酶基因型检测与分析

摘要：目的 回顾性分析我院产妇会阴切口感染分离的肺炎克雷伯菌质粒介导AmpC酶基因型。方法 采用K-B纸片扩散法对临床分离的肺炎克雷伯菌进行AmpC酶初筛;头孢西丁三维试验确证产AmpC酶;采用UNIQ-10柱式质粒小量抽提试剂盒提取细菌质粒;采用聚合酶链反应（PCR）和DNA测序检测分析质粒AmpC酶基因型别。结果 三维试验检出15株产AmpC酶菌株,经PCR扩增15株菌株均在405 bp处出现阳性条带,经DNA测序证实为DHA-1型质粒AmpC酶。结论 我院存在质粒介导AmpC酶流行菌株,质粒AmpC酶主要以DHA-1型为主。     

Molecular characteristics of extended-spectrum beta-lactamases and qnr determinants in Enterobacter species from Japan

The incidence of extended-spectrum β-lactamases (ESBLs) has been increasing worldwide, but screening criteria for detection of ESBLs are not standardized for AmpC-producing Enterobacteriaceae such as Enterobacter species. In this study, we investigated the prevalence of ESBLs and/or AmpC β-lactamases in Japanese clinical isolates of Enterobacter spp. and the association of plasmid-mediated quinolone resistance (PMQR) determinants with ESBL producers. A total of 364 clinical isolates of Enterobacter spp. collected throughout Japan between November 2009 and January 2010 were studied. ESBL-producing strains were assessed by the CLSI confirmatory test and the boronic acid disk test. PCR and sequencing were performed to detect CTX-M, TEM, and SHV type ESBLs and PMQR determinants. For ESBL-producing Enterobacter spp., pulsed-field gel electrophoresis (PFGE) was performed using XbaI restriction enzyme. Of the 364 isolates, 22 (6.0%) were ESBL producers. Seven isolates of Enterobacter cloacae produced CTX-M-3, followed by two isolates producing SHV-12. Two isolates of Enterobacter aerogenes produced CTX-M-2. Of the 22 ESBL producers, 21 had the AmpC enzyme, and six met the criteria for ESBL production in the boronic acid test. We found a significant association of qnrS with CTX-M-3-producing E. cloacae. The 11 ESBL-producing Enterobacter spp. possessing bla(CTX-M), bla(SHV), or bla(TEM) were divided into six unique PFGE types. This is the first report about the prevalence of qnr determinants among ESBL-producing Enterobacter spp. from Japan. Our results suggest that ESBL-producing Enterobacter spp. with qnr determinants are spreading in Japan.     

Evaluation of a commercial beta-glucuronidase test for the rapid and economical identification of Escherichia coli

A commercial beta-glucuronidase (beta-GUR) test for the rapid and economical identification of Escherichia coli was evaluated. A total of 762 clinical strains and 228 environmental isolates were studied. More than 95% of the E. coli strains were found to be beta-GUR positive. Thirty-one clinical isolates of Shigella sonnei, 10 of Enterobacter cloacae, eight of Enterobacter aerogenes, nine of Citrobacter freundii and one of Salmonella enteritidis also gave positive results. The enzyme beta-GUR was also detected in two environmental strains of E. cloacae and one C. freundii. A comparative study between the beta-GUR test and the conventional identification system was carried out in 233 consecutive isolates of lactose positive enterobacteria. Agreement was observed in 223 cases and 190 E. coli strains were correctly identified using this test. Discrepancies were found in 10 cases: nine E. coli were beta-GUR negative and one C. freundii was beta-GUR positive. Escherichia coli was the only species positive for both beta-GUR and indole tests. This procedure permits a rapid, easy, precise and inexpensive identification of E. coli. beta-GUR positive Enterobacter strains have not previously been described.     

Evaluation of a commercial β-glucuronidase test for the rapid and economical identification of Escherichia coli

A commercial β-glucuronidase (β-GUR) test for the rapid and economical identification of Escherichia coli was evaluated. A total of 762 clinical strains and 228 environmental isolates were studied. More than 95% of the E. coli strains were found to be β-GUR positive. Thirty-one clinical isolates of Shigella sonnei , 10 of Enterobacter cloacae , eight of Enterobacter aerogenes, nine of Citrobacter freundii and one of Salmonella enteritidis also gave positive results. The enzyme β-GUR was also detected in two environmental strains of E. cloacae and one C. freundii. A comparative study between the β-GUR test and the conventional identification system was carried out in 233 consecutive isolates of lactose positive enterobacteria. Agreement was observed in 223 cases and 190 E. coli strains were correctly identified using this test. Discrepancies were found in 10 cases: nine E. coli were β-GUR negative and one C. freundii was β-GUR positive. Escherichia coli was the only species positive for both β-GUR and indole tests. This procedure permits a rapid, easy, precise and inexpensive identification of E. coli. β-GUR positive Enterobacter strains have not previously been described.     

产ESBLs肺炎克雷伯菌AmpC酶的检测及耐药性分析

目的了解深圳市人民医院产ESBLs肺炎克雷伯菌中产AmpC酶的情况及其耐药性。方法收集产ESBLs肺炎克雷伯菌临床株126株,应用Tris-EDTA纸片法检测AmpC酶。用琼脂稀释法测定菌株对11种抗生素的最低抑菌浓度（MIC）。结果126株ESBLs阳性的肺炎克雷伯菌中12株检出AmpC酶,检出率为9.5%。AmpC阳性菌株对头孢西丁、头孢他啶、氨曲南、氨苄西林/舒巴坦和阿莫西林/克拉维酸的耐药率达100%,对阿米卡星和哌拉西林/他唑巴坦的耐药率分别为83.3%和33.3%,其中头孢西丁、头孢他啶、氨曲南、阿莫西林/克拉维酸、阿米卡星和哌拉西林/他唑巴坦的耐药率显著高于AmpC阴性株（P〈0.05）。结论深圳市人民医院产ESBLs肺炎克雷伯菌中检出AmpC酶阳性株,其耐药性强于单产ESBLs菌株。     

500例疑似需氧菌性阴道炎患者阴道分泌物镜检和生化检测结果对比分析

目的探讨需氧菌性阴道炎（AV）生化检测法的临床应用价值。方法使用镜检和生化检测对500例疑似AV患者阴道分泌物进行检查,对检测结果进行对比分析。结果在500例疑似AV患者中,镜检法检出阳性患者409例,生化检测法共检出阳性患者414例。两种方法的诊断符合率为96.60%,二者呈中度一致性（Kappa=0.783,P〈0.05）。以镜检法为标准,生化检测法的敏感度为98.53%、特异性为87.91%。结论生化检测法对AV诊断具有很高的敏感度和特异度,且与镜检法符合率高,值得临床推广使用。