用头孢西丁三相试验检测大肠埃希菌及肺炎克雷伯菌的质粒AmpC酶

目的：检测质粒介导的持续高产AmpC酶在大肠埃希菌及肺炎克雷伯菌中的携带率，为临床治疗提供指导。方法：采用头孢西丁纸片药敏试验（K-B法）作AmpC酶的初筛试验，头孢西丁三相试验间接法作AmpC酶的确证试验。结果：在检测的86株菌中，有9株菌K-B法初筛结果为阳性。在这9株菌中经头孢西丁三相试验确证有5株产AmpC酶，阳性率为5.8%。其中大肠埃希菌3株，肺炎克雷伯菌2株。结论：头孢西丁K-B法筛选试验结合头孢西丁三相试验可准确检出质粒介导的AmpC酶，可用于临床常规检测。     

AmpC酶在主要肠杆菌科和非发酵菌中的携带率及耐药性

目的：调查主要肠杆菌科细菌和非发酵菌产AmpC酶和ESBLs的状况及对常用抗菌药物的耐药性，指导临床合理用药。方法：常规培养分离细菌，应用VTTEK微生物自动分析仪和API鉴定系统鉴定病原菌；药敏试验采用K-B纸片扩散法；AmpC酶检测采用三维试验法；ESBLs检测采用双纸片确认试验。结果：主要肠杆菌科细菌产AmpC酶和ESBLs的阳性率分别为18．9％，52．8％，其中以大肠杆菌、肺炎克雷伯、产酸克雷伯、阴沟肠杆菌为主，同时产2种酶的菌株占15．4％；绿脓杆菌产AmpC酶和ESBLs酶的阳性率分别为18．2%，，42．0％，同时产2种酶的菌株占13．6％；不动杆菌的产酶率均在8．0％以下,产酶菌的耐药率高于非产酶菌，除绿脓杆菌外，产AmpC酶菌株的耐药率高于产ESBLs菌株．主要肠杆菌科细菌对泰能(IMP)的敏感性为99．5%，而对舒普深(CPZ)的耐药率平均为13．8％；非发酵菌对IMP，马斯平(FEP)、CPZ的耐药率平均分别为20．4％，44．3％，18．6％.结论：主要肠杆菌科细菌的产酶率高于非发酵菌，产AmpC酶的菌株已近20．0%，应引起临床的高度重视；绿脓杆菌产ES-BLs的菌株也较高，也应该加强监控．对产酶菌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；对重症感染患的治疗应根据药敏试验结果重点应用泰能或舒普深治疗。     

Emergence and wide dissemination of CTX-M-type ESBLs, and CMY-2- and DHA-1-type AmpC beta-lactamases in Korean respiratory isolates of Klebsiella pneumoniae

Respiratory isolates of Klebsiella pneumoniae in Korea during 2002-2003 were studied to determine the prevalence and types of extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated AmpC beta-lactamases (PABLs). ESBL-production was tested by double-disk synergy, and genotypes of beta-lactamases were determined by PCR and sequencing. ESBLs were detected in 28.4% of 373 isolates, and the most prevalent types were SHV-12 (63 isolates) and CTX-M-14 (9 isolates). Forty of 75 ESBL-producers (53.5%) also had PABLs: 21 isolates with CMY-2-like, 17 with DHA-1-like. Pulsed-field gel electrophoresis showed 19 types and 25 of 74 isolates had an identical pattern, indicating nosocomial spread. Dissemination of ESBL- and PABL-producing K. pneumoniae strains in Korea is a particular concern, as it limits the choice of antimicrobial agents for treatment of infections.     

产AmpC酶阴沟肠杆菌耐药的基础与临床

目的 对四川大学华西医院阴沟肠杆菌产AmpC酶菌株的检出率、耐药性及ampC结构基因序列进行分析．探讨阴沟肠杆菌产AmpC酶菌株的耐药性及临床特点。方法 采用琼脂稀释法对临床标本中分离的阴沟肠杆菌进行产AmpC酶株的筛选，用酶粗提物头孢西丁三维试验结合PCR法检测AmpC酶．并测定产AmpC酶菌株对9种抗菌药物的MIC值。对3株高度耐药阴沟肠杆菌产AmpC酶的结构基因和1株敏感菌进行PCR扩增并对产物序列进行分析，测序的菌株与E。cloacae p99进行核苷酸序列比较和推导的氨基酸序列比较。结果 阴沟肠杆菌产AmpC酶株的检出率为24．6％。产AmpC酶菌株多呈多重耐药，产AmpC酶耐药菌对9种抗菌药物的耐药率由高至低为头孢西丁、头孢噻肟、阿米卡星、氨曲南、头孢他啶、头孢哌酮／舒巴坦、环丙沙星、头孢吡肟、亚胺培南。阴沟肠杆菌耐药株ampC结构基因的核苷酸序列与E.cloacae p99的同源性为99％．推导的氨基酸序列仅Ala-58→Pro发生点突变。结论 四川大学华西医院阴沟肠杆菌产AmpC酶细菌检出率较高。产AmpC酶细菌多呈多重耐药，亚胺培南是治疗产AmpC酶细菌感染的较可靠的药物。阴沟肠杆菌产AmpC酶是其对β-内酰胺类抗生素耐药的原因之一，其ampC结构基因突变可能与其耐药性有关。     

Multidrug resistance mediated by co-carriage of extended-spectrum beta-lactamases,AmpC and New Delhi metallo-beta-lactamase-1 genes among carbapenem-resistant Enterobacteriaceae at five Indian medical centres

In this study, we evaluated the coexistence of extended-spectrum beta-lactamases (ESBL), AmpC and New Delhi metallo-beta-lactamase-1 (NDM-1) genes among carbapenem-resistant Enterobacteriaceae (CRE) recovered prospectively from patients at multiple sites. The study included 285 CRE strains from 2782 Gram-negative Bacilli collected from multiple centres during 2007–2010, of which 87 were characterised. Standard and reference laboratory methods were used for resistance determination. Detection of bla_NDM-1, bla_AmpC, bla_TEM, bla_SHV and bla_CTX-M was done by polymerase chain reaction. High levels of antimicrobial resistance observed among study isolates. Co-carriage of ESBLs, AmpC and NDM-1 was 26.3%. Nosocomial origin among the co-carriage isolates was 64.3%, with 9.2% associated mortality.     

High Prevalence of AmpC β-Lactamases in Clinical Isolates of Escherichia coli in Ilam,Iran

Objectives

Widespread use of β-lactam antibiotics could cause resistance to this group of antibiotics in pathogenic bacteria through the production of the enzyme β-lactamases. The aim of this study is to determine the molecular detection of AmpC β-lactamases among clinical Escherichia coli isolated from Ilam hospitals in Ilam, Iran.

Methods

One hundred and twelve clinical isolates of E. coli were collected from hospitalized patients and were identified by biochemical tests. They were evaluated for extended spectrum beta-lactamases (ESBLs) production, and the positive strains were subjected to AmpC enzymes; for detection of AmpC cluster genes, multiplex polymerase chain reaction was applied.

Results

The analysis showed 62.5% of isolates were ESBLs positive and that five strains revealed the AmpC cluster genes. This is the first report of FOXM cluster genes in E. coli in Iran.

Conclusion

Based on our results, the prevalence of AmpC β-lactamases is increasing in Iran, which caused failure in antibiotic therapy. So, the current study recommended the revision of antibiotic policy in Iranian hospitals.     

Molecular epidemiology,resistance profiles and clinical features in clinical plasmid-mediated AmpC-producing Enterobacteriaceae

During the 30 months of surveillance period, 85 pAmpC-producing isolates were detected (prevalence 0.56% overall): bla_CMY-2 gene in 70 E. coli, 2 K. pneumoniae and 6 P. mirabilis isolates; and the bla_DHA-1 gene in 4 E. coli and 3 K. pneumoniae. In 8.23% of them, other β-lactamases (predominantly OXA-1) were identified. All pAmpC-producing isolates were susceptible to carbapenems, whereas high resistance to nalidixic acid, ciprofloxacin and trimethoprim-sulfamethoxazole was observed among pAmpC-producing isolates (80%, 60%, and 44.7%, respectively). In hospital patients, predisposing factors such as prior antibiotic use, previous hospitalization, presence of an indwelling device, invasive urinary tract procedures and mechanical ventilation were observed. In the community setting, urinary tract infection was the most common type of infection related to pAmpC-producing isolates. A wide heterogeneity of clones was found among our E. coli isolates by PFGE, suggesting that this mechanism of resistance is not due to the dissemination of a clonal strain. Surveillance of these resistance mechanisms in the community is thus needed. Awareness of pAmpC dynamic is required to prevent introduction into hospitals and to control the spread of this emerging resistance within the community.     

Amp C酶检测方法的探讨

目的 建立简便检测肠杆菌科细菌中Amp C酶的方法。方法 分别采用双纸片确证法和多剂量协同法检测35株大肠埃希菌和肺炎克雷伯菌,并用三维试验和Amp C酶酶量直接测定法进行对比,同时检测这些菌产ESBL的情况。结果 双纸片确证法检测出12株Amp C酶,不受ESBL的干扰,与直接酶量检测法完全一致,敏感性高于三维试验,多刑量协同法敏感性较差。结论 双纸片确证法可作为临床检测Amp C酶的方法。     

107株阴沟肠杆菌产ESBLs和AmpC酶检测及耐药性分析

目的 了解近年来临床分离的阴沟肠杆菌对常用抗菌药物的敏感性,并检测分离株产超广谱β-内酰胺酶(ESBLs)和AmpC酶的情况,指导临床合理用药。 方法 收集2016年1-12月湖南中医药大学第一附属医院分离自临床标本的阴沟肠杆菌,采用Vitek-2 Compact进行常规药敏试验,采用表型确证试验检测ESBLs,采用三维试验检测AmpC酶。 结果 共收集非重复分离阴沟肠杆菌107株,主要来源于呼吸内科(26/107,24.3%)、骨伤科(21/107,19.6%)、中心ICU(20/107,18.7%)等科室,以痰液(38/107,35.5%)和伤口分泌物(29/107,27.1%)等标本为主;阴沟肠杆菌对青霉素类、三代头孢菌素和头霉素类高度耐药,对哌拉西林/他唑巴坦、头孢吡肟、亚胺培南耐药率分别为13.1%、22.4%、0.9%。共检测出单产ESBLs菌株30株(28.0%),单产AmpC酶的菌株35株(32.7%);同时产ESBLs和AmpC酶菌株14株(13.1%);同时产ESBLs和AmpC酶菌株对常用抗菌药物的耐药率高于不产酶株,差异均有统计学意义(均P＜0.05)。 结论 产ESBLs、AmpC酶是阴沟肠杆菌多重耐药的重要机制;阴沟肠杆菌产酶株的检测,对指导临床抗菌治疗具有重要意义。