鲍曼不动杆菌超广谱β-内酰胺酶检测及耐药性分析

为了解厦门中山医院鲍曼不动杆菌产超广谱β 内酰胺酶(ESBLs)情况及耐药现状,指导临床用药,采用美国临床实验室标准化委员会(NCCLS)推荐的纸片扩散法,测定 2002 ~2003年临床分离的 211株鲍曼不动杆菌产ESBLs的比例及其对头孢噻肟、阿米卡星等 13种抗菌药物耐药性特征。结果表明 211株鲍曼不动杆菌中ESBLs阳性 27株(占 12. 8% ),产ESBLs菌株对 13种抗菌药物的耐药率明显高于非产ESBLs株。鲍曼不动杆菌的产ESBLs株有较高的交叉耐药性,采用纸片确证实验检出ESBLs对于临床用药有一定的指导意义。     

鲍曼不动杆菌对碳青霉烯类抗生素的耐药性及其基因的分析

目的研究23株鲍曼不动杆菌对碳青霉烯类抗生素的耐药情况及对耐药基因分析,为临床用药提供依据。方法用珠海迪尔DL-96鉴定系统进行细菌鉴定及K-B法进行药敏试验,用碳青霉烯酶4种基因的特异性引物对其进行聚合酶链反应(PCR)扩增和基因型分析,并通过网上GenBank进行比对以确定编码酶基因的类型。结果 23株鲍曼不动杆菌对哌拉西林/他唑巴坦、左旋氧氟沙星、丁胺卡那霉素、多黏菌素B的耐药率分别为80%、45%、30%、10%,对其他抗生素的耐药率均在90%以上。携带D类碳青霉烯酶OXA-23基因有18株(78%),携带OXA-51基因有15株(65%),OXA-24、OXA-58基因引物PCR扩增为阴性,随机各抽取3株OXA-23基因阳性株进行测序后通过在网上GenBank比对与OXA-23标准株99%同源,OXA-51基因阳性株与OXA-51标准株98%同源。结论耐碳青霉烯类抗生素的鲍曼不动杆菌对多黏菌素的耐药率最低,其次是丁胺卡那霉素,其中以携带OXA-23型碳青霉烯酶基因为主,应引起临床高度关注,防止在院内广泛传播。     

Profiles of phenotype resistance to antibiotic other than β-lactams in Klebsiella pneumoniae ESBLs-producers, carrying blaSHV genes

Extended spectrum β-lactamases production is one of the most common mechanism of resistance to extended spectrum β-lactam antibiotics is increasing worldwide. Twenty five strains of Klebsiella pneumoniae isolated from clinical specimens were tested. Based on the phenotypic confirmatory test all these strains were defined as ESBL producers named ESBL(+). The plasmid DNA from each strains was used to investigate the presence of blaSHV genes responsible for extended spectrum β-lactamases production. Moreover, susceptibility of these strains to antibiotic other than β-lactams in was tested.     

重症监护室耐亚胺培南鲍曼不动杆菌的耐药性分析

目的探讨温州医学院附属第一医院重症监护室（ICU）耐亚胺培南鲍曼不动杆菌的耐药特性。方法对2006年1月至2007年12月ICU分离的菌株采用常规方法进行菌株分离,经革兰染色、氧化酶试验等初筛,再经VITEK-32型全自动微生物分析仪进行菌株鉴定和药敏试验。结果共检测到98株鲍曼不动杆菌,其中耐亚胺培南的鲍曼不动杆菌为50株（占51.0%）,标本来源大部分为痰液（占94.9%）。耐亚胺培南鲍曼不动杆菌对常用16种抗菌药物耐药率〉50%的有15种,耐药率〉90%有3种,分别为氨苄西林（98.0%）、头孢唑林（99.0%）和呋喃妥因（99.0%）,对抗菌药物耐药率最低仅为头孢哌酮/舒巴坦（10.2%）,另外检测到对所用抗菌素泛耐为4株（8.0%）;而对亚胺培南敏感的鲍曼不动杆菌对常用16种抗菌药物耐药率〉50%的仅为4种,耐药率〉90%同样为氨苄西林（100%）、头孢唑林（97.9%）和呋喃妥因（97.9%）,耐药率比较低的为美罗培南（10.4%）、妥布霉素（16.7%）和环丙沙星（16.7%）,耐药率最低也为头孢哌酮/舒巴坦（6.3%）。结论 ICU分离耐亚胺培南鲍曼不动杆菌多重耐药性严重,出现了泛耐菌株;临床可根据药敏试验选用抗菌药物,一般条件下耐亚胺培南鲍曼不动杆菌建议选用头孢哌酮／舒巴坦。     

694株鲍曼不动杆菌的耐药性分析

目的了解大连市友谊医院临床分离的鲍曼不动杆菌的分布及耐药状况,为临床治疗鲍曼不动杆菌感染提供参考。方法对该院2007年至2009年住院患者送检的标本中分离到的694株鲍曼不动杆菌的分布及药敏结果做回顾性分析。结果鲍曼不动杆菌2007年检出112株（7．1％）,2008年检出210（11．3％）株,2009年检出372（14．7％）株;在所有临床送检的标本中,痰标本中的检出率最高,占88．7％;在对13种抗生素的药敏试验中,对美罗培南最为敏感,其次对头孢哌酮／舒巴坦较为敏感。结论鲍曼不动杆菌的检出率逐年增高,耐药率也逐年增高。     

多重耐药鲍曼不动杆菌的耐药性及其耐药基因分析

目的:探讨多重耐药鲍曼不动杆菌(MDR-Ab)的耐药性及其耐药基因,为临床合理选择抗菌药物提供依据。方法:回顾性分析2018年1月至2018年12月鲍曼不动杆菌感染的住院患者信息。使用VITEK-32微生物分析仪/梅里埃药敏卡片GN13鉴定MDR-Ab 95株。采用聚合酶链式反应(多重PCR)检测MDR-Ab携带相关耐药基因。结果:95株MDR-Ab对头孢类抗菌药物耐药率为100%。对氨苄西林-舒巴坦和头孢哌酮-舒巴坦耐药率分别为95.79%和81.05%,对美罗培南和亚胺培南耐药率分别为56.84%和57.89%,对庆大霉素和阿米卡星耐药率均为88.42%,对环丙沙星和左氧氟沙星耐药率分别为100%和88.42%,对四环素、米诺环素、替加环素耐药率分别为87.37%、16.84%和9.47%,对多粘菌素B耐药率为1.05%。95株MDR-Ab中携带β-内酰胺酶中A类酶耐药基因TEM、PER分别95株和25株,D类酶耐药基因OXA-51、carO和adeB各95株,OXA-23基因90株。携带消毒剂耐药基因qacE 60株。携带16S r RNA甲基化酶耐药基因armA 75株。每株MDR-Ab除携带TEM+carO+adeB+OXA-51四种基因外,另同时携带四种基因20株(21.05%),三种基因38株(40.00%)。结论:MDR-Ab对多种抗菌药物的耐药率较高,携带的耐药基因型主要为TEM、carO、adeB及OXA-51。携带多种耐药基因是MDR-Ab耐药重要原因。加强医院感染防控、合理应用抗菌药物对于延缓泛鲍曼不动杆菌耐药性发展具有重要的临床意义。     

产β-内酰胺酶鲍曼不动杆菌基因型研究

目的了解临床科室分离的鲍曼不动杆菌耐药状况并分析多重耐药株产β-内酰胺酶基因型,为有效的临床治疗和医院感染控制提供实验室依据。方法采用纸片扩散（K-B）法测定临床分离的312株鲍曼不动杆菌对13种抗菌药物的敏感性,对其中120株多重耐药株用聚合酶链反应（PCR）扩增β-内酰胺酶基因,并对其扩增产物进行基因测序。结果鲍曼不动杆菌对美洛培南、亚胺培南、头孢哌酮/舒巴坦和替卡西林/克拉维酸的耐药率分别为0.0%、1.0%、30.8%和31.4%,其余抗菌药物耐药率在38.5%-80.1%;120株多重耐药菌株中,β-内酰胺酶基因分布AmpC型基因阳性率为66.7%（80/120）、SHV-12型基因阳性率为14.2%（17/120）,PER-1型基因阳性率为16.7%（20/120）,CTX-M-9型基因阳性率为8.2%（10/120）,TEM-1型基因阳性率为9.2%（11/120）;VER-1,CTX-M-1,CTX-M-2,OXA型均阴性。同时携带2种基因型有16株,携带3种基因型有14株。结论临床分离的鲍曼不动杆菌流行株主要是产AmpC酶,且呈多重耐药。     

The role of bla(OXA-like carbapenemase) and their insertion sequences (ISS) in the induction of resistance against carbapenem antibiotics among Acinetobacter baumannii isolates in Tehran hospitals

This study aimed to evaluate the occurrence and dissemination of bla(OXA-like) carbapenemase genes and their insertion sequences among Acinetobacter baumannii isolates, taken from different hospitals in Tehran city and also their roles in the induction of resistance to carbapenem drugs. A total number of 100 non duplicate Acinetobacter baumannii with different origins, were isolated from patients with proved nosocomial infections at eight university hospital in Tehran city. Antimicrobial susceptibility of these strains was done by E-test against 7 antimicrobial agents according to CLSI guideline. PCR of bla(OXA-51-like), bla(OXA-23-like), bla(OXA-24-like), bla(OXA-58-like), IS(ABA-1), IS(1133) was carried out by specialized primers and then these strains were typed by REP-fingerprinting. Colistin, imipenem and meropenem were the most sensitive antibiotics against Acinetobacter baumannii isolates with 96%, 51% and 51% sensitivity respectively. All the isolates had a bla(OXA-51-like) intrinsic to these species. The rates of bla(OXA-23), 23 and 58-like were 38%, 32% and 1% respectively. Coexistence of bla(OXA-51/23/24-like) was observed among 16% of these isolates. All bla(OXA-23-like) carbapenemase genes had only one IS(ABA1). REP fingerprinting showed 5 genotypes among carbapenem resistant isolates, 16 of them being genotype A. This study emphasized on the major role of bla(OXA-like) carbapenemase, particularly bla(OXA-23-like) carbapenemase and their IS(ABA1), in the dissemination of carbapenem resistant Acinetobacter baumannii. This study confirmed a presumptive role of IS element neighboring the carbapenemase gene in the elevation of resistance to carbapenem drug among Acinetobacter baumannii isolates for the first time in Iran.     

Antibmicrobial susceptibility of Serratia marcescens isolated from hospitalized patients in 2003 - 2005

500 strains of Serratia marcescens isolated in 2003-2005 were examined for drug susceptibility. By using several phenotypic methods it was shown that 67.6% of these strains produced ESBLs. Strains ESBL(-) and ESBL(+) were compared, paying special attention to their susceptibility to various antibiotics. It was revealed that strains ESBL(+) were much more resistant to majority of the investigated drugs. The biggest differences were in the case of amikacin and gentamicin, sensitive about 50% of ESBL(-) and 10% of ESBL(+), ciprofloxacin, sensitive 42% of ESBL(-) and 6.3% of ESBL(+) and trimethoprim/ sulphametoxazole, sensitive 45.8% of ESBL(-) and 9.4% of ESBL(+). Strains ESBL(-) retained a high susceptibility to ceftazidime (68.9%) and cefepime (71%). All strains ESBL(-) as well as ESBL(+) were susceptible to imipenem and meropenem. 78.9% of ESBL(-) and 67.3% of investigated ESBL(+) were susceptible to piperacillin/ tazobactam.     

346株鲍曼不动杆菌临床分布及耐药率分析

目的了解鲍曼不动杆菌引起医院感染的特点以及对抗菌药物的耐药性的变化趋势。方法应用美国BD公司Phoenix^TM100全自动细菌和药敏系统鉴定仪对所分离的364株鲍曼不动杆菌进行鉴定和药敏试验,并进行统计学分析。结果 346株鲍曼不动杆菌来自痰液、伤口分泌物、中段尿、静脉血及大便,分别占58.96%、30.92%、5.49%、4.34%、0.29%。药敏结果显示鲍曼不动杆菌对多粘菌素和米诺环素敏感性最高,耐药率低于10.4%。结论加强鲍曼不动杆菌的耐药监测,了解其耐药性变迁,可合理指导用药,有效控制鲍曼不动杆菌耐药菌株的产生。     

鲍曼不动杆菌耐药程度与其主动外排泵蛋白的相关性研究

目的:探讨鲍曼不动杆菌耐药程度与其主动外排泵蛋白的相关性。方法:首先用纸片扩散法检测64株临床鲍曼不动杆菌对8种抗菌药物的敏感性;将其分为A组(0～2种抗生素耐药)、B组(对3～5种抗生素耐药)和C组(对6～8种抗生素耐药);检测64株临床鲍曼不动杆菌对罗丹明6G的外排情况,筛选出罗丹明6G外排明显增加的菌株;并用逆转录-聚合酶链反应(RT-PCR)方法检测主动外排泵基因AdeABC的表达水平。结果:64株鲍曼不动杆菌中有4株对0～2种抗生素耐药(A组),对3～5种抗生素耐药的有33株(B组),对6～8种抗生素耐药的有27株(C组);多重耐药组鲍曼不动杆菌罗丹6G外排明显增高,外排程度A组相似文献   

Genetic basis of antibiotic resistance in pathogenic Acinetobacter species

Antibiotic resistance in Acinetobacter spp., particularly Acinetobacter baumannii, is increasing rapidly. A. baumannii possesses two intrinsic β-lactamase genes, in addition to weak permeability and efflux systems, that together confer a natural reduced susceptibility to antibiotics. In addition, numerous acquired mechanisms of resistance have been identified in A. baumannii. The very high genetic plasticity of A. baumannii allows an accumulation of resistance determinants that give rise to multidrug resistance at an alarming rate. The role of novel genetic elements, such as resistance islands, in concentrating antibiotic resistance genes in A. baumannii requires detailed investigation in the near future.     

In-silico modeling of a novel OXA-51 from β-lactam-resistant Acinetobacter baumannii and its interaction with various antibiotics

Acinetobacter baumannii, one of the major Gram negative bacteria, causes nosocomial infections such as pneumonia, urinary tract infection, meningitis, etc. β-lactam-based antibiotics like penicillin are used conventionally to treat infections of A. baumannii; however, they are becoming progressively less effective as the bacterium produces diverse types of β-lactamases to inactivate the antibiotics. We have recently identified a novel β-lactamase, OXA-51 from clinical strains of A. baumannii from our hospital. In the present study, we generated the structure of OXA-51 using MODELLER9v7 and studied the interaction of OXA-51 with a number of β-lactams (penicillin, oxacillin, ceftazidime, aztreonam and imipenem) using two independent programs: GLIDE and GOLD. Based on the results of different binding parameters and number of hydrogen bonds, interaction of OXA-51 was found to be maximum with ceftazidime and lowest with imipenem. Further, molecular dynamics simulation results also support this fact. The lowest binding affinity of imipenem to OXA-51 indicates clearly that it is not efficiently cleaved by OXA-51, thus explaining its high potency against resistant A. baumannii. This finding is supported by experimental results from minimum inhibitory concentration analysis and transmission electron microscopy. It can be concluded that carbapenems (imipenem) are presently effective β-lactam antibiotics against resistant strains of A. baumannii harbouring OXA-51. The results presented here could be useful in designing more effective derivatives of carbapenem.     

高龄患者血液感染常见革兰阴性杆菌及其耐药性分析

目的分析本院80~100岁高龄患者血液感染常见革兰阴性杆菌的种类及其耐药状况,为本院合理使用抗生素提供依据。方法采用BacT/Alert 3D血培养仪对血液标本进行阳性鉴定;VITEK-2Compact全自动微生物鉴定仪进行鉴定;K-B纸片扩散法对抗菌药物进行敏感性测定;使用WHONET 5.4分析软件分析数据。结果本院高龄患者血液培养阳性标本中共分离出革兰阴性杆菌108株,以肠杆菌科细菌为主,其次为非发酵菌,前者主要为大肠埃希菌52株(48.15%)和肺炎克雷伯菌37株(34.26%),后者主要包括铜绿假单胞菌10株(9.26%)和鲍曼不动杆菌8株(7.41%)。其中大肠埃希菌和肺炎克雷伯菌对亚胺培南的耐药率分别为1.92%和13.51%,两者对氨苄西林、氨苄西林/舒巴坦、头孢唑林、头孢呋辛的耐药率均高于50.00%;铜绿假单胞菌和鲍曼不动杆菌对亚胺培南的耐药率分别为20.00%和25.00%,后者对其他抗菌药物的耐药率均高于前者。结论碳青酶烯类抗生素可作为本院高龄患者常见革兰阴性杆菌所致血液感染的首选药物;但在治疗中应考虑细菌的耐药特点及患者的代谢特点合理选择抗生素。     

社区与医院感染中肺炎克雷伯杆菌和大肠埃希菌耐药性分析

目的探讨社区和医院感染中肺炎克雷伯杆菌和大肠埃希菌产ESBLs的情况及耐药特性。方法采用体外扩散确证试验检测ESBLs,同时用Micro scan wat RA way-40系统全自动细菌鉴定/药敏分析仪及K-B琼脂扩散法进行细菌鉴定和体外药敏试验。结果社区感染标本中分离出肺炎克雷伯杆菌79株,产ESBLs20株,阳性率为25.3%,大肠埃希菌177株,产ESBLs27株,阳性率为15.3%;医院感染标本中分离出肺炎克雷伯杆菌82株,产ES-BLs33株,阳性率为40.2%,大肠埃希菌135株,产ESBLs42株,阳性率为31.1%,社区与医院感染菌株产ESBLs比较差异均有统计学意义(P均<0.05);ESBLs阳性菌株对多种抗生素耐药,其耐药性明显高于ESBLs阴性菌株。结论肺炎克雷伯杆菌和大肠埃希菌产ESBLs菌株在临床分离率较高,医院感染标本要显著高于社区感染标本,并且对多种抗生素具有高度耐药性,产ESBLs菌株耐药性显著高于不产ESBLs菌株,临床上应加强对ESBLs的控制,以防感染流行。     

鲍曼不动杆菌对碳青霉烯类抗生素的耐药性分析

目的了解鲍曼不动杆菌的耐药情况,并检测耐碳青霉烯类鲍曼不动杆菌的耐药基因,为指导临床合理用药、控制院内感染提供依据。方法利用K-B法检测45株鲍曼不动杆菌临床分离株的耐药情况,通过改良Hodge试验、Carba NP试验和EDTA协同试验对多重耐药鲍曼不动杆菌的碳青霉烯酶进行表型检测,并采用PCR技术检测鲍曼不动杆菌携带OXA-23和NDM-1型耐药基因的情况。结果 45株鲍曼不动杆菌临床分离株中共筛出42株多重耐药菌株;利用改良Hodge试验和Carba NP试验检出36株碳青霉烯酶阳性菌株;采用PCR扩增出OXA-23,未扩增出NDM-1。结论鲍曼不动杆菌耐药情况严重,且耐药基因OXA-23携带率高,治疗时应根据药敏试验结果合理用药。     

不同时期肠杆菌科细菌产ESBLS及耐药性的监测

目的对不同时期肠杆菌科细菌产超广谱β-内酰胺酶（ESBLS）及耐药性进行监测。方法采用生物梅里埃ATB药敏试验板测定肠杆菌科细菌中大肠埃希菌和肺炎克雷伯菌对抗生素的敏感性,同时用表型确证法检测ESBLS。结果肠杆菌科细菌2006-2008年与2009-2011年两个时期相比,ESBLS的检出率增长近50%。对亚胺配南、头孢吡肟、头孢西丁耐药率较低,对哌拉西林/他唑巴坦、阿米卡星的耐药率次之,对其他β-内酰胺类、氨基糖苷类、喹诺酮类及磺胺类抗生素的耐药率较高,且两个时期相比耐药率增加明显,差异具有统计学意义（P〈0.05）。结论临床产ESBLS菌株不断增多,多重耐药现象越来越严重,临床实验室应当把ESBLS的检测作为常规监测工作,有效指导临床选用抗菌药物。     

鲍曼不动杆菌耐药性及碳青霉烯酶检测

目的了解临床分离鲍曼不动杆菌对临床常用抗菌药物的耐药性及其产碳青霉烯酶情况,为临床抗感染治疗提供依据。方法采用MicroScan WalkAway-40全自动微生物分析仪对240株临床分离的鲍曼不动杆菌进行鉴定和药敏试验,改良的Hodge试验检测耐碳青霉烯类鲍曼不动杆菌产生的碳青霉烯酶,并采用SPSS 13.0软件进行统计分析。结果 240株鲍曼不动杆菌对亚胺培南的耐药率最低(20.8%),对青霉素类、头孢菌素类、喹诺酮类、复方新诺明的耐药率均大于50%;耐碳青霉烯类鲍曼不动杆菌碳青霉烯酶的检出率为64.0%。结论临床分离鲍曼不动杆菌耐药情况严重,碳青霉烯酶是鲍曼不动杆菌对碳青霉烯类抗菌药物耐药的主要原因。     

1939株革兰阴性杆菌的耐药监测分析

目的 了解广州华侨医院2007年至2009年分离的1 939株革兰阴性杆菌的耐药状况,为临床合理使用抗生素提供依据.方法 利用梅里埃VITEK-2 Compact微生物分析仪鉴定细菌,K-B法进行药敏试验,采用WHONET 5.4 软件统计分析药敏结果.结果 2007年至2009年分离出的革兰阴性杆菌1 939株,居前...     

鲍曼不动杆菌耐药性监测与碳青霉烯酶基因型研究

目的：对鲍曼不动杆菌耐药性及碳青霉烯酶基因型进行研究,以指导临床合理应用抗生素。方法：收集青岛市海慈医疗集团2009年6月至2010年6月从临床分离的鲍曼不动杆菌60株,用琼脂稀释法测定最低抑菌浓度（M IC）,改良Hodge试验检测碳青霉烯酶,用PCR法检测OXA-23,OXA-24,OXA-58基因,并对PCR产物进行测序。结果：①鲍曼不动杆菌检出率前两位是ICU病房和呼吸科病房,分别占32.3%和27.4%,多重耐药鲍曼不杆菌阳性率最高的是ICU,为70.6%（12/17）,其次为呼吸科病房,为35.0%（7/20）,哌拉西林、哌拉西林/他唑巴坦、头孢曲松、头孢他啶、头孢吡肟、亚胺培南、美罗培南、庆大霉索、阿米卡星、环丙沙星、左氧氟沙星、加替沙星、头孢哌酮/舒巴坦、氨曲南耐药率分别为92.3%、55.4%、88.6%、86.3%、80.3%、30.0%、35.0%、76.6%、79.6%、75.1%、87.1%、48.3%、42.0%和79.6%.②在21株耐碳青霉烯类鲍曼不动杆菌,有14株碳青霉烯酶表型阳性,检出率为66.7%,有18株PCR扩增出OXA-23基因,检出率85.7%,全部菌株blaOXA-24及blaOXA-58PCR扩增均为阴性,PCR产物测序表明与鲍曼不动杆菌（AY795964.1）blaOXA-23基因序列100%同源。结论：鲍曼不动杆菌多重耐药性严重;表型和基因型检测证实本院临床分离鲍曼不动杆菌对碳青霉烯类耐药机制主要是产OXA-23型酶。