耐亚胺培南鲍曼不动杆菌医院内感染流行的分子机制研究

目的研究耐亚胺培南鲍曼不动杆菌的耐药谱特征及其医院内感染流行和耐药性产生的分子机制,为临床防治提供依据.方法4株耐亚胺培南鲍曼不动杆菌分离自2002年10月至2003年1月外科重症监护病房的感染患者,采用纸片扩散法及E-test进行药物敏感性检测及MIC值测定,肠杆菌科基因组内重复一致序列聚集合酶链反应(ERIC-PCR)进行克隆株的DNA分型,耐药质粒转移及消除试验、等电聚焦电泳、PCR扩增β-内酰胺酶基因及其克隆测序以识别其耐药基因和进行质粒定位.结果4株菌除对头孢哌酮/舒巴坦复合制剂的MIC值较低外,对头孢菌素类、氨基糖甙类和氟喹喏酮类等抗生素均显示出了较高水平的多重耐药性;DNA分型证实为同一克隆株;产OXA-23型碳青霉烯酶和PER-1型超广谱β-内酰胺酶(ESBLs);OXA-23定位在质粒上,PER-1定位在染色体上.结论本组耐亚胺培南鲍曼不动杆菌为多重耐药株,同一克隆株在不同感染个体间的相互传播导致了本次医院内感染的流行,产OXA-23和PER-1型β-内酰胺酶是其耐药性产生的重要原因.     

PCR genotyping of Acinetobacter hospital isolates

A total of 13 Acinetobacter baumannii-13TU isolates obtained from patients of the Perm regional clinical hospital during the period of 1 year, were genotyped in the polymerase chain reaction (PCR) with universal primers. Acinetobacter cultures could not be distinguished by phenotypic tests and antibiogram and were resistant to B-lactams and gentamicin. According to the results of amplification the obtained isolates could be subdivided into two groups (with 6 and 7 respectively). The detection of "epidemic" strains, including those capable of prolonged (for more than 5 months) persistence in hospital environment, may be indicative of the growing role of Acinetobacter as the causative agent in nosocomial infection.     

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。     

下呼吸道不动杆菌医院感染的危险因素及耐药性分析

目的了解引起下呼吸道医院感染的不动杆菌耐药现状及存在的危险因素。方法用常规的方法对下呼吸道的标本进行病原菌的培养及分离,用全自动微生物分析仪VITEK 2对细菌进行鉴定及药物敏感性试验,同时对由不动杆菌引发的158例院内感染患者的危险因素进行分析。结果不动杆菌引起下呼吸道医院感染相关的危险因素主要为使用糖皮质激素类药物或机械插管或患有糖尿病等;除亚胺培南、左氧氟沙星、头孢吡肟、头孢哌酮、头孢哌酮/舒巴坦及头孢他啶等对不动杆菌有较好的体外抗菌活性外(耐药率小于40.0%),临床常用的其他多种抗菌药物耐药较严重,耐药率均在40.0%以上。结论糖皮质激素,机械插管,糖尿病等是不动杆菌引起下呼吸道医院感染的主要危险因素,不动杆菌对临床常用的抗生素有多重耐药性。     

Acinetobacter infection--an emerging threat to human health

The genus Acinetobacter comprises a complex and heterogeneous group of bacteria, many of which are capable of causing a range of opportunistic, often catheter-related, infections in humans. However, Acinetobacter baumannii, as well as its close relatives belonging to genomic species 3 ("Acinetobacter pittii") and 13TU ("Acinetobacter nosocomialis"), are important nosocomial pathogens, often associated with epidemic outbreaks of infection, that are only rarely found outside of a clinical setting. These organisms are frequently pandrug-resistant and are capable of causing substantial morbidity and mortality in patients with severe underlying disease, both in the hospital and in the community. Several epidemic clonal lineages of A. baumannii have disseminated worldwide and seem to have a selective advantage over non-epidemic strains. The reasons for the success of these epidemic lineages remain to be elucidated, but could be related to the potential of these organisms to achieve very dynamic reorganization and rapid evolution of their genome, including the acquisition and expression of additional antibiotic resistance determinants, under fluctuating environmental and selective conditions.     

某医院2007年至2009年医院感染不动杆菌属耐药调查分析

目的了解医院感染不动杆菌属细菌对临床常用抗菌药物耐药现状。方法对2007年至2009年852例不动杆菌属医院感染病例进行回顾性调查及耐药情况统计。结果不动杆菌属主要来源于痰液(60%)和血液(8%)标本,3年间17种抗生素耐药水平差异均有统计学意义;852株不动杆菌属对17种抗菌药物总耐药率:头孢哌酮/舒巴坦35.13%、亚胺培南/西司他丁46.4%、美罗培南51.0%、阿米卡星64.7%、四环素66.7%、左氧氟沙星67.1%、替卡西林/克拉维酸67.8%、环丙沙星68.0%、头孢吡肟68.7%、哌拉西林/他唑巴坦70.1%、头孢他啶70.5%、头孢曲松71.6%、头孢噻肟72.8%、庆大霉素73.5%、哌拉西林73.8%、复方新诺明77.3%和头孢哌酮86.8%。结论不动杆菌属细菌对临床常用抗菌药物耐药现象严重,仅头孢哌酮/舒巴坦、亚胺培南/西司他丁、美罗培南保持相对高的抗菌活性。     

Etiology of nosocomial surgical infections caused by gram-negative bacteria, and profile of their antibiotic resistance

Microbiological monitoring of microbial landscape of gram-negative bacteria--etiological agents of nosocomial surgical infections--was performed as well as their level of antibiotic resistance was studied. Two hundred forty-four strains were isolated. Antimicrobial susceptibility was assessed by the method of serial microdilutions. Spectrum of Gram-negative microorganisms was represented by 3 groups: fermenting (62.7%), nonfermenting (34.8%), and nonidentified microbes (2.5%). Spectrum of gram-negative etiological agents of nosocomial surgical infections was represented mainly by Pseudomonas aeruginosa, Acinetobacter baumanii, Escherichia coli, and Klebsiella. Most active antibacterial drugs against studied strains were carbapenems (imipenem and meropenem).     

Distribution of blaOXA genes among carbapenem-resistant Acinetobacter baumannii nosocomial strains in Poland

Acinetobacter baumannii is an important nosocomial pathogen occurring particularly in intensive care (ICU) as well as burn therapy units (BTU). A. baumannii strains have emerged as resistant to almost all antimicrobial agents, including carbapenems. b-lactamase-mediated resistance is the most common mechanism for carbapenem resistance in this species. Carbapenem-hydrolysing class D b-lactamases - OXA are widespread among A. baumannii strains. It is suggested that ISAba1 plays an important role in drug resistance. The aims of the study were detection of OXA encoding genes and presence of ISAba1. The study included the total of 104 isolates of carbapenem-resistant A. baumannii, obtained from patients hospitalized in ICU and BTU of Specialized Hospital in Krakow. Multiplex PCR was applied for detection of selected OXA carbapenemases encoding genes. PCR analysis showed the presence of bla OXA-51-like gene and ISAba1 in all isolates. 46 strains carried bla OXA-51-like and bla OXA-23-like genes while 48 bla OXA-51-like and bla OXA-40-like genes. 3 isolates carried: bla OXA-51-like , bla OXA-23-like and bla OXA-40-like genes. 7 strains encoded an OXA-51-like carbapenemase but were negative for enzymes belonging to the other families tested. Comparative analysis of ICU and BTU isolates revealed the dominance of: bla OXA-51-like and bla OXA-40-like among ICU while bla OXA-51-like and bla OXA-23-like in BTU.     

医院感染不动杆菌和肠球菌的分离及耐药性调查分析

目的了解医院感染不动杆菌和肠球菌的分离及耐药情况,为临床合理用药提供依据。方法采用回顾性资料,对医院2008年9月至2010年12月,从临床送检标本中分离到的不动杆菌和肠球菌及其药敏结果进行统计分析。结果从临床标本中分离到191株不动杆菌和319株肠球菌,不动杆菌主要来源是痰液,达77.50%,肠球菌主要来源是尿液、痰液,分别为31.70%和28.8%。不动杆菌对亚胺培南耐药率最低,为10.3%,对头孢噻肟耐药率最高,为7 2.0%,而其余各种抗生素耐药率均处于47%～60%,肠球菌对万古霉素耐药率最低,为2.0%,耐药率最高是四环素为63.9%,对高浓度的庆大霉素和链霉素耐药率分别是52.8%和39.4%。结论医院感染不动杆菌和肠球菌的分离及耐药性高,且呈多重耐药,临床要关注。     

Integron-bearing methicillin-resistant coagulase-negative staphylococci in South China, 2001–2004

A total of 53 methicillin-resistant coagulase-negative staphylococci strains isolated in a hospital in Guangzhou, China, were analyzed to detect class 1 integrons and SCC mec typing. Thirty strains had the class 1 integrase ( intI1 ) gene and 26 strains possessed the 3' conserved region of qacE Δ 1 - sul1 . Four different types of gene cassette arrays were found and a highly prevalent array of dfrA12-orfF-aadA2 gene cassettes was observed. Thirty class 1 integron-positive coagulase-negative staphylococci strains were subjected to Southern hybridization analysis; the result showed that class 1 integrons were located on chromosome, not plasmid. According to the results of SCC mec typing for 30 integron-bearing MRCNS strains, five, 15 and five strains belonged to type I, II and III SCC mec , respectively, and five strains were untypeable. For 23 non-integron-bearing methicillin-resistant coagulase-negative staphylococci strains, four, nine and seven strains belonged to type I, II and III SCC mec , respectively, and three strains were untypeable. None of the strains belonged to type IV or V. Twenty-three coagulase-negative staphylococci isolates of three Staphylococcal species that contained the dfrA12-orfF-aadA2 gene cassette array were phylogenetically unrelated to each other by randomly amplified polymorphic DNA, indicating that the gene cassettes might be disseminated in the clinical strains by a horizontal gene transfer.     

Class 1 integrons in Pseudomonas aeruginosa isolates from clinical settings in Amazon region, Brazil

A hundred and six Pseudomonas aeruginosa isolates from clinical cases were screened using PCR for the presence of integrons and associated resistance gene cassettes. Forty-four isolates harboured class 1 integrons (41.5%), of which 29 isolates (66%) also carried gene cassettes. The aacA gene was most frequently found within class 1 integrons (69%), followed by blaOXA family genes (52%). From class 1 integron-positive strains, we detected a total of 15 isolates (34%) carrying no gene cassettes. Restriction fragment-length polymorphism analysis of the integrons variable region revealed some identical structures, as well as distinct profiles indicating heterogeneity among these cassette regions. Multiresistance was observed in 71% of isolates, nevertheless no strong correlation was observed between integron presence and multiresistance. This is the first report showing class 1 integron prevalence and gene cassette content in P. aeruginosa isolates from clinical settings in the Brazilian Amazon.     

Integron-bearing methicillin-resistant coagulase-negative staphylococci in South China, 2001-2004.

A total of 53 methicillin-resistant coagulase-negative staphylococci strains isolated in a hospital in Guangzhou, China, were analyzed to detect class 1 integrons and SCCmec typing. Thirty strains had the class 1 integrase (intI1) gene and 26 strains possessed the 3' conserved region of qacEDelta1-sul1. Four different types of gene cassette arrays were found and a highly prevalent array of dfrA12-orfF-aadA2 gene cassettes was observed. Thirty class 1 integron-positive coagulase-negative staphylococci strains were subjected to Southern hybridization analysis; the result showed that class 1 integrons were located on chromosome, not plasmid. According to the results of SCCmec typing for 30 integron-bearing MRCNS strains, five, 15 and five strains belonged to type I, II and III SCCmec, respectively, and five strains were untypeable. For 23 non-integron-bearing methicillin-resistant coagulase-negative staphylococci strains, four, nine and seven strains belonged to type I, II and III SCCmec, respectively, and three strains were untypeable. None of the strains belonged to type IV or V. Twenty-three coagulase-negative staphylococci isolates of three Staphylococcal species that contained the dfrA12-orfF-aadA2 gene cassette array were phylogenetically unrelated to each other by randomly amplified polymorphic DNA, indicating that the gene cassettes might be disseminated in the clinical strains by a horizontal gene transfer.     

Comparative evaluation of bacteria identification from the Acinetobacter genus using a commercially available API 20NE system, PCR and RFLP techniques

A study was carried out for identification of 50 Acinetobacter strains isolated from various clinical materials. Using classic methods the following species were identifies: Acinetobacter sp. (68%), Acinetobacter baumanii (24%) and Acinetobacter lwofii (8%). In all strains the recA gene was found of 435-500 pz size which confirms their belonging to that genus. Amplification products were digested with restriction enzymes Mbol and HinfI (RFLP) and their detection was carried out on agarose gel by electrophoresis methods, owing to that the arrangement of gene fragment characteristic of each strain was obtained. After careful analysis restriction patterns were obtained corresponding to the following genome species: Acinetobacter baumanii (60%), Acinetobacter sp. 3 (28%) and Acinetobacter lwoffii (12%). The methods of molecular biology made possible a more precise classification of the studied strains according to species. Certain strains determined as Acinetobacter sp. by the API 20NE system were found to be Acinetobacter baumanii, Acinetobacter sp. 3 or Acinetobacter lwofii when determined by the PCR/RFLP method.     

鲍曼不动杆菌整合子分型与耐药性的初步研究

目的了解临床分离的43株鲍曼不动杆菌中Ⅰ类、Ⅱ类、Ⅲ类整合子的流行病学现状及其耐药性。方法用多重PCR方法扩增30株鲍曼不动杆菌Ⅰ类、Ⅱ类、Ⅲ类整合酶基因,用K—B法检测鲍曼不动杆菌的耐药情况。结果Ⅰ类整合子检出率为79．1％（34／43）,未检出Ⅱ、Ⅲ类整合子。结论Ⅰ类整合子阳性的菌株的耐药率较高,显著高于Ⅰ类整合子阴性的菌株,多重PCR是筛查革兰阴性杆菌整合子的有效方法。     

Effects of Carbapenem consumption on the prevalence of Acinetobacter infection in intensive care unit patients

Background

The consumption of carbapenems has increased worldwide, together with the increase in resistant gram negative bacilli. Subsequently, the prevalence of carbapenem-resistant Acinetobacter infections has increased rapidly and become a significant problem particularly in intensive care unit patients. The aim of the present study was to evaluate the changes in the prevalence of Acinetobacter infection by restricting the consumption of carbapenems in intensive care unit patients.

Methods

This study was conducted between May 1, 2011 and February 28, 2013. The amount of carbapenem consumption and the number of patients with multi-drug resistant Acinetobacter baumannii (MDRAB) isolates during the study period were retrospectively obtained from the records of the patients, who were hospitalized in the intensive care unit. The study period was divided into two periods named as: Carbapenem non-restricted period (CNRP) and carbapenem-restricted period (CRP). During CNRP, no restrictions were made on the use of carbapenems. During CRP, the use of carbapenems was not allowed if there was an alternative to carbapenems. Primary Endpoint: MDRAB infection after ICU admission. The definition of nosocomial infections related to Acinetobacter spp. was based on the criteria of the Center for Disease Control (CDC). The correlation between the amount of carbapenem consumption and the number of infections with MDRAB strains between the two periods were evaluated.

Results

During the study period, a total of 1822 patients’ (1053 patients in CNRP and 769 patients in CRP) records were evaluated retrospectively. A total of 10.82 defined daily dose (DDD/100 ICU days) of anti-pseudomonal carbapenem were used in CNRP, and this figure decreased to 6.95 DDD/100 ICU days in CRP. In the 8-month CNRP, 42 (3.98%) MDRAB-related nosocomial infections were detected, and 14 (1.82%) infections were detected in CRP (p?=?0.012).

Conclusion

The prevalence of MDRAB strains isolated in the CNRP was 2.24-fold higher than the prevalence in the CRP. The prevalence of Acinetobacter infections can be reduced by taking strict isolation measures as well as by implementing good antibiotics usage policy.     

主要医院感染病原菌与社区感染株耐药性比较

目的比较主要医院感染(HAI)病原菌与社区感染(CAI)株的耐药性,指导合理使用抗菌药物。方法收集永康市第一人民医院2003年1月至2006年6月所有标本中分离的主要HAI菌及其CAI株,分别统计其药物敏感试验。采用美国Dade Behring Microscan Walkaway 40全自动细菌鉴定及药敏测试仪及其配套药敏鉴定板测定MIC值。全国医院感染监测网软件和χ2统计分析。结果主要HAI菌为前4种革兰阳性(G )菌依次是金黄色葡萄球菌、表皮葡萄球菌、屎肠球菌、溶血葡萄球菌,前5种革兰阴性(G-)菌依次是大肠埃希菌、肺炎克雷伯菌、鲍曼不动杆菌、铜绿假单胞菌、阴沟肠杆菌。HAI株耐药率普遍高于CAI株,不同的细菌耐药率各具特点。不论HAI株还是CAI株,G 菌对万古霉素的耐药率最低,G-菌对亚胺培南的耐药率均较低,且两者耐药率差异均无显著性。金黄色葡萄球菌、大肠埃希菌、肺炎克雷伯菌、鲍曼不动杆菌4种HAI株对大多数抗菌药物的耐药率明显高于CAI株,其余5种细菌HAI株仅对少数抗菌药物的耐药率高于CAI株。结论HAI菌株耐药性比CAI菌株强,临床应区分感染性质合理使用抗菌药物,有针对性控制感染,从而减少抗菌药物的滥用和细菌耐药性的产生。     

Distribution and molecular profiling of class 1 integrons in MDR Acinetobacter baumannii isolates and whole genome-based analysis of antibiotic resistance mechanisms in a representative strain

The class 1 integron is an important driver of the nosocomial dissemination of multidrug-resistant (MDR) bacteria, such as Acinetobacters. In this study, we characterized the gene cassette arrays of class 1 integrons in Acinetobacter baumannii, where the detailed structure of these integrons for 38 clinical strains was analyzed. The results showed that there are three types of gene cassette arrays that are carried by different class 1 integrons, among them the aac(6′)-IId-catB8-aadA1 array was the most prevalent. For detailed analysis of the integron structure, whole genome sequencing was carried out on strain AB16, and it was found that a single integron on its chromosome has a partial Tn21 transposon in its 5′ flanking region and two complete copies of the insertion element IS26 in both the 5′ and 3′ flanking regions, indicating that the integron could be acquired by horizontal gene transfer. Furthermore, there is one resistance island AbaR22, one bla gene containing a transposon, four intrinsic resistant genes and one efflux pump that together confer six types of antibiotic resistance.     

Concerning the sensitivity of Acinetobacter calcoaceticus

The strains of Acinetobacter calcoaceticus, var. anitratus (A. c. a.) were isolated in the nosocomial environment as an opportune pathogen. The therapy of choice may be determined after in vitro tests. Our results show following therapeutical possibilities: beta-lactam antibiotics--cephalosporins of IIIrd generation (cefotaxime), also combinations of antimicrobials have shown good results: amoxycillin or ticarcillin with clavulanic acid. Best synergistic effect was found in combination ticarcillin-amikacin.     

Postsurgical meningitis caused by Acinetobacter baumannii associated with high mortality

Twenty five (25) cases of nosocomial postsurgical meningitis due to Acinetobacter baumannii meningitis were compared to other 146 cases of meningitis after surgery caused by other pathogens. Prior neurosurgical ventriculo-peritoneal shunt insertion and CNS abnormality as well as very low birth weight were significant risk factors for acquisition of Acinetobacter baumannii meningitis. Mortality - 40% among children with nosocomial meningitis was unacceptably high and significantly higher than among meningitis caused by microorganisms other than Acinetobacter baumannii.     

医院内4种主要革兰阴性菌分布及耐药率分析

摘要：目的 分析大连某三甲医院4种主要革兰阴性病原菌的分布及细菌耐药性变迁,为抗菌药物的合理使用提供依据。方法 回顾性分析2012?2015年住院患者中分离的4种革兰阴性菌临床分布及每年的耐药率,利用WHONET 5.6软件进行统计分析。结果 4年分离致病菌28 485株,其中大肠埃希菌2 994株,肺炎克雷伯菌1 375株,鲍曼不动杆菌1 079株,铜绿假单胞菌1 998株,共占分离致病菌总数的26.1%。4种菌对哌拉西林的耐药率最高（37.2%~78.2%）,对头孢哌酮/苏巴坦耐药率最低（6.7%~39.1%）。2012?2015年,大肠埃希菌、肺炎克雷伯菌和铜绿假单胞菌的抗生素耐药率有下降趋势,鲍曼不动杆菌对多数抗生素耐药率都高,且逐年上升。结论 本院4年感染中4种主要革兰阴性菌占主导地位。4种菌对不同抗生素耐药率和变化不同,鲍曼不动杆菌耐药最严重。4种菌对于头孢哌酮/苏巴坦均比较敏感,对哌拉西林耐药严重。在临床治疗中应有针对性加强管理和用药。