Objective To investigate the characteristics and drug sensitivity of carbapenem-resistance Enterobacteriaceae (CRE) infection in our hospital,and study the genotypic analysis of carbapenemase genes.Methods A total of 140 strains of CRE were collected and the epidemiological date were analyzed.CRE were identified by using VITEK 2-Compact bacterial identification system,the antibiotics-sensitivity were tested by kirby-bauer disk susceptibility test,carbapenemase production was detected by the Modified Hodge Test(MHT),and the carbapenemase genes(KPC,IMP,VIM,NDM,OXA-48) were detected by PCR technology.Results Klebsiella pneumoniae was dominated in all CRE,accounting for 70.71％,followed by Enterobacter aerogenes (6.44％) and Enterobacter cloacae (5.71％).CRE samples were mainly isolated from sputum,accounting for 69.29％,followed by urine (12.86％),secretion (4.29％) and blood (4.29％).The most of the patients were from ICU,accounting for 44.29％ and followed by neurosurgery(18.57％) and hematology (5.00％).The drug sensitivity test showed CRE were high resistant to most of β-lactam antibiotics,the resistant rate was higher than 80％,the resistant rate to minocyline and amikacin were 14.78％ and 35.43％,respectively.Modified Hodge test showed 127 strains were positive.PCR and gene sequencing showed 99 strains CRE carrying KPC-2,14 strains carrying IMP-4,2strains carrying IMP-1 and 13strains carrying NDM-1.Conclusion CRE is highly resistant to most antibiotics in our hospital,and the main resistant gene is KPC,followed by IMP and NDM.According to susceptibility test,the hospital should enhance the drug-resistance surveillance and use antibiotics reasonable to control the multi-drug resistance and the prevalence of resistant strains.

目的：为临床治疗铜绿假单胞菌（PAE）感染提供参考。方法分析医院感染的1049株PAE的耐药性。结果 PAE对复方磺胺甲基异恶唑、米诺环素的耐药率为100%；而对多黏菌素B的耐药率为0，其次为阿米卡星（1.3%）、妥布霉素（4.9%）。结论对临床分离的铜绿假单胞菌进行耐药性动态监测，可以指导临床合理使用抗菌药物，及时调整治疗方案，减少医院感染患者的医疗费用，并有效控制和延缓细菌耐药的发生。

马尔尼菲青霉菌（penicillium marneffei ， PM ）是青霉菌中惟一呈温度双相型的致病菌，临床上比较罕见。本病可发生于健康者，但更多见于免疫缺陷或免疫功能抑制者。随着HIV感染者日见增多，马尔尼菲青霉菌病（penicilliosis marneffei ， PSM）报道也逐年增加。该菌造成的感染主要累及单核－巨噬细胞系统，常播散全身，病死率高，是一种严重的深部真菌病。现将本院收治的马尔尼菲青霉菌感染1例报道如下。

目的:分析耐碳青霉烯类肠杆菌科细菌的分布情况及耐药性.方法:收集丽水市中心医院2010年12月～2013年6月临床分离的耐药菌株,采用法国梅里埃Vitek-2 compact细菌鉴定仪鉴定细菌,药物敏感性试验采用纸片扩散法(K-B法).结果:共收集耐药菌株83株,其中肺炎克雷伯菌占68.67％,产气肠杆菌和阴沟肠杆菌各6.02％,大肠埃希菌2.41％.标本来源最常见为痰液(80.72％),其次尿液(8.43％)和分泌物(4.82％),科室分布最多者为重症监护病房(39.76％),其次为神经外科(36.51％).74例患者在检测出耐药菌株前1个月内使用过至少一种抗菌药.有69.88％和31.33％的患者使用过哌拉西林/他唑巴坦和头孢哌酮/舒巴坦,15.66％和10.84％使用过亚胺培南/西司他丁和美罗培南.耐药菌株对多黏菌素B高度敏感,对米诺环素和阿米卡星的耐药率为13.25％和28.92％,对其他抗菌药的耐药率多介于70％ ～ 100％.结论:耐碳青霉烯类肠杆菌科细菌在医院重症监护病房和神经外科相对集中,对多数临床常用抗菌药呈高度耐药,给临床治疗带来严重的挑战.医院应做好院感监测,合理使用抗菌药,从而有效预防和控制耐药菌株的产生和传播.

Objective To determine the antibiotic resistance of clinical isolates of Acinetobacter baumani from intensive care unit (ICU). Methods One hundred and sixty three strains of Acinetobacter baumani were isolated from clinical samples of ICU patients. The isolates were identified by Vitek2- Compact bacterial identification analyzers (Biomerieux). Disk diffusion test (Kirby- Bauer method) was used to determine the antibiotic susceptibility as recommended by CLIS 2012, the changes of antibi-otic resistance were analyzed by WHONET5.4 software. Results There were 143 strains isolated from the sputum specimens (87.73%), followed by urine specimens (5.52%). The resistances to Piperacil in, Ceftazidine, Cefepime, SMZ- TMP, Ciprofloxacin, Piperacil in/tazobactam, levofioxacin, Ticarcil in/clavulanic acid, Imipenem and Meropenem were 93.25%, 93.25%, 92.64%, 92.64%, 92.02%, 92.02%, 92.02%, 92.02%, 89.57% and 89.57%, respectively. The resistance to Cefeperazone/sulbacta and polymyxin B were only 17.18% and 0.0%, respectively. Conclusion The drug resistance rate of clinical isolates Acinetobacter baumani is high in ICU patients.

OBJECTIVE To analyze the distribution and drug resistance of common species of pathogens causing uri-nary tract infections so as to provide guidance for reasonable clinical use of antibiotics and control of nosocomial in-fections .METHODS From Jan 2012 to Dec 2012 ,the urine specimens were collected ,then the identification of iso-lated pathogens was carried out by using automatic bacteria identification system VITEK-2 Compact of BioMérieux ,France;the drug susceptibility testing was performed with the use of K-B disk diffusion method ,and the data were statistically analyzed with the use of WHONET 5 .5 software .RESULTS A total of 573 strains of pathogens have been isolated with the positive rate of 30 .62% ,including 380 (66 .32% ) strains of gram-negative bacteria ,138 (24 .08% ) strains of gram-positive bacteria ,and 55 (9 .60% ) strains of fungi;Escherichia coli was the predominant species of gram-negative bacteria ,Enterococcus was dominant among the gram-positive bacteria , and Candida albicans was the predominant species of fungi .The gram-negative bacteria were highly resistant to ampicillin ,levofloxacin ,sulfamethoxazole-trimethoprim ,and gentamicin but were highly susceptible to carbapene-ms with the drug resistance rate less than 6 .0% ;the gram-positive bacteria were 100 .0% susceptible to vancomy-cin ,linezolid ,and nitrofurantoin but were highly resistant to levofloxacin ,sulfamethoxazole-trimethoprim ,peni-cillin ,and gentamicin .CONCLUSION E .coli and Enterococcus are the predominant species of pathogens causing the urinary tract infections .The drug resistance of such pathogens is increasing year by year ,therefore ,it is neces-sary for the hospital to reasonably use antibiotics based on the results of urine culture and the drug susceptibility testing .

目的:了解医院儿科住院患者痰液病原菌类型及耐药性情况.方法:分离儿科病房送检痰液标本病原菌,采用Vitek2-Compact系统鉴定细菌,K-B纸片扩散法进行药物敏感试验,分析常见病原菌耐药性.结果:2010～ 2012年共送检标本2 814份,检出病原菌1 005株,检出率35.71％.其中革兰阳性菌335株,以金黄色葡萄球菌、表皮葡萄球菌、肺炎链球菌为主.革兰阴性菌625株,以流感嗜血杆菌、肺炎克雷伯菌、副流感嗜血杆菌、大肠埃希菌和卡他莫拉菌常见.真菌45株,以白色假丝酵母菌和热带假丝酵母菌常见.葡萄球菌对青霉素和红霉素耐药性较高,肺炎链球菌对红霉素和复方磺胺甲嗯唑高度耐药.大肠埃希菌和肺炎克雷伯菌对头孢菌素类的耐药率约60％,嗜血杆菌对氨苄西林、阿奇霉素、复方磺胺甲嗯唑存在不同程度的耐药.卡他莫拉菌对青霉素和氨苄西林高度耐药.结论:儿童下呼吸道感染以革兰阴性菌为主,临床上应及时检测,应根据本院病原菌谱及药敏情况合理选择抗菌药.

One highly multi-drug-resistant, mucus-producing and foul-smelling strain of Klebsiella ozaenae was isolated from a patient in the ICU of a Chinese tertiary hospital. MICs of several clinical antimicrobials against the strain were obtained using the Vitek-2 Compact System with AST-GN13 cards and resistance genes were evaluated by PCR and gene sequencing. The strain was resistant to most of the β-lactams and quinolones tested and carried several antibiotic resistance genes, including bla(KPC-2), bla(TEM-98), bla(CTX-M-3), bla(SHV-26) and qnrS. To our knowledge, this is the first report of β-lactam and quinolone resistance genes co-existing in a K. ozaenae strain in China.

从浙江丽水市中心医院分离到无重复泛耐药克雷伯菌7株:5株肺炎克雷伯菌(K1-K5)、1株臭鼻克雷伯菌(K6)、1株解鸟氨酸克雷伯菌(K7). 菌株鉴定及药敏:取患者痰液标本在血平板上,37℃,18～24 h生长分离单个菌落.采用Vitek2 -compact鉴定仪经GNI卡鉴定为克雷伯菌.取M-H平板上37℃,18 h生长的单个菌落,采用鉴定仪配套的GN13卡鉴定菌株药敏.结果所有菌株对亚胺培南、厄他培南、头孢他啶、头孢吡肟等碳青霉烯类、头孢类抗生素耐药,且均对氨苄西林/舒巴坦、哌拉西林/他唑巴坦、复方新诺明、氨曲南、呋喃妥因耐药;除解鸟氨酸克雷伯菌(K7)外,其他菌株同时对左氧氟沙星、环丙沙星等喹诺酮类抗生素耐药.

Objective To study the extensively drug resistant mechanisms of Klebsiella pneumoniae and discuss the clinical treatment strategies. Methods The isolate was identified by Vitek-compact system,the drug susceptibility test was conducted by K-B method,and KPC-2 gene was detected by Hodge test and polymerase chain reaction (PCR),then sequenced,and the treatment of the patient was investigated. Results The strain was resistant to all antibiotics used in routine drug susceptibility test except polymyxin and minocycline. The inhibition zone diameter of fosfomycin added later was 20mm. In synergistic drug susceptibility test,no synergistic antimicrobial activity was found between polymyxin,minocycline,fosfomycin and imipenem. Hodge test was positive; KPC-2 gene was detected by PCR and the sequence was consistent with AF297554 in GenBank. After treatment for one month,no extensively drug resistant Klebsiella pneumoniae strain was detected from the patients. Conclusion It is necessary to strengthen the monitoring of extensively drug resistant Klebsiella pneumoniae and improve the awareness of extensively drug resistant bacteria to facilitate the prevention and treatment of infectious disease.

新生儿败血症主要病原菌为葡萄球菌,其中耐甲氧西林葡萄球菌(MRS)对临床常用青霉素类和头孢类抗生素耐药,对其快速鉴定有助于合理选择治疗药物.本文介绍直接用新生儿血培养阳性培养瓶中的上层液体做头孢西丁纸片扩散法鉴定MRS的方法.

AdeABC efflux pump in a total of 50 Acinetobacter baumannii strains were investigated and the role in contributing to hydrolysis carbapenems were further analyzed. All strains were divided into 3 groups according to their susceptibilities to Imipenem, Amikacin, Minocycline and Levofloxacin: Group A (22 imipenem-resistant strains), B (13 isolates which were imipenem-sensitive but resistant to at least one of the other three antibiotics) and C (15 isolates, sensitive to all the antimicrobials). Five gene types were observed according to REP-PCR and 39 isolates were included in the main one. Only one isolate (A2) was positive for efflux pump phenotype. All strains were positive for blaOXA51-like and AdeB gene and negative for blaOXA24, blaOXA58, VIM, IPM, and SIM-1. blaOXA23 were detectable merely in the 22 imipenem-resistant strains. 24 isolates from the same REP type representing for the 3 groups were selected for quantitative analysis of adeB expression. Compared with the mean level of Group B, only A2 expressed apparently higher (2.2 fold). The induction effects of imipenem and meropenem were analyzed as well. Compared with their freely grew isogenic counterparts, up regulated expression was observed only in 2 isolates under the pressure of imipenem and none in all under meropenem. These data indicate that AdeABC efflux pump play a less important role in A. baumannii against carbapenems. Key words: Acinetobacter baumannii, Carbapenem resistance, AdeABC efflux pump, adeB gene.

万古霉素是一种糖肽类抗菌药物,在治疗葡萄球菌感染中具有重要作用,尤其是在治疗耐甲氧西林葡萄球菌引起的各类感染中效果显著[1].虽然耐万古霉素葡萄球菌鲜见报道[1-2],但随着糖肽类抗菌药物使用增加,葡萄球菌对万古霉素的敏感性下降,给临床治疗带来了困难和挑战[3-4],故正确检测葡萄球菌对万古霉素敏感性对治疗有重要意义.

Objective:To study the etiology of orthopaedic infection and the drug resistance characteristic of infective bacteria in order to direct clinic using anti-infective drug reasonably.Methods:The germ composition of 2054 orthopaedic specimens and their drug-resistance character were investigated.Results:Totally 889 infected specimens were detected,and 986 isolates were identified,including 376 Gram-positive bacilli,accounted for 38.1%,545 Gram-negative bacilli,accounted for 55.3%,and 65 fungal strains,accounted for 6.6%,respectively.The main pathogens in wound infections were Staphylococcus aureus,coagulase-negative-staphylococcus(CNS) and Pseudomonas aeruginosa and their drug-resistance was various.Conclusion:Bacteria causing orthopaedic infection are widespread,and their drug-resistance character is various.Successful treatment would be carried out according to the result of germiculture and antimicrobial susceptibility testing.

Objective:To analyze the relationship between imipenem inhibition zone and KPC type Carbapenemase in Klebsiella pneumoniae.Methods:K.pneumoniae strains were separated in two groups according to ESBLs.Imipenem inhibition zone were measured with disc dilution method,while KPC gene was detected by PCR.Results:Imipenem inhibition zone in 81 ESBLs positive and 101 ESBLs negative strains were 27.92±0.18 cm and 28.28±0.11 cm,respectively,according to statistical method.But there was no significant discrepancy between the two groups(P>0.05).KPC gene were detected in 2 ESBLs positive strains with the inhibition zone of 20 cm and 19.5 cm respectively.Conclusion:KPC type Carbapenemase may contribute to imipenem resistant in K.pneumoniae with the inhibition zone <= 20 cm.

目的:分析丽水市中心医院鲍曼不动杆菌耐药特征及blaOXA23基因携带情况,指导临床合理应用抗生素.方法:经VITEK2-compact微生物系统鉴定,收集丽水市中心医院2008年3月至2009年6月从临床分离的鲍曼不动杆菌62株.统计其科室来源,以K-B法检测菌株对抗生素的耐药性,并利用PCR扩增法检碳青酶烯酶耐药基因blaOXA23.结果:鲍曼不动杆菌检出率前两位是脑外科和ICU病房,分别占32.3%和27.4%.多重耐药鲍曼不动杆菌阳性率最高的是ICU,为70.6%(12/17);其次为脑外科,为35.0%(7/20).亚胺培南、美洛培南、头孢他啶、头孢吡肟、替卡西林/克拉维酸、哌拉西林/他唑巴坦、头孢哌酮/舒巴坦、多粘菌素耐药率分别为33.9%、38.9%、61.3%、53.3%、56.1%、45.2%、16.9%和0.0%.blaOXA23总检出率35.5%,亚胺培南耐药鲍曼blaOXA23检出率100.0%,敏感鲍曼均未检出.结论:鲍曼不动杆菌分布广泛,并且多重耐药性严重;blaOXA23基因可能是耐碳青酶烯类抗生素的主要原因.建议加强鲍曼不动杆菌耐药性监测及耐药机制的研究.

从老年慢性阻塞性肺疾病患者痰标本中分离到1株罕见的泛耐药黏液型鲍曼不动杆菌(slime-production pandmg-re-sistant A. baumannii,SPAB),现报告如下.

Objective To review the status of diagnosis and treatment for invasive fungal pulmonary infections(IFPI)in Lishui Central Hospital.Methods The clinical data of 79 patients with IFPI were retrospectively analyzed.Results The diagnostic status could be classified ills follows:6 eases had confirmed diagnosis,30 had clinical diagnosis,35 had suspected diagnosis and 8 misdiagnosed.The treatments were all effeetive in 6 COnfirmed cases;in 30 clinically diagnosed cases,6 were eriective.21 were inefiective and 3 died;in 35 suspected cases.3 were effective.25 were iHefieetive and 7 cflses did not receive antifungal treatment.Aspergillus and Cryptococcus pulmonary infections were predominant in confirmed cases.and the antifungal treatment lasted for 3 to 6 months.Conclusion Diagnosis and treatment for IFPI need to be improved.

OBJECTIVE To detect genes sul1 and sul2 in Stenotrophomonas maltophilia,and their relationship to drug resistance to trimethoprim-sulfamethoxazole(SXT).METHODS K-B was carried out to detect the drug resistance to SXT of S.maltophilia;minimal inhibitory concentration(MIC) was measured with micro broth dilution method.Genes sul1 and sul2 were amplified by PCR.RESULTS Eight isolates(7.8%) showed resistance to trimethoprim-sulfamethoxazole,sul1 Was positive in four isolates in which one contained sul2 also.Four isolates showed high MIC to SXT.CONCLUSIONS Genes sul1 and sul2 of S.maltophilia are associated with the high drug resistance to SXT.

Objective:To evaluate the reliability of VITEK2-compact extended-spectrum-β-lactamases test in Escherichia coli and Klebsiella pneumoniae.Methods:Detection of ESBLs in Escherichia coli and Klebsiella pneumoniae with phenotype confirmatory test,meanwhile,to mate detection strains ESBLs by VITEK2-compact analyzer.To compare the ESBLs detection rates and detection time;presuming types of ESBLs according to resistant phenotype to cefotaxim and ceftazidime,and comparing the coincidence in two methods.Results:The coincidence was 97.37% with two methods ESBLs test in 78 strains Escherichia coli.and in 28 strains Klebsiella pneumoniae that was 100%,average detection time was 5.5 hours with VITEK2-compact analyzer.In contrast,that was 17.0 hours.The coincidence of ESBLs types detection in two methods was 100%.Conclusion:ESBLs detection of Escherichia coli and Klebsiella pneumoniae with VITEK2-compact is reliable and rapid.