耐甲氧西林表皮葡萄球菌及耐药性检测和同源性分析
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摘要
耐甲氧西林金黄色葡萄球菌(MRSA)目前成为医院感染的重要病原菌之一,很多文献报道其发生率持续上升。而凝固酶阴性的耐甲氧西林表皮葡萄球菌(MRSE)的发生率则更高,有报道其发生率为80%或以上。MRSE不仅对甲氧西林耐药,还对其他多种抗菌药物耐药,具耐多药特征,已是临床抗感染化疗的难题之一。本研究对安徽省部分医院临床分离的表皮葡萄球菌进行检测,检出其中的耐甲氧西林表皮葡萄球菌,了解本地区MRSE的检出率,比较两种检测方法,对检出的MRSE进行耐药性检测,为临床防治MRSE感染及合理使用抗生素提供实验依据;并对MRSE进行分子生物学研究,这对于细菌性传染病的监测,传染源追踪、传播途径调查和识别等爆发调查有着非常重要的意义。
目的
了解本地区MRSE的检出率,比较两种检测方法;对检出的MRSE进行耐药性检测,了解本地MRSE耐药的特点;并对MRSE菌株的同源性进行研究。为临床合理用药和耐药菌的防治提供参考依据。
方法
采用头孢西丁纸片扩散法及mecA基因PCR法检测出MRSE菌株,并对两种检测方法加以比较。以PCR方法检测出mecA基因作为检测MRSE菌株的金标准,了解头孢西丁纸片扩散法检测MRSE菌株的灵敏度及特异度。采用琼脂稀释法测定分离出的表皮葡萄球菌对17种常用抗生素的最低抑菌浓度(MIC)。以肠道细菌基因间重复一致序列(enterobacterial repetitive integenic consensus,ERIC)为引物结合位点,PCR扩增MRSE基因,并进行同源性分析。
结果
128株临床分离表皮葡萄球菌中,用头孢西丁纸片法及PCR法对MRSE检出率分别为79.69%(102/128)和85.16%(109/128),两种方法对MRSE检出率差异无统计学意义。以PCR法检出mecA基因作为判断MRSE的金标准,头孢西丁纸片法灵敏度为88.07%、特异度为68.42%、符合率为88.16%。MRSE对多种抗菌药物耐药显著;除对β-内酰胺类明显耐药外,对红霉素和克林霉素的耐药率均高达98.17%,对氟喹诺酮类(左氧氟沙星和环丙沙星)的耐药率亦超过80%。MRSE对万古霉素、替考拉宁100%敏感,未发现耐药菌株。利用ERIC-PCR检测MRSE的同源性,结果显示部分MRSE菌株间谱型相同。
结论
头孢西丁纸片法是筛选和确认MRSE菌株的一种可靠、简便的方法。MRSE为多重耐药菌株,耐药形势严峻,部分菌株间存在克隆传播现象,临床必须加强细菌耐药性监测。
Along with the wide application antimicrobial agents,staphylococci have become an important source of nosocomial infection.The resistance of methicillin resistant staphylococcus epidermidis(MRSE) has increased even more than the resistance rate of MRSA.And caused more and more difficulities for clinical therapy.We evaluated the two methods for detection of MRSE,and analysis the resistance of staphylococcus epidemidis isolated from 20 hospitals in Anhui from January to December in 2005.Based on the study of molecular biological mechanism and molecular epidemiology,test the gene of MRSE.The results of present study may contribute to clinical therapy and molecular epidemiology of MRSE.
Objective:Investigating the resistant of MRSE,evaluating the two methods for detetion of MRSE.Investigating the homology in MRSE isolates.
Methods:MRSE were detected by cefoxitin disk diffusion,as well as PCR amplification for the mecA gene,respectively.128 strains of S.epidermidis isolates were tested for minimus inhibitory concentration(MIC) to 17 kinds of antimicrobial agents with agar dilution method.Taking enterobacterial repetitive integenic consensus(ERIC) as primer binding site,analysis the homology in MRSE by PCR.
Results:The rates of MRSE detected by cefoxitin disk diffusion as well as PCR amplification for the mecA gene were79.69%and 85.16%,respectively,and the result of two groups had no difference(P＞0.05).Cefoxitin disk diffusion method of the specificity and sensitivity were higher enough to replace mecA PCR in clinical work.Except penicillin-G,resistance rate of MRSE to erythromycin and clindamycin were high too.And all strains were sensitive to vancomycin.Some isolates of MRSE have same fingerprint of genotype.
Conclusions:Cefoxitin disk diffusion method can be used reliably in detecting and conforming MRSE.Resistance in MRSE was getting more and more serious,and some MRSE strains had clonal dissemination.So detection and surveillance of them should be emphasized.
目的
了解本地区MRSE的检出率,比较两种检测方法;对检出的MRSE进行耐药性检测,了解本地MRSE耐药的特点;并对MRSE菌株的同源性进行研究。为临床合理用药和耐药菌的防治提供参考依据。
方法
采用头孢西丁纸片扩散法及mecA基因PCR法检测出MRSE菌株,并对两种检测方法加以比较。以PCR方法检测出mecA基因作为检测MRSE菌株的金标准,了解头孢西丁纸片扩散法检测MRSE菌株的灵敏度及特异度。采用琼脂稀释法测定分离出的表皮葡萄球菌对17种常用抗生素的最低抑菌浓度(MIC)。以肠道细菌基因间重复一致序列(enterobacterial repetitive integenic consensus,ERIC)为引物结合位点,PCR扩增MRSE基因,并进行同源性分析。
结果
128株临床分离表皮葡萄球菌中,用头孢西丁纸片法及PCR法对MRSE检出率分别为79.69%(102/128)和85.16%(109/128),两种方法对MRSE检出率差异无统计学意义。以PCR法检出mecA基因作为判断MRSE的金标准,头孢西丁纸片法灵敏度为88.07%、特异度为68.42%、符合率为88.16%。MRSE对多种抗菌药物耐药显著;除对β-内酰胺类明显耐药外,对红霉素和克林霉素的耐药率均高达98.17%,对氟喹诺酮类(左氧氟沙星和环丙沙星)的耐药率亦超过80%。MRSE对万古霉素、替考拉宁100%敏感,未发现耐药菌株。利用ERIC-PCR检测MRSE的同源性,结果显示部分MRSE菌株间谱型相同。
结论
头孢西丁纸片法是筛选和确认MRSE菌株的一种可靠、简便的方法。MRSE为多重耐药菌株,耐药形势严峻,部分菌株间存在克隆传播现象,临床必须加强细菌耐药性监测。
Along with the wide application antimicrobial agents,staphylococci have become an important source of nosocomial infection.The resistance of methicillin resistant staphylococcus epidermidis(MRSE) has increased even more than the resistance rate of MRSA.And caused more and more difficulities for clinical therapy.We evaluated the two methods for detection of MRSE,and analysis the resistance of staphylococcus epidemidis isolated from 20 hospitals in Anhui from January to December in 2005.Based on the study of molecular biological mechanism and molecular epidemiology,test the gene of MRSE.The results of present study may contribute to clinical therapy and molecular epidemiology of MRSE.
Objective:Investigating the resistant of MRSE,evaluating the two methods for detetion of MRSE.Investigating the homology in MRSE isolates.
Methods:MRSE were detected by cefoxitin disk diffusion,as well as PCR amplification for the mecA gene,respectively.128 strains of S.epidermidis isolates were tested for minimus inhibitory concentration(MIC) to 17 kinds of antimicrobial agents with agar dilution method.Taking enterobacterial repetitive integenic consensus(ERIC) as primer binding site,analysis the homology in MRSE by PCR.
Results:The rates of MRSE detected by cefoxitin disk diffusion as well as PCR amplification for the mecA gene were79.69%and 85.16%,respectively,and the result of two groups had no difference(P＞0.05).Cefoxitin disk diffusion method of the specificity and sensitivity were higher enough to replace mecA PCR in clinical work.Except penicillin-G,resistance rate of MRSE to erythromycin and clindamycin were high too.And all strains were sensitive to vancomycin.Some isolates of MRSE have same fingerprint of genotype.
Conclusions:Cefoxitin disk diffusion method can be used reliably in detecting and conforming MRSE.Resistance in MRSE was getting more and more serious,and some MRSE strains had clonal dissemination.So detection and surveillance of them should be emphasized.
引文
1、徐小平,高霞,池小霞等.葡萄球菌属和肠球菌属耐药性监测研究.中华医院感染学杂志,2006,16(3):324-327.
2、邵亚娟,乔惠平,吴玉莲.医院感染常见细菌分布与耐药性分析.中国感染控制杂志,2006,5(2):153-156.
3、陈小会,金月英.杭州市医院感染病原菌分布及耐药性分析.浙江预防医学,2005,17(8):31-32.
4、 Rupp ME,Archer GL.Coagulase-negative staphylococci:pathogens associated with medical progress[J].Clin Infect Dis,1994,19(2):231-243.
5、张永标,梁彩倩,张扣兴等.凝固酶阴性葡萄球菌谱与耐药谱调查.中国微生态学杂志,2006,18(5):395-396.
6、牛其昌,范春,王英.1999-2003年我院表皮葡萄球菌耐药性变化的分析.中国医院用药评价与分析,2005,5(1):53-55.
7、 Raad I,Alrahwan A,Rolston K.Staphylococcus epidemidis:resistance and need for alternative agents[J].Clin Infect Dis,1998,26(5):1182-1187.
8、王箭,罗军.表皮葡萄球菌所致医院感染的分布及耐药趋势.徐州医学院报,2005,25(4):338-340.
9、贾杰主编.感染病学:新疾病、新理论、新技术.第一版.成都:四川科学技术出版社,2001:978-988.
10、 Schierholz JM,Beuth J.Implant infections:a heaven for opportunistic becteria [J].J Hosp Infect,2001,49(2):87-93.
11、 Vuong C,Otto M.Staphylococcus epidermidis infections.Microbes Infect,2002,4(4):481-489.
12、 Bergamini TM,Corpus RA Jr,McCurry TM,et al.Immunosuppression augments growth of adherent staphylococcus epidermidis[J].Arch Surg,1995,130(12):1345-1350.
13、唐开洪,宋鹏,凌月明等.临床常见病原菌分布及耐药性趋势分析。实用医技杂志,2003,10(6):563-564.
14、熊自忠,李涛,徐元宏.临床分离耐甲氧西林表皮葡萄球菌的耐药性监测.中国医师杂志,2005,7(2):262-263.
15、 Oliver DC,de Lebcastre H.Multiplex PCR strategy for rapid identification of structural typesand variants of the mec element in methicillin-resistant staphylococcus aureus.Antimicrob Agents Chemotherary,2002,46(7):2155-2614.
16、 Performance standards for antimicrobial susceptibility testing,fifteenth informational supplement S.CLSI,2005,M 100-S15 44-50.
17、 Berger-Bachi B,Rohrer S.Factors influencing methicillin resistance in staphylococcus.Arch Microbiol,2002,178(3):165-171.
18、 Predari SC,Ligozzi M,Fontana R.Genotypic identification of methicillin-resistant cogulase-negative staphylococci by polymerase chain reaction.Antimicrob Agents Chemotherary.1991,35(12):2568-2573.
19、 Hoffmann E,Stech J,Webster RG,et al.Universalprimer set for the full-length amplification of all influenza aviruses.J Arch Virol,2001,146(12):2275.
20、 Nishi J,Miyanohara H,Nakajima T,et al.Molecular typing of the methicillin resistance determinant(mec) of clinical strains of staphylococcus based on mec hypervariable region length polymorphisms.Lab Clin Med,1995,126(1):29-35.
21、 Schwarz S,Kehrenberg C,Ojo KK,et al.staphylococcus sciuri gene erm(33),encoding inducible resistance to macrolides,lincosamides,and streptogramin B antibiotics,is a product of recombination between erm(C) and erm(A).Antimicrob Agents Chemotherapy,2002,46(2):3621-3623.
22、孙保良,李之光,张敏.葡萄球菌对大环内酯类的耐药研究.中华检验医学杂志,2006,29(5):459.
23、 Patel MV,De Souza NJ,Gupte SV,et al.Antistaphylococcal Activity of WCK 771,a tricyclic fluoroquinolone,in animal infection models.Antimicrob Agents Chemotherapy,2004,48(12):4754-4751.
24、何礼贤.耐药革兰阳性球菌的医院流行病学及感染控制.国外医药抗生素分册,2003,24(3):103-107.
25、 Archer GL,Molecular epidemiology of multiresistant staphylococcus[J].J Antimcrob Agents Chemother,1988,21(supplc):133.
26、张青,雷呈祥,赵旭.表皮葡萄球菌生物膜形成分子机制的研究进展.微生物学报,2003,43(5):681-685.
27、 Vuong C,Voyich J M,Fischer ER,et al.Polysaccharide intercellular adhesin (PIA) protects staphylococcus epidermidis against major components of the human innate immune system.Cell Microbiol,2004,6(3):269-275.
28、李敏,高谦,王建平等.表皮葡萄球菌附属基因调节因子对生物膜形成的调节作用.中华检验医学杂志,2005,28(11):1172-1177.
29、 Yao Y,Vuong C,Kocianova S,et al.Characterization of the staphylococcus epidermidis accessory-gene regulator reponse:quorum-sensing regulation of resistance to human innate host defense.J Infect Dis,2006,193(6):841-848.
30、李家泰,Allan JW,扬敏.中国细菌耐药监测研究[J].中华医学杂志,2001,81(1):8-15.
31、刑铭友,田德英,刘莉娜.2004-2005同济医院表皮葡萄球菌耐药性分析.华中科技大学学报(医学版),2007,36(5):677-680.
32、赵旭,张婴元.表皮葡萄球菌感染防治.中国抗感染化疗杂志,2003,3(5):318-320.
1、刑铭友,田德英,刘莉娜.2004-2005年同济医院表皮葡萄球菌耐药性分析.华中科技大学学报(医学版),2007,36(5):677-680.
2、赖燕菲,吴瑶,盛吉芳等.256例表皮葡萄球菌医院感染分析.浙江大学学报(医学版),2000,29(5):223-226.
3、 Versalovi CJ,Koeuth T,Lupski J R.Distribution of repetitive DNA sequences in eubecteria and application to fingerprinting of bacterial genomes[J].Nucleic Acids Res,1991,19(24):6823-6831.
4、 Silbert S,Hollis R J.Evaluation of three molecular techniques for nonfermetative Gram-negative bacill.Infect Control Hosp Epidemiol,2004,25(10):847-851.
5、 Gillings M,Holley M.Repetitive element PCR fingerprinting(rep-PCR) using enterobacterial repetitive intergenic consensus(ERIC) primers is not necessarily directed at ERIC elements[J].Letters in Applied Microbiology,1997,25(1):17-21.
6、陈迎春,曹又方,赵立平.大肠杆菌MG1655菌株ERIC-PCR图谱主带序列组成分析[J].微生物学通报,2002,29(6):28-32.
7、 Dalla-Costa L M,Irino K,Rodrigues J,et al.Characterisation of diarrhoeagenic Escherichia coli clones by ribotyping and ERIC-PCR[J].Journal of Medical Microbiology,1998,47(3):227-234.
8、黄革,董婷,侯铁英等.重复序列引物聚合酶链反应追踪金黄色葡萄球菌所致医院感染.中国感染与化疗杂志,2006,6(4):260-262.
9、 Debast SB,Meis JFGM,Melchers WJ,et al.Use of interrepeat PCR fingerprinting to investigate an Acinetobacter baumannii out-break in an intensive care unit.Scan J Infect Dis,1996,28(2):577-581.
10、 Szewcjyk EM,Piltrowski A,Rojalska M.Predominant staphylococci in the intensive care unit of a pediatric hospital[J].Hosp Infect,2000,45(2):145-154.
11、李胜利,张婴元,吴菊芳等.葡萄球菌医院感染的调查研究[J].中华医 院感染学杂志,1998,8(1):65-67.
12、熊自忠,李涛,徐元宏.临床分离耐甲氧西林表皮葡萄球菌的耐药性监测.中国医师杂志,2005,7(2):262-263.
13、 Vuong C,Otto M.Staphylococcus epidermidis infections.Microbes Infect,2002,4(4):481-489.
14、唐开洪,宋鹏,凌月明等.临床常见病原菌分布及耐药趋势分析.实用医技杂志,2003,10(6):563-564.
15、何礼贤.耐药革兰氏阳性球菌的医院流行病学及感染控制.国外医药抗生素分册,2003,24(3):103-107.
16、王海燕,梁慧,崔云龙等.重复片段PCR检测葡萄球菌DNA指纹在医院感染中的分析.中华医院感染学杂志,2003,13(6):514-516.
17、李雅卿,黄卫红.耐甲氧西林金黄色葡萄球菌医院感染分析[J].中华医院感染杂志,2002,12(2):151-152.
18、 Hulton CS,Higgins CF,Shap PM.ERIC sequences:a novel family of repetitive elements in the genomes of Escherchia coli,Salminella typhimurium and other entrobacteria.Mol Microbiol,1991,5(4):825-834.
19、 Jersek B,Tcherneva E,Rijpens N,et al.Repetitive element sequence-based PCR for species and strain discrimination in the genus Listeria.Lett Appl Microbiol,1996,23(1):55-60.
20、李艳琴,申泉,刘彬彬等.番茄内生菌分离及其ERIC-PCR指纹图谱分析.微生物学通报,2003,30(5):89-93.
21、 De Bruijin F J.Use of repetitive(repetitive extragenic palindromic and enterobacterial repetitive intergeneic consensus) sequences and the polymerase chain reaction to fingerprint the genomes of Rhizobium meliloti iaolates and other soil bacteria.Appl Environ Microbiol,1992,58(7):2180-2187.
22、 Di Giovanni GD,Watrud LS,Seidler RJ,et al.Fingerprinting of mixed bacterial strains and BIOLOG grem-negative(GN) substrate communities by enterobacterial repetitive intergenic consensus sequence- PCR (ERIC-PCR).Curr Microbiol,1999,38(4):217-223.
23、高平平,赵立平.微生物群落结构探针杂交评价不同培养基从活性污泥分离优势菌群的能力[J].微生物学报,2003,43(2):264-270.
24、李艳琴,孙永艳,崔丽芳等.ERIC-PCR在人工混合菌体系中的检出灵敏度.微生物学通报,2004,31(4):61-64.
25、李犹平,倪学勤.ERIC-PCR技术特点及其应用.安徽农业科学,2007,35(18):5358-5360.
26、晏群,陈丽华,刘文恩等.分子生物学分型技术ERIC-PCR的建立.实用医学杂志,2003,19(6):688-689.
1、 erformance standards for antimicrobial susceptibility testing;fifteenth informational supplement S.CLSI,2005,M 100-S 15 44-50.
2、 Vuong C,Otto M.staphylococcus epidermidis infections.Microbes Infect,2002,4(4):481-489.
3、唐开洪,宋鹏,凌月明等.临床常见病原菌分布及耐药趋势分析.实用医技杂志,2003,10(6):563-564.
4、 熊自忠,李涛,徐元宏.临床分离耐甲氧西林表皮葡萄球菌的耐药性监测.中国医师杂志,2005,7(2):262-263.
5、 何礼贤.耐药革兰氏阳性球菌的医院流行病学及感染控制.国外医药抗生素分册,2003,24(3):103-107.
6、 陈智鸿,吕晓菊,范昕建等.金黄色葡萄球菌耐药性监测.中国抗感染化疗杂志,2003,3(1):13-15.
7、 赖燕菲,吴瑶,盛吉芳等.256例表皮葡萄球菌医院感染分析.浙江大学学报(医学版),2000,29(5):223-226.
8、 刑铭友,田德英,刘莉娜.2004-2005年同济医院表皮葡萄球菌耐药性分析.华中科技大学学报(医学版),2007,36(5):677-680.
9、 朱德妹,汪复,张婴元.2004年上海地区细菌耐药性监测.中国抗感染化疗杂志,2005,5(4):195-200.
10、余红,李强.四年中临床分离的表皮葡萄球菌耐药性分析.实用全科医学,2005,3(4):366-367.
11、王箭,罗军.表皮葡萄球菌所致医院感染的分布及耐药趋势.徐州医学院学报,2005,25(4):338-340.
12、周桂华,尹有宽.导管相关性感染.国外医学内科学分册,1997,24(6):235-237.
13、 Barie PS.Antibiotic-resistant gram-positive cocci:implications for surgical practice.World J Surg,1998,22(2):118-126.
14、 Rupp ME,Archer GL.Coagulase-negtive staphylococci:pathogen associated with medical progress.Clin Infect Dis,1994,19(2):231-243.
15、 Lim D,Strynadka NC.Structural basis for the beta lactam resistance of PBP2α from methicillin-resistant staphylococcus aureus.Nat Struct Biol,2002,9(11):870-876.
16、 Pagliero E,Cgesnel L,Hopkins J,et al.Biochemical characterization of strephylococcus pneumonia penicillin-binding protein 2b and its implication in beta-lactam resistance.Antimicrob Agents Chemotheary,2004,48(5):1848-1855.
17、陈智鸿,吕晓菊,范昕建等.金黄色葡萄球菌耐药性监测.中国抗感染化疗杂志,2003,3(1):13-15.
18、 Schierholz JM.Beuth J.Implant infections:a heaven for opportunistic becteria [J].J Hosp Infect,2001,49(2):87-93.
19、 Vuong C,Voyich J M,Fischer ER,et al.Polysaccharide intercellular adhesin (PIA) protects staphylococcus epidermidis against major components of the human innate immune sustem.Cell Microbiol,2004,6(3):269-275.
20、江娟,孙景勇,欧元祝等.医源性表皮葡萄球菌ica操纵子转录水平对生物膜表型的影响.上海医学,2006,29(1):40-44.
21、李敏,高谦,王建平等.表皮葡萄球菌附属基因调节因子对生物膜形成的调节作用.中华检验医学杂志,2005,28(11):1172-1177.
22、 Yao Y,Vuong C,Kocianova S,et al.Characterization of the staphylococcus epidermidis accessory-gene regulator response:quorum-sensing regulation of resistance to human innate host defense.J Infect Dis,2006,193(6):841-848.
2、邵亚娟,乔惠平,吴玉莲.医院感染常见细菌分布与耐药性分析.中国感染控制杂志,2006,5(2):153-156.
3、陈小会,金月英.杭州市医院感染病原菌分布及耐药性分析.浙江预防医学,2005,17(8):31-32.
4、 Rupp ME,Archer GL.Coagulase-negative staphylococci:pathogens associated with medical progress[J].Clin Infect Dis,1994,19(2):231-243.
5、张永标,梁彩倩,张扣兴等.凝固酶阴性葡萄球菌谱与耐药谱调查.中国微生态学杂志,2006,18(5):395-396.
6、牛其昌,范春,王英.1999-2003年我院表皮葡萄球菌耐药性变化的分析.中国医院用药评价与分析,2005,5(1):53-55.
7、 Raad I,Alrahwan A,Rolston K.Staphylococcus epidemidis:resistance and need for alternative agents[J].Clin Infect Dis,1998,26(5):1182-1187.
8、王箭,罗军.表皮葡萄球菌所致医院感染的分布及耐药趋势.徐州医学院报,2005,25(4):338-340.
9、贾杰主编.感染病学:新疾病、新理论、新技术.第一版.成都:四川科学技术出版社,2001:978-988.
10、 Schierholz JM,Beuth J.Implant infections:a heaven for opportunistic becteria [J].J Hosp Infect,2001,49(2):87-93.
11、 Vuong C,Otto M.Staphylococcus epidermidis infections.Microbes Infect,2002,4(4):481-489.
12、 Bergamini TM,Corpus RA Jr,McCurry TM,et al.Immunosuppression augments growth of adherent staphylococcus epidermidis[J].Arch Surg,1995,130(12):1345-1350.
13、唐开洪,宋鹏,凌月明等.临床常见病原菌分布及耐药性趋势分析。实用医技杂志,2003,10(6):563-564.
14、熊自忠,李涛,徐元宏.临床分离耐甲氧西林表皮葡萄球菌的耐药性监测.中国医师杂志,2005,7(2):262-263.
15、 Oliver DC,de Lebcastre H.Multiplex PCR strategy for rapid identification of structural typesand variants of the mec element in methicillin-resistant staphylococcus aureus.Antimicrob Agents Chemotherary,2002,46(7):2155-2614.
16、 Performance standards for antimicrobial susceptibility testing,fifteenth informational supplement S.CLSI,2005,M 100-S15 44-50.
17、 Berger-Bachi B,Rohrer S.Factors influencing methicillin resistance in staphylococcus.Arch Microbiol,2002,178(3):165-171.
18、 Predari SC,Ligozzi M,Fontana R.Genotypic identification of methicillin-resistant cogulase-negative staphylococci by polymerase chain reaction.Antimicrob Agents Chemotherary.1991,35(12):2568-2573.
19、 Hoffmann E,Stech J,Webster RG,et al.Universalprimer set for the full-length amplification of all influenza aviruses.J Arch Virol,2001,146(12):2275.
20、 Nishi J,Miyanohara H,Nakajima T,et al.Molecular typing of the methicillin resistance determinant(mec) of clinical strains of staphylococcus based on mec hypervariable region length polymorphisms.Lab Clin Med,1995,126(1):29-35.
21、 Schwarz S,Kehrenberg C,Ojo KK,et al.staphylococcus sciuri gene erm(33),encoding inducible resistance to macrolides,lincosamides,and streptogramin B antibiotics,is a product of recombination between erm(C) and erm(A).Antimicrob Agents Chemotherapy,2002,46(2):3621-3623.
22、孙保良,李之光,张敏.葡萄球菌对大环内酯类的耐药研究.中华检验医学杂志,2006,29(5):459.
23、 Patel MV,De Souza NJ,Gupte SV,et al.Antistaphylococcal Activity of WCK 771,a tricyclic fluoroquinolone,in animal infection models.Antimicrob Agents Chemotherapy,2004,48(12):4754-4751.
24、何礼贤.耐药革兰阳性球菌的医院流行病学及感染控制.国外医药抗生素分册,2003,24(3):103-107.
25、 Archer GL,Molecular epidemiology of multiresistant staphylococcus[J].J Antimcrob Agents Chemother,1988,21(supplc):133.
26、张青,雷呈祥,赵旭.表皮葡萄球菌生物膜形成分子机制的研究进展.微生物学报,2003,43(5):681-685.
27、 Vuong C,Voyich J M,Fischer ER,et al.Polysaccharide intercellular adhesin (PIA) protects staphylococcus epidermidis against major components of the human innate immune system.Cell Microbiol,2004,6(3):269-275.
28、李敏,高谦,王建平等.表皮葡萄球菌附属基因调节因子对生物膜形成的调节作用.中华检验医学杂志,2005,28(11):1172-1177.
29、 Yao Y,Vuong C,Kocianova S,et al.Characterization of the staphylococcus epidermidis accessory-gene regulator reponse:quorum-sensing regulation of resistance to human innate host defense.J Infect Dis,2006,193(6):841-848.
30、李家泰,Allan JW,扬敏.中国细菌耐药监测研究[J].中华医学杂志,2001,81(1):8-15.
31、刑铭友,田德英,刘莉娜.2004-2005同济医院表皮葡萄球菌耐药性分析.华中科技大学学报(医学版),2007,36(5):677-680.
32、赵旭,张婴元.表皮葡萄球菌感染防治.中国抗感染化疗杂志,2003,3(5):318-320.
1、刑铭友,田德英,刘莉娜.2004-2005年同济医院表皮葡萄球菌耐药性分析.华中科技大学学报(医学版),2007,36(5):677-680.
2、赖燕菲,吴瑶,盛吉芳等.256例表皮葡萄球菌医院感染分析.浙江大学学报(医学版),2000,29(5):223-226.
3、 Versalovi CJ,Koeuth T,Lupski J R.Distribution of repetitive DNA sequences in eubecteria and application to fingerprinting of bacterial genomes[J].Nucleic Acids Res,1991,19(24):6823-6831.
4、 Silbert S,Hollis R J.Evaluation of three molecular techniques for nonfermetative Gram-negative bacill.Infect Control Hosp Epidemiol,2004,25(10):847-851.
5、 Gillings M,Holley M.Repetitive element PCR fingerprinting(rep-PCR) using enterobacterial repetitive intergenic consensus(ERIC) primers is not necessarily directed at ERIC elements[J].Letters in Applied Microbiology,1997,25(1):17-21.
6、陈迎春,曹又方,赵立平.大肠杆菌MG1655菌株ERIC-PCR图谱主带序列组成分析[J].微生物学通报,2002,29(6):28-32.
7、 Dalla-Costa L M,Irino K,Rodrigues J,et al.Characterisation of diarrhoeagenic Escherichia coli clones by ribotyping and ERIC-PCR[J].Journal of Medical Microbiology,1998,47(3):227-234.
8、黄革,董婷,侯铁英等.重复序列引物聚合酶链反应追踪金黄色葡萄球菌所致医院感染.中国感染与化疗杂志,2006,6(4):260-262.
9、 Debast SB,Meis JFGM,Melchers WJ,et al.Use of interrepeat PCR fingerprinting to investigate an Acinetobacter baumannii out-break in an intensive care unit.Scan J Infect Dis,1996,28(2):577-581.
10、 Szewcjyk EM,Piltrowski A,Rojalska M.Predominant staphylococci in the intensive care unit of a pediatric hospital[J].Hosp Infect,2000,45(2):145-154.
11、李胜利,张婴元,吴菊芳等.葡萄球菌医院感染的调查研究[J].中华医 院感染学杂志,1998,8(1):65-67.
12、熊自忠,李涛,徐元宏.临床分离耐甲氧西林表皮葡萄球菌的耐药性监测.中国医师杂志,2005,7(2):262-263.
13、 Vuong C,Otto M.Staphylococcus epidermidis infections.Microbes Infect,2002,4(4):481-489.
14、唐开洪,宋鹏,凌月明等.临床常见病原菌分布及耐药趋势分析.实用医技杂志,2003,10(6):563-564.
15、何礼贤.耐药革兰氏阳性球菌的医院流行病学及感染控制.国外医药抗生素分册,2003,24(3):103-107.
16、王海燕,梁慧,崔云龙等.重复片段PCR检测葡萄球菌DNA指纹在医院感染中的分析.中华医院感染学杂志,2003,13(6):514-516.
17、李雅卿,黄卫红.耐甲氧西林金黄色葡萄球菌医院感染分析[J].中华医院感染杂志,2002,12(2):151-152.
18、 Hulton CS,Higgins CF,Shap PM.ERIC sequences:a novel family of repetitive elements in the genomes of Escherchia coli,Salminella typhimurium and other entrobacteria.Mol Microbiol,1991,5(4):825-834.
19、 Jersek B,Tcherneva E,Rijpens N,et al.Repetitive element sequence-based PCR for species and strain discrimination in the genus Listeria.Lett Appl Microbiol,1996,23(1):55-60.
20、李艳琴,申泉,刘彬彬等.番茄内生菌分离及其ERIC-PCR指纹图谱分析.微生物学通报,2003,30(5):89-93.
21、 De Bruijin F J.Use of repetitive(repetitive extragenic palindromic and enterobacterial repetitive intergeneic consensus) sequences and the polymerase chain reaction to fingerprint the genomes of Rhizobium meliloti iaolates and other soil bacteria.Appl Environ Microbiol,1992,58(7):2180-2187.
22、 Di Giovanni GD,Watrud LS,Seidler RJ,et al.Fingerprinting of mixed bacterial strains and BIOLOG grem-negative(GN) substrate communities by enterobacterial repetitive intergenic consensus sequence- PCR (ERIC-PCR).Curr Microbiol,1999,38(4):217-223.
23、高平平,赵立平.微生物群落结构探针杂交评价不同培养基从活性污泥分离优势菌群的能力[J].微生物学报,2003,43(2):264-270.
24、李艳琴,孙永艳,崔丽芳等.ERIC-PCR在人工混合菌体系中的检出灵敏度.微生物学通报,2004,31(4):61-64.
25、李犹平,倪学勤.ERIC-PCR技术特点及其应用.安徽农业科学,2007,35(18):5358-5360.
26、晏群,陈丽华,刘文恩等.分子生物学分型技术ERIC-PCR的建立.实用医学杂志,2003,19(6):688-689.
1、 erformance standards for antimicrobial susceptibility testing;fifteenth informational supplement S.CLSI,2005,M 100-S 15 44-50.
2、 Vuong C,Otto M.staphylococcus epidermidis infections.Microbes Infect,2002,4(4):481-489.
3、唐开洪,宋鹏,凌月明等.临床常见病原菌分布及耐药趋势分析.实用医技杂志,2003,10(6):563-564.
4、 熊自忠,李涛,徐元宏.临床分离耐甲氧西林表皮葡萄球菌的耐药性监测.中国医师杂志,2005,7(2):262-263.
5、 何礼贤.耐药革兰氏阳性球菌的医院流行病学及感染控制.国外医药抗生素分册,2003,24(3):103-107.
6、 陈智鸿,吕晓菊,范昕建等.金黄色葡萄球菌耐药性监测.中国抗感染化疗杂志,2003,3(1):13-15.
7、 赖燕菲,吴瑶,盛吉芳等.256例表皮葡萄球菌医院感染分析.浙江大学学报(医学版),2000,29(5):223-226.
8、 刑铭友,田德英,刘莉娜.2004-2005年同济医院表皮葡萄球菌耐药性分析.华中科技大学学报(医学版),2007,36(5):677-680.
9、 朱德妹,汪复,张婴元.2004年上海地区细菌耐药性监测.中国抗感染化疗杂志,2005,5(4):195-200.
10、余红,李强.四年中临床分离的表皮葡萄球菌耐药性分析.实用全科医学,2005,3(4):366-367.
11、王箭,罗军.表皮葡萄球菌所致医院感染的分布及耐药趋势.徐州医学院学报,2005,25(4):338-340.
12、周桂华,尹有宽.导管相关性感染.国外医学内科学分册,1997,24(6):235-237.
13、 Barie PS.Antibiotic-resistant gram-positive cocci:implications for surgical practice.World J Surg,1998,22(2):118-126.
14、 Rupp ME,Archer GL.Coagulase-negtive staphylococci:pathogen associated with medical progress.Clin Infect Dis,1994,19(2):231-243.
15、 Lim D,Strynadka NC.Structural basis for the beta lactam resistance of PBP2α from methicillin-resistant staphylococcus aureus.Nat Struct Biol,2002,9(11):870-876.
16、 Pagliero E,Cgesnel L,Hopkins J,et al.Biochemical characterization of strephylococcus pneumonia penicillin-binding protein 2b and its implication in beta-lactam resistance.Antimicrob Agents Chemotheary,2004,48(5):1848-1855.
17、陈智鸿,吕晓菊,范昕建等.金黄色葡萄球菌耐药性监测.中国抗感染化疗杂志,2003,3(1):13-15.
18、 Schierholz JM.Beuth J.Implant infections:a heaven for opportunistic becteria [J].J Hosp Infect,2001,49(2):87-93.
19、 Vuong C,Voyich J M,Fischer ER,et al.Polysaccharide intercellular adhesin (PIA) protects staphylococcus epidermidis against major components of the human innate immune sustem.Cell Microbiol,2004,6(3):269-275.
20、江娟,孙景勇,欧元祝等.医源性表皮葡萄球菌ica操纵子转录水平对生物膜表型的影响.上海医学,2006,29(1):40-44.
21、李敏,高谦,王建平等.表皮葡萄球菌附属基因调节因子对生物膜形成的调节作用.中华检验医学杂志,2005,28(11):1172-1177.
22、 Yao Y,Vuong C,Kocianova S,et al.Characterization of the staphylococcus epidermidis accessory-gene regulator response:quorum-sensing regulation of resistance to human innate host defense.J Infect Dis,2006,193(6):841-848.