Analysis of virulence factors of enterococcus, that are adhesion, bacteriocin and vancomycin resistance.

肠球菌毒力因子粘附性、细菌素及万古霉素耐药性分析。

• 批准号: 14370089
• 负责人: IKE Yasuyoshi
• 金额: $ 9.02万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370089/
• 关键词: enterococcus; genetics; E.faeciunt; E.faecalis; adhesin; bacteriocin; plasmid; vancomycin resistance; Enterococcus faecalis; Enterococcus faecium; バンコマイシン耐性腸球菌(VRE); ゲンタマイシン耐性プラスミド; 接合伝達性プラスミド; 接合凝集

(1)Genetic Analysis of the Bacteriocin 32 Determinant encoded on the Plasmid Isolated from Vancomycin Resistant Enterococcus faecium Clinical Isolate.The ability of E.faecalis clinical isolates to adhere to immobilized extracellular matrixes (ECMs) coating the walls of microtiter plates was examined by microscope. The ECMs consisted of fibronectin, laminin, collagen I, II, IV, V, fibrinogen and lactoferrin. The tissue specific adhesive strains AS11, AS12, AS14, AS15, HT11 and HT12, which show highly efficient adherence to human bladder carcinoma T24 cells and human bladder epithelial cells, showed strong adherence to each of the fibronectin, laminin, collagen I, II, IV, and V EC Ms, at levels greater than 10^4 cells per mm^2 of well surface coated by ECM.(2)Tissue Specific Adherent Enterococcus faecium Strains that show Highly Efficient Adhesion to Human Bladder Carcinoma T24 Cells also adhere to Extracellular matrix Proteins.Of the 636 vancomycin-resistant Enterococcus faecium (VRE) i … More solates, two hundred seventy-seven (44%) of the strains were bacteriocinogenic, and one hundred ninety-three (30.3%) of the strains showed bacteriocin activity against E.faecium and E.hirae. The bacteriocinogenic VRE200 strain harbored highly efficiently conjugative gentamicin resistance plasmid pG200 (60kb), and other plasmid p200B encoded 58 amino acids, which was the bacteriocin determinant.(3)Possible connection between a widely disseminated conjugative gentamicin resistance (pMG1-like) plasmid and the emergence of vancomycin resistance in Enterococcus faecium.A total of 640 vancomycin-resistant Enterococcus faecium (VRE) isolates from the Medical School Hospital of the University of Michigan, Ann Arbor, were used in this study. Of the 611 and 29 were VanA and VanB VRE, respectively, based on PCR analysis. The gentamicin resistance of each of 261 (53%) of the 492 gentamicin-resistant strains was transferred to E.faecium at a frequency of about 10^<-5> to 10^<-6> per donor cell in broth mating. The conjugative gentamicin resistance plasmids were identified and were classified into five types (A through B) with respect to their EcoRI restriction profiles. The EcoRI or NdeI restriction fragments of each type of plasmids hybridized to the conjugative gentamicin resistance plasmid pMG1 (65.1kb), which was originally isolated from an E.faecium clinical isolate, and transfer efficiently in broth mating. Less

（1）万古霉素耐药屎肠球菌临床分离株质粒上细菌素32决定簇的遗传分析通过显微镜检查粪肠球菌临床分离株粘附于包被在微量滴定板壁上的固定化细胞外基质（ECM）的能力。ECM由纤连蛋白、层粘连蛋白、胶原蛋白I、II、IV、V、纤维蛋白原和乳铁蛋白组成。组织特异性粘附菌株AS 11、AS 12、AS 14、AS 15、HT 11和HT 12显示出对人膀胱癌T24细胞和人膀胱上皮细胞的高效粘附，显示出对纤连蛋白、层粘连蛋白、胶原蛋白I、II、IV和V EC Ms中的每一种的强粘附，其水平大于10^4个细胞/mm^2的由ECM包被的孔表面。（2）对人膀胱癌T24细胞表现出高效粘附的组织特异性粘附屎肠球菌菌株也粘附于细胞外基质蛋白。 ...更多信息 分离物中，277株（44%）菌株是细菌素原性的，193株（30.3%）菌株显示出对屎肠球菌和平肠球菌的细菌素活性。产细菌素的VRE 200菌株含有高效接合型庆大霉素抗性质粒pG 200（60 kb），另一质粒p200 B编码58个氨基酸，是细菌素的决定簇。（3）广泛分布的接合性庆大霉素耐药（pMG 1-like）质粒与屎肠球菌万古霉素耐药的可能关系本研究共收集了640株来自密歇根大学医学院医院的耐万古霉素屎肠球菌（VRE）。根据PCR分析，611和29分别为VanA和VanB VRE。492株庆大霉素耐药菌株中的261株（53%）的庆大霉素耐药性在肉汤交配中以约10 - 10 μ g/供体细胞的频率转移到屎肠球菌<-5><-6>中。鉴定了接合性庆大霉素抗性质粒，并根据其EcoRI限制性图谱将其分为5种类型（A至B）。每种质粒的EcoRI或NdeI限制性片段与接合性庆大霉素抗性质粒pMG 1（65.1kb）杂交，并在肉汤交配中有效转移，所述接合性庆大霉素抗性质粒pMG 1最初是从屎肠球菌临床分离株分离的。少

项目成果

Tomita H, Pierson C, Lim SK, Clewell DB, Ike Y.: "Possible connection between a widely disseminated conjugative gentamicin resistance (pMG1-like) plasmid and the emergence of vancomycin resistance in Enterococcus faecium."J Clin Microbiol.. 40-9. 3326-333

Tomita H、Pierson C、Lim SK、Clewell DB、Ike Y.：“广泛传播的接合庆大霉素抗性（pMG1 样）质粒与屎肠球菌中万古霉素抗性的出现之间可能存在联系。”J Clin Microbiol.. 40-

Francia MV, Ike Y, Clewell DB. et al.: "Completion of the nucleotide sequence of the Enterococcus faecalis conjugative virulence plasmid pAD1 and identification of a second transfer origin"Plasmid. 46. 117-127 (2001)

弗兰西亚 MV、艾克 Y、克莱威尔 DB。

Tomita H, Tanimoto K, Hayakawa S, Morinaga K, Ezaki K, Oshima H, Ike Y.: "Highly conjugative pMG1-like plasmids carrying Tn1546-like transposons that encode vancomycin resistance in Enterococcus faecium."J Bacteriol.. 185・23. 7024-7028 (2003)

Tomita H、Tanimoto K、Hayakawa S、Morinaga K、Ezaki K、Oshima H、Ike Y.：“高度接合的 pMG1 样质粒携带 Tn1546 样转座子，编码屎肠球菌中的万古霉素抗性。”J Bacteriol.. 185・23 .7024-7028 (2003)

Ike Y, Arakawa Y, Ma X, Tatewaki K, Nagasawa M, Tomita H, Tanimoto K, Fujimoto S.: "A nationwide survey shows that vancomycin heterogeneously resistant MRSA and vancomycin intermediately resistant MRSA are not disseminated throughout Japanese hospitals"Jo

Ike Y、Arakawa Y、Ma X、Tatewaki K、Nagasawa M、Tomita H、Tanimoto K、Fujimoto S.：“一项全国性调查显示，万古霉素异质耐药性 MRSA 和万古霉素中度耐药性 MRSA 并未在日本医院中传播”Jo

Tomita H, Pierson C, Lim SK, Clewell DB, Ike Y.: "Possible connection between a widely disseminated conjugative gentamicin resistance (pMG1-like) plasmid and the emergence of vancomycin resistance in Enterococcus faecium."J Clin Microbiol.. 40・9. 3326-333

Tomita H、Pierson C、Lim SK、Clewell DB、Ike Y.：“广泛传播的接合庆大霉素抗性（pMG1 样）质粒与屎肠球菌中万古霉素抗性的出现之间可能存在联系。”J Clin Microbiol.. 40。 9.3326-333