| Co-Investigator(Kenkyū-buntansha) |
TOMITA Haruyoshi Gunma University, School of Medicine, Assistant Professor, 医学部, 講師 (70282390)
FUJIMOTO Shuhei Gunma University, School of Medicine, Assistant Professor, 医学部, 講師 (90241869)
TANIMOTO Koichi Gunma University, School of Medicine, Associate Professor, 医学部, 助教授 (40188389)
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| Budget Amount *help |
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 2003: ¥4,700,000 (Direct Cost: ¥4,700,000)
Fiscal Year 2002: ¥9,400,000 (Direct Cost: ¥9,400,000)
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| Research Abstract |
(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
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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
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