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Application of Molecular Techniques to the Study of Nosocomial Infections Caused by Enterococci

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Molecular Bacteriology

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 15))

Abstract

Enterococci are components of the normal bowel flora of humans and other animals, and have traditionally been considered to be of relatively low virulence in healthy individuals. However, they are increasingly important nosocomial pathogens and have been cited as the leading organism isolated from hospital-acquired infections, and the third leading cause of nosocomial bacteremia in the United States in a recent National Nosocomial Infection Surveillance (NNIS) system report of the Centers for Disease Control (1). The increase in enterococcal infections has been associated with the emergence of resistance to multiple antibiotics, in particular resistance to B-lactams, high-level aminoglycoside resistance, and resistance to glycopeptides. Concern that antibiotic resistance will continue to spread and will increasingly render conventional antimicrobial chemotherapy inadequate for serious enterococcal infections has stimulated interest in methods to improve the diagnosis and epi-demiologic investigation of infections caused by enterococci

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References

  1. Schaberg, DR., Culver, D.H, and Gaynes, R P(1991) Major trends in the microbial etiology of nosocomial Infection Am J Med 91, 72S–75S

    PubMed  CAS  Google Scholar 

  2. Teixeira, L M, Facklam, R R, Steigerwalt, A G, Pigott, N E, Merquior, V L C, and Brenner, D.J (1995) Correlation between phenotypical characteristics and DNA relatedness within Enterococcus faecium strains J Clin Microbial 33 1520–1523

    CAS  Google Scholar 

  3. Williamson, R Gutmann, L, Horaud, T, Delbos, F, and Acar, J (1986) Use of pemcillin-binding proteins for the identification of enterococci J Gen Microbiol 132, 1929–1937.

    CAS  Google Scholar 

  4. Vincent, S, Minkler, P., Bincziewski, B., Etter, L, and Shlaes, D M(1992) Vancomycin resistance in Enterococcus gallnarum Antimicrob Agents Chemother 36, 1392–1399

    CAS  Google Scholar 

  5. Donabedian, S., Chow, J. W, Shlaes, D M., Green, M., and Zervos, M J (1995) DNA hybridization and contour-clamped homogeneous electric field electrophoresis for identification of enterococci to the species level J Clin Microbiol 33, 141–145

    PubMed  CAS  Google Scholar 

  6. Facklam, R R, and Sahm, D A (1995) Enterococcus, in Manual of Clinical Microbiology sixth ed (Murray, P R, Baron, E J, Pfaller, M A, Tenover, F.C, and Yolken, R H, eds ), American Society of Microbiology, Washington,DC, 308–314

    Google Scholar 

  7. Pitcher, D G, Saunders, N A, and Owen, R J. (1989) Rapid extraction of bacterial genomic DNA with guanidium thiocyanate. Lett App Microbiol 8, 151–156

    CAS  Google Scholar 

  8. Wilson, K(1994) Preparation of genomic DNA from bacteria.2 4.1–2.4 2.Greene, Brooklyn, NY

    Google Scholar 

  9. McClelland, M, Petersen, C., and Welsh, J (1992) Length polymorphisms in tRNA intergenic spacers detected by using the polymerase chain reaction can distinguish streptococcal strains and species J Clin Microbiol 30, 1499–1504

    PubMed  CAS  Google Scholar 

  10. Hynes, W. L, Ferretti, J. J., Gilmore, M. S, and Segarra, R E(1992) PCR amplification of streptococcal DNA using crude cell lysates FEMS Microbiol Lett 94, 139–142

    CAS  Google Scholar 

  11. Klare, I, Heier, H, Claus, H, Reissbrodt, R, and Witte, W (1995) vanA-mediated high-level glycopeptide resistance inEnterococcus faecium from animal husbandry FEMS Microbiol Lett 125, 165–172

    PubMed  CAS  Google Scholar 

  12. Dutka-Malen, S., Leclercq, R, Coutant, V, Duval, J, and Courvalin, P(1990) Phenotypic and genotypic heterogeneity of glycopeptide resistance determinants in gram-positive bacteria. Antimicrob Agents Chemother 34, 1875–1879

    PubMed  CAS  Google Scholar 

  13. Cocconcelli, P S, Porro, D, Galandini, S., and Senim, L (1995) Development of RAPD protocol for typing of strains of lactic bacteria and enterococci. Lett App Microbiol 21, 376–379

    CAS  Google Scholar 

  14. Tomayko, J F., Zscheck, K. K, Singh, K V., and Murray, B. E (1996) Comparison of the β-lactamase gene cluster in clonally distinct strains of Enterococcus faeclis Antimicrob Agents Chemother 40, 1170–1174

    CAS  Google Scholar 

  15. Sambrook, J, Fritsch, E F., and Maniatis, T. (1989) Molecular cloning A Laboratory Manual 2nd Ed CSHL

    Google Scholar 

  16. Daly, J A, Clifton, N L., Seskm, K. C, and Manford Gooch, W. (1991) Use of rapid, nonradioactive DNA probes in culture confirmation tests to detect Streptococcus agalactiae, Haemophilus influenzae, and Enterococcus spp from pediatric patients with significant infections. J Clin. Microbiol 29, 80–82

    PubMed  CAS  Google Scholar 

  17. Davis, T E, and Fuller, D. D. (1991) Direct identification of bacterial isolates in blood cultures by using a DNA probe. J Clin. Microbiol 29, 2193–2196.

    PubMed  CAS  Google Scholar 

  18. Betzl, D, Ludwig, W, and Scheleifer, K. H. (1990) Identification of lactococci and enterococci by colony hybridization with 23S rRNA-targeted oligonucleotides probes. Appl Environ Microbiol 56, 2927–2929

    PubMed  CAS  Google Scholar 

  19. Beimfohr, C., Krause, A, Amann, R, Ludwig, W, and Schleifer, K H. (1993) In situ identification of lactococci, enterococci and streptococci.Sys Appl Microbiol 16, 450–456

    Google Scholar 

  20. Costa, Y., Galimandd, M., Leclercq, R, Duval, J, and Courvalm, P. (1993) Characterization of the chromosomal aac(6’)-Ii gene specific for Enterococcus faecium Antimicrob. Agents Chemother 37, 1896–1903

    CAS  Google Scholar 

  21. Leclercq, R, Dutka-Malen, S, Duval, J., and Courvalin, P (1992) Vancomycin resistance gene vanC is specific to Enterococcus gallinarum. Antimirob. Agents Chemother.36, 2005–2008

    CAS  Google Scholar 

  22. Navarro, F, and Courvalin, P (1994) Analysis of genes encoding D-alanine-D-alanine ligase-related enzymes inEnterococcus casseliflavus and Enterococcus flavescens. Antimicrob Agents Chemother.38, 1788–1793

    CAS  Google Scholar 

  23. Coque, M T., and Murray, BE (1995) Identification of Enterococcus faecalis strains by DNA hybridization and pulsed-field electrophoresis gel J Clan Microbiol 33, 3368–3369.

    CAS  Google Scholar 

  24. Dutka-Malen, S., Evers, S, and Courvalin, P. (1995) Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci by PCR. J Clin. Microbiol 33, 24–27.

    PubMed  CAS  Google Scholar 

  25. Clark, N C, Cooksey, RC, Hill, BC., Swenson, J. M., and Tenover, FC (1993) Characterization of glycopeptide-resistant enterococci from U.S. hospitals.Annmicrob Agents Chemother 37, 231l–2317

    Google Scholar 

  26. DutkaMalen, S, Evers, S., and Courvalin, P. (1995) Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci by PCR (Published erratum) J Clin Microbiol 33, 1434

    CAS  Google Scholar 

  27. Cartwright, C. P., Stock, F., Fahle, G A., and Gill, V. J (1995) Comparison of pigment production and motility tests with PCR for reliable identification of intrinsically vancomycin-resistant enterococci. J. Clin Microbiol.33, 1931–1933

    PubMed  CAS  Google Scholar 

  28. Lowe, A M., Lambert, P. A., and Smith, A. W (1995) Cloning of an Enterococcus faecalis endocarditis antigen: homology with adhesin from some oral streptococci.Infect Immun.63, 703–706

    PubMed  Google Scholar 

  29. Xu, Y, Jiang, L., Murray, B. E., and Wemstock, G. M (1996)Enterococcus faecalis antigens in human infections. in press.

    Google Scholar 

  30. Korten, V., Huang, W.M., and Murray, B. E. (1994) Analysis by PCR and direct DNA sequencing of gyrA mutations associated with fluoroqumolone resistance in Enterococcus faecalis Antimicrob Agents Chemother 38, 2091–2094.

    CAS  Google Scholar 

  31. Chu, C., Kariyama, R, Daneo-Moore, L., and Shockman, G. (1992) Cloning and sequence analysis of the muramidase-2 gene from Enterococcus hirae J.Bactenol 174, 1619–1625.

    CAS  Google Scholar 

  32. Fontana, R, Cermi, R., Longoni, P, Grossato, A., and Canepari, P (1983) ldentification of a streptococcal penicillin-binding protein that reacts slowly with penicilln J Bacteriol 156, 1343–1350.

    Google Scholar 

  33. Grayson, M. L., Eliopoulos, G M, Wennersten, C. B., Ruoff, K L., Klimm, K, Sapico, F. L, Bayer, A. S., and Moellering, R C. (1991) Comparison of Enterococcusraffinosus with Enterococcus avium on the basis of penicillin susceptibility,penicillin-binding protein analysis, and high-level aminoglycoside reststance. Antimicrob Agents Chemother 35, 1408–1412

    PubMed  CAS  Google Scholar 

  34. Collins, M. D, Facklam, R. R, Farrow, J.A.E, and Williamsom, R. (1989) Enterococcus raffinosus ssp. nov., Enterococcus solitarius ssp. nov and Enterococcus pseudoavium ssp. nov FEMSM microbiol Lett 57, 283–288

    Google Scholar 

  35. Merquior, V.L.C, Peralta, J.M, Faklam, R R, and Teixeira, L M. (1994) Analysis of whole-cell protein profiles as a tool for characterization of Enterococcus species Curr Microbiol 28, 149–153.

    CAS  Google Scholar 

  36. Murray, B. E (1990) The life and times of the enterococcus Clin Mxrobiol Rev. 3, 46–65

    CAS  Google Scholar 

  37. Coudron, P. E, Mayhall, C G, Facklam, R R., Spadors, A.C, Lamb, V A, Lybrand, M. R, and Dalton, H. P (1984)Streptococcus faecium outbreak in a neonatal intensive care unit.J. Clin Microbiol 20, 1044–1048.

    PubMed  CAS  Google Scholar 

  38. Kuhnen, E., Richter, F., Rtchter, K., and Andries, L (1988) Establishment of a typying system for group D streptococci, Zentralbl Bakteriol Parasitenkd Infektionskr Hyg Abt I Orig Reihe A 267, 322–330

    CAS  Google Scholar 

  39. Tomayko, J F, and Murray, B.E (1995) Analysts of Enterococcus faecalis isolates from intercontinental sources by multilocus enzyme electrophoresis and pulsed-field gel electrophoresis. J Clinn Microbiol 33 2903–2907

    CAS  Google Scholar 

  40. Kuhn, I Burman, L G., Hoeggman, S., Tullus, K., and Murray, BE (1995) Biochemical fingerprinting compared with ribotyping and pulsed-field gel electrophoresis ofDNA for epidemilogical typing of enterococci J Clin Microbiol 33, 2812–2817

    PubMed  CAS  Google Scholar 

  41. Klare, I, Heier, H., Claus, H., Bohme, G, Marin, S., Seltmann, G., Harenbeck, R, Antanassova, V, and Witte, W.(1995)Enterococcus faecium strains with vanA mediated high level resistance isolated from animal foodstuffs and fecal samples of humans in the community Microbial Drug Res.1 265–272.

    CAS  Google Scholar 

  42. Huycke, M. M., Spiegel, C A.,and Gilmore, M S. (1991) Bacteremia caused by hemolytic, high-level gentamicin-resistant Enterococcus faecalis. Antimicrob Agents Chemother 35, 1626–1634.

    CAS  Google Scholar 

  43. Luginbuhl, L. M, Rotbart, H.A, Facklam, R R., Roe, M H., and Elliot, J A (1987) Neonatal enterococcal sepsis: case-control study and description of an outbreak. PediatrInfect Dis.6, 1022–1030

    CAS  Google Scholar 

  44. Chirurgi, V. A, Oster, S E., Goldberg, A.A, Zervos, M. J., and McCabe, R E.(1991) Ampicillin-resistant Enterococcus raffinosus in an acute-care hospital: case-control study and antimicrobial susceptibilities. J Clin Microbiol 29, 2663–2665

    PubMed  CAS  Google Scholar 

  45. Boyce, J.M., Opal, S.M., Potter-Bynoe, G., LaForge, R G., Zervos, M. J., Furtado, G., Victor, G, and Medeiros, A. A. (1992) Emergence and nosocomial transmission of ampicillin-resistant enterococci. Antimicrob Agents Chemother. 36, 1032–1039.

    PubMed  CAS  Google Scholar 

  46. Donabedian, S.M., Chow, J.W., Boyce, J.M., McCabe, R.E., Markowitz, S M., Coudron, P E, Kuntza, A, Pierson, C L., and Zervos, M J (1992) Molecular typing of ampicillin resistant, non-βlactamase producing Enterococcus faecium from diverse geographical areas J Clin Microbiol 30, 2757–2761

    PubMed  CAS  Google Scholar 

  47. Woodford, N, Momson, D, Cookson, B, and George, R C. (1993) Companson of high-level gentamicin-resistant itEnterococcus fuecium isolates from different continents Antimicrob Agents Chemother 31, 681–684

    Google Scholar 

  48. Antalek, M.D., Mylotte, J M, Lesse, A J, and Sellick, J A (1995) Clinical and molecular epidemiology ofEnterococcus faecalis bacteremia, with special reference to strains with high level resistance to gentamicin CID 20, 103–109.

    CAS  Google Scholar 

  49. Zervos, M.J, Kauffman, C A, Therasse, P.M, Bergman, A G, Mikesell, T.S,and Schaberg, D R. (1987) Nosocomial Infection by gentamicin-resistant Streptococcus faecalis an epidemiologic study. Ann Intern. Med 106, 687–691

    PubMed  CAS  Google Scholar 

  50. Thal, L A, Chow, J.W, Patterson, J.E, Perri, M B, Donabedian, S, Clewell, D B, and Zervos, M J (1993) Molecular characterlzatlon of highly reslstant Enterococcus faecalis isolates lackmg high level streptomycm resistance Antimicrob Agents Chemother 37, 134–137

    PubMed  CAS  Google Scholar 

  51. Patterson, J. E., Masecar, B L, Kauffman, C. A., Schaberg, D R, Hlerholzer, W J, Jr., and Zervos, M J (1988) Gentamlcin resistance plasmlds of enterococcl from diverse geographic areas are heterogeneous J Infect Dis 158, 212–216

    PubMed  CAS  Google Scholar 

  52. Rhinehart, E., Smith, N E, Wennersten, C., Gorss, E, Freeman, J., Eliopoulos,G.M, Moellering, R. C., Jr, and Goldmann, D. A. (1990) Rapid dissemination of β-lactamase-producing, ainoglycoside-resistant Enterococcus faecalis among patients and staff on an infant-toddler surgical ward N Engl J Med 323, 1814–1818

    PubMed  CAS  Google Scholar 

  53. Boyce, J M., Opal, S M, Chow, J.W, Zervos, M.J, Potter-Bynoe, G., Sherman, C.B, Romulo, M C., Fortna, S., and Medeiros, A A (1994) Outbreak of multidrug resistant Enterococcus faecium with transferable vanB class vancomycin resistance J Clin Microbiol 32, 1148–1153.

    PubMed  CAS  Google Scholar 

  54. Murray, B.E, Lopardo, H A., Rubegho, E A., Frosolono, M., and Singh, K V (1992) Intrahospital spread of a single gentamicin-resistant, β-lactamase-producing strain ofEnterococcus faecalis in Argentina. Antimicrob Agents Chemother 36, 230–232.

    PubMed  CAS  Google Scholar 

  55. Patterson, J E., Wanger, A., Zscheck, K K, Zervos, M.J, and Murray, B.E.(1990) Molecular epidemiology of β-lactamase-producing enterococci Antimicrob. Agents Chemother 34, 302–305

    PubMed  CAS  Google Scholar 

  56. Bingen, E H., Denamur, E., Lambert-Zechovsky, N.Y, and Elion, J (1991)Evidence for the genetic unrelatedness of nosocomial vancomycin-resistant Enterococcus faecium strains in a pediatric hospital. J Clin Microbiol 29,1888–1892

    PubMed  CAS  Google Scholar 

  57. Hall, L M., Duke, B, Gurney, M., and Williams, R. (1992) Typing of Enterococcus species by DNA restriction fragment analysis. J Clin Microbiol 30, 915–919

    PubMed  CAS  Google Scholar 

  58. Lacoux, P.A., Jordens, J Z, Fenton, C.M., Guiney, M., and Pennington, T H. (1992) Characterization of enterococcal isolates by restriction enzyme analysis of genomic DNA. Epidemiol Infect 109, 69–80

    PubMed  CAS  Google Scholar 

  59. Singh, K V, and Murray, B.E (1994) Revised estimates of enterococcal chromosomal sizes DNA Cell Biol 13, 1145–1146

    PubMed  CAS  Google Scholar 

  60. Murray, B E., Singh, K.V, Heath, J. D, Sharma, B R., and Weinstock, G M. (1990) Comparison of genomic DNAs of different enterococcal isolates using restriction endonucleases with infrequent recognition sites.J. Clin Microbiol 28, 2059–2063

    PubMed  CAS  Google Scholar 

  61. Vander Auwera, P, Pensart, N., Korten, V, Murray, B E., and Leclercq, R (1996) Incidence of oral glycopeptides on the fecal flora of human volunteers: selection of highly glycopeptide resistant enterococci. J infect Dis 173, 1129–1136

    PubMed  Google Scholar 

  62. Plessis, P., Lamy, T., Donnio, P Y., Autuly, F., Grulois, I., LePrise, P Y, and Avril, J L (1996) Epidemiologic analysis of glycopeptide-resistant Enterococcus strains in neutropenic patients receiving prolonged vancomycin adminstration Eur J Clin Microbiol Infect Dis 14, 959–963

    Google Scholar 

  63. Christie, C, Hammond, J, Reising, S, and Patterson, J E. (1994) Clinical and molecular epidemiology of enterococcal bacteremia in a pediatric teaching hospital J Pediatrics 125, 392–399

    CAS  Google Scholar 

  64. Morris, T, Brecher, S M, Fitzsimmons, D., Durbin, A, Arbeit, R D., and Maslow, J N. (1995) A pseudoepidemic due to laboratory contamination deciphered by molecular analysis Infect Con. Hosp Epidemiol 16, 82–87.

    CAS  Google Scholar 

  65. Miranda, A.G., Singh, K V, and Murray, B E (1991) DNA fingerprinting of Enterococcus faecium by pulsed-field gel electrophoresis may be a useful epidemiologic tool. J Clin Microbiol 29, 2752–2757

    PubMed  CAS  Google Scholar 

  66. Murray, B.E, Singh, K V., Markowitz, S.M., Lopardo, H. A, Patterson, J E, Zervos, M.J., Rubeglio, E., Eliopoulos, G. M., Rice, L. B., Goldstein, F.W, Jenkins, S.G., Caputo, G M., Nasnass, R, Moore, L S., Wong, E.S, and Weinstock, G. (1991) Evidence for clonal spread of a single strain of plactamase-producing Enterococcus faecalis to six hospitals in five states.J Infect Dis. 163, 780–785

    PubMed  CAS  Google Scholar 

  67. Green, M., Barbadora, K., Donabedian, S., and Zervos, M.J (1995) Comparison of field inversion gel electrophoresis with contour clamped homogeneous electric field electrophoresis as a typing method for Enterococcus faecium J Clin. Microbiol 33, 1554–1557.

    CAS  Google Scholar 

  68. Boyle, J F., Soumakis, S. A., Rendo, A., Herrington, J.A, Gianarkis, D G, Thurberg, B.E., and Painter, B.G. (1993) Epidemiologic analysis and genotypic characterization of a nosocomial outbreak of vancomycin-resistant enterococci. J Clin Microbiol 31, 1280–1285.

    PubMed  CAS  Google Scholar 

  69. Tenover, F C, Arbeit, R.D, Goering, R.V, Mickelsen, P. A, Murray, B E, Persing, DH, and Suaminathan, B (1995) cnInterpreting chromosomal DNA restriction patterns produced by pulsed field gel electrophoresis. criteria for bacterial strain typing J Clin Microbiol. 33, 2233–2239

    PubMed  CAS  Google Scholar 

  70. Quiniliani, R., Jr, and Courvalin, P. (1994) Conjugal transfer of the vancomycin resistance determinant vanB between enterococci involves the movement of large genetic elements from chromosome to chromosome.FEMS Microbiol Lett. 119, 359–364.

    Google Scholar 

  71. .Seetulsingh, P.S., Tomayko, J. F., Coudron, P.E., Markowitz, S.M, Skinner, C, Singh, K V., and Murray, B E. (1996) Chromosomal DNA restriction endonuclease digestion patterns of beta-lactamase producing Enterococcus faecalis isolated from a single hospital over a 7-year period. J Clin Microbiol 34, 1892–1896.

    PubMed  CAS  Google Scholar 

  72. Jordens, J Z., Bates, J., and Griffiths, D. T. (1994) Faecal carriage and nosocomial spread of vancomycin-resistant Enterococcus faecium J Antimicrob Chemother 34,515–528.

    CAS  Google Scholar 

  73. Woodford, N., Morrison, D., Johnson, A P, Briant, V., George, R.C, and Cookson, B (1993) Application of DNA probes for rRNA and vanA genes to investigation of a nosocomial cluster of vancomycin-resistant enterococci.J Clin. Microbiol 31, 653–658.

    PubMed  CAS  Google Scholar 

  74. Gordillo, M.E, Singh, K V, and Murray, B E (1993) Comparison of ribotyping and pulsed-field gel electrophoresis for subespecies differentiation of Enterococcus faecalis J Clin Microbiol 31, 1570–1574.

    CAS  Google Scholar 

  75. Bates, J., Jordens, J Z, and Griffiths, D.T. (1994) Farm animals as a putative reservoir for vancomycin-resistant enterococcal infection in man. J Antimicrob Chemother 34, 507–516

    PubMed  CAS  Google Scholar 

  76. Thorisdottir, A.S, Canas, L.L, Marshall, S H., Green, M, Zervos, M.J, and Glorgio, C. (1994) IS6770, an insertion-like sequence useful for determining the clonal relationship of clinical enterococcal isolates J Infect Dis 170, 1539–1548.

    PubMed  CAS  Google Scholar 

  77. vanBelkum, A. (1994) DNA fingerprinting of medically important microorganisms by use of PCR. Clin. Microbiol Rev 7, 174–184

    PubMed  Google Scholar 

  78. Barbier, N., Saulnier, P, Chachaty, E., Dumontier, S., and Andremont, A. (1996) Random amplified polymorphic DNA typing versus pulsed-field gel electrophoresis for epidemlological typing of vancomycin resistant enterococci.J Clin Microbiol.34, 1096–1099.

    PubMed  CAS  Google Scholar 

  79. Kostman, J.R., Alden, M B., Mair, M., Edlind, T. D., LiPuma, J J., and Stull,T. L (1995) A universal approach to bacterial molecular epidemiology by polymerase chain reaction ribotyping.J Infect. Dis 171, 204–208

    PubMed  CAS  Google Scholar 

  80. vanAsselt, G. J, Vliegenthart, J.S, Petit, P.L.C., vande Klundert, J.A.M,and Mouton, R.P (1992) High-level aminoglycoside resistance among enterococci and group A streptococci. J. Antimicrob Chemother 30, 651–659.

    PubMed  Google Scholar 

  81. Weems, J.J, Lowrance, J.H., Baddour, L.M., and Simpson, W.A. (1989) Molecular epidemiology of nosocomial, multiply aminoglycoside resistant Enterococcus faecalis J. Antimicrob. Chemother 24, 121–130

    CAS  Google Scholar 

  82. Ounissi, H., Derlot, E., Carher, C., and Courvalin, P (1990) Gene homogeneity for aminoglycoside-modifying enzymes in gram-positive cocci. Antimicrob.Agents Chemother 34, 2164–2168

    PubMed  CAS  Google Scholar 

  83. Quintiliani, R, Jr, Evers, S, and Courvalin, P (1993) The vanB gene confers various levels of self-transferable resistance to vancomycin in enterococci J Infect Dis 167, 1220–1223

    PubMed  CAS  Google Scholar 

  84. Hayden, M.K, Trenholme, G M, Schultz, J E, and Sahm, D F. (1993) In viva development of teicoplanin resistance in a VanB Enterococcus faecium isolate J Infect Dis 167, 1224–1227

    PubMed  CAS  Google Scholar 

  85. Green, M, Binczewski, B, Pasculle, A W, Edmund, M, Barbadora, K, Kusne, S., and Shlaes, D (1993) Constitutively vancomycin-resistant Enterococcus faecium resistant to synergistic β-lactam combinations Antimicrob Agents Chemother 37, 1238–1242

    PubMed  CAS  Google Scholar 

  86. Swenson, J.M, Clark, N.C, Ferraro, M J, Sahm, D F, Doem, G, Pfaller, M A, Reller, L B, Weinstein, M P, Zabransky, R J, and Tenover, F C (1994) Development of a standardized screening method for detection of vancomycin-resistant enterococci J Clin Microbiol 32, 1700–1704

    PubMed  CAS  Google Scholar 

  87. Sahm, D F, and Gilmore, M S (1994) Transferability and genetic relatedness of high level gentamicin resistance among enterococci Antimicrob Agents Chemother 38, 1194–1196

    PubMed  CAS  Google Scholar 

  88. Leclercq, R, Dutka-Malen, S, Brisson-Noel, A, molinas, C, Derlot, E, Arthur, M, Duval, J, and Courvalin, P (1992) Resistance of enterococci to aminoglycosides and glycopeptides CID 15, 495–501

    CAS  Google Scholar 

  89. Leclercq, R, Bismuth, R, and Duval, J (1992) New high content disks for determination of high level aminoglycoside resistance in clinical isolates of Enterococcus faecalis Eur J Clin Microbiol Infect Dis 11, 356–360

    CAS  Google Scholar 

  90. Sahm, D F, Boonlayangoor, S, Iwen, P C., Baade, J L, and Woods, G L (1991) Factors influencing determination of high level amioglycoside resistance in Enterococcus faecalis J Clin Microbiol 29, 1934–1939.

    CAS  Google Scholar 

  91. Bantar, C E., Micucci, M, Femandez Camgia, L, Smayevski, J, and Bianchini, H M (1993) Synergy characterization for Enterococcus faecalis strains displaying moderately high level gentamicin and streptomycin resistance J Clin Microbiol 31, 1921–1923

    PubMed  CAS  Google Scholar 

  92. Moellering, R C, Jr., Murray, B.E, Schoenbaum, S.C., Adler, J,and Wennersten, C.B (1980) A novel mechanism of resistance to penicillin-gen-tamicin synergism in Streptococcus faecalis J Infect. Dis 141, 81–86

    CAS  Google Scholar 

  93. Wennersten, C. B., and Moellering, R.C, Jr. (1980) Mechanism of resistance to penicillin-aminoglycoside synergism in Streptococcus faecium, in Current Chemotherapy and Infectious Diseases Vol 1, (Nelson, J and Grassi, C, eds), American Society for Microbiology, Washington, DC, 710–712

    Google Scholar 

  94. Thal, L.A, Chow, J W, Mahayni, R, Bonilla, H, Perri, MB., Donabedian, S A, Silverman, J, Taber, S, and Zervos, M J (1995) Characterization of antimicrobial resistance in enterococci of animal origin Antimicrob Agents Chemother 39, 2112–2115

    PubMed  CAS  Google Scholar 

  95. Coque, T M., Arduino, R C, and Murray, B E. (1995) High-level resistance to aminoglycosides: Comparison of community and nosocomial fecal isolates of enterococci. Clin Infect Dis.20, 1048–1051

    PubMed  CAS  Google Scholar 

  96. Kaufhold, A, Podbielski, A, Horaud, T., and Ferrreri, P (1992) Identical genes confer high level of resitance to gentamicin uponEnterococcus faecalis, Enterococcus faectium, and Streptococcus agalactiae.Antimicrob Agents Chemother 36, 1215–1218.

    PubMed  CAS  Google Scholar 

  97. Patterson, J.E., and Zervos, J M (1990) Hugh-level gentamicin in Enterococcus. microbiology, genetic basis, and epidemiology. Rev. Infect Dis.12, 644–652

    PubMed  CAS  Google Scholar 

  98. Tenover, F.C, Swenson, J.M, O’Hara, C M, and Stocker, S A. (1995) Ability of commercial and reference antimicrobial susceptibility testing methods to detect vancomycin resistance in enterococci J Clin. Microbiol 33, 1524–1527

    PubMed  CAS  Google Scholar 

  99. Tenover, F.C, Tokars, J., Swenson, J, Paul, S, Spitalny, K, and Jarvis, W (1993) Ability of clinical laboratories to detect antimicrobial agent resistant enterococci. J Clin. Microbiol 31, 1695–1699

    PubMed  CAS  Google Scholar 

  100. Snell, J. J.S, Brown, D.F J, Perry, S. F, and George, R. (1993) Antimicrobial susceptibility testing of enterococci. results of a survey conducted by the United Kingdom National External Quality Assesment Scheme for Microbiology J Antimicrob Chemother 32, 401–411.

    PubMed  CAS  Google Scholar 

  101. Woodford, N, Johnson, A. P, Morrison, D., and Speller, D C E (1995) Current perspectives on glycopeptide resistance Clin Microbiol Rev 8, 585–615

    PubMed  CAS  Google Scholar 

  102. Handwerger, S., Skoble, J., Discotto, L F, and Pucci, M J (1995) Heterogeneity of the vanA gene cluster in clinical isolates of enterococci from Northeastern United States.Antimicrob Agents Chemother.39, 362–368.

    PubMed  CAS  Google Scholar 

  103. Aarestrup, F M., Ahrens, P, Madsen, M., Pallesen, L V, Poulsen, R L., and Westh, H (1996) Glycopeptide susceptibility among Danish Enterococcus faectium and Enterococcus faecalis isolates of animal and human origin and PCR identification of the genes within the VanA cluster Antimicrob Agents Chemother 40, 1938–1940

    PubMed  CAS  Google Scholar 

  104. Coque, T.M, Tomayko, J F., Ricke, S C., Okhyusen, P C., and Murray, B E (1996) Vancomycin Resistant Enterococci from Nosocomial, Community and Animal Sources in the United States. submitted

    Google Scholar 

  105. Jett, B.D, Huycke, MM., and Gilmore, M S. (1994) Virulence in enterococci Clin Microbiol Rev.7, 462–478.

    PubMed  CAS  Google Scholar 

  106. Coque, T M, Patterson, J.E, Steckelberg, J M., and Murray, B E. (1995) Incidence of hemolysin, gelatinase and aggregative substance among enterococci isolated from patients with endocarditis and other infections and from feces of hospitalized and community-based persons, J Infect Dis 171, 1223–1229.

    PubMed  CAS  Google Scholar 

  107. Huycke, M.M., and Gilmore, M.S. (1995) Frequency of aggregation substance and cytolysin genes among enterococcal endocarditis isolates.Plasmid 34, 152–156

    PubMed  CAS  Google Scholar 

  108. Aitchison, E J., Lambert, P A., Smith, E G., and Farrell, I. D (1987) Serodiagnoses of Streptococcus faecalis endocarditis by immunoblotting of surface protein antigens J,Clin Microbiol 25, 211–215

    CAS  Google Scholar 

  109. Lambert, P.A, Shorrock, P.J., Aitchison, E.J., Domingue, P A. G, Power, M E, and Costerton, J W (1990) Effect of in vivo growth conditions upon expression of surface protein antigens in Enterococcus faecalis FEMS Microbiol Immun 64, 51–54

    Google Scholar 

  110. Shorrock, P J., Lambert, P A, Aitchison, E J., Smith, E G, Farrell, I.D., and Gutschik, E. (1990) Serological response in Enterococcus faecalis endocarditis determined by enzyme-linked immunosorbent assay.J Clin Microbiol 28, 195–200

    PubMed  CAS  Google Scholar 

  111. Li, X., Weinstock,G M., and Murray, B E (1995) Generation of auxotrophs mutants in Enterococcus faecalis J Bacteriol 177, 6866–6873.

    PubMed  CAS  Google Scholar 

  112. Anderson, D G, and McKay, L L (1983) Simple and rapid method for isolating large plasmid DNA from lactic streptococci Appl Environ Microbiol 46

    Google Scholar 

  113. Weaver, K.E., and Clewell, D.B (1988) Regulation of the pAD1 sex pheromone response in Enterococcus faecalis: construction and characterization of lacZ transcriptional fusions in a key control region of the plasmid J Bacteriol 170, 4343–4352

    PubMed  CAS  Google Scholar 

  114. Wanger, A.R., and Murray, B.E. (1990) Comparison of enterococcal and staphylococcal beta-lactamase plasmids J Infect Dis 161, 54–58

    PubMed  CAS  Google Scholar 

  115. Dutka-Malen, S, Molinas, C., Arthur, M, and Courvalin, P. (1990) The VANA glycopeptide resistance protein is related to D-alanyl-D-alanine ligase cell wall biosynthesis enzymes Mol Gen Genet 224, 366–372.

    Google Scholar 

  116. Evers, S, Sahm, D F, and Courvalin, P. (1993) The vanB gene of vancomycin-resistant Enterococcus faecalis V583 is structurally related to genes encoding D-AlaD-Ala ligases and glycopeptide-resistance proteins VanA and VanC Gene 124, 143–144

    PubMed  CAS  Google Scholar 

  117. Gold, H.S., Unal, S, Cercenado, E, Thauvin-Eliopoulos, C., Eliopoulos, G. M., Wennersten, C B., and Moellering, R C., Jr.(1993) A gene conferring resistance to vancomycin but not teicoplanin in isolates of Enterococcus faecalis and Enterococcus faeclium demonstrates homology with vanB, vanA and vanC genes of enterococci Antimicrob Agents Chemother 37, 1604–1609

    PubMed  CAS  Google Scholar 

  118. Dutka-Malen, S., Molinas, C., Arthur, M., and Courvalin, P. (1992) Sequence of the vanC gene of Enterococcus gallinarum BM4174 encoding a D-alanine D-alanine ligase-related protein necessary for vancomycin resistance Gene 112, 53–58.

    PubMed  CAS  Google Scholar 

  119. Trieu-Cuot, P, and Courvalin, P (1983) Nucleotide sequence of the Streptococcus faecalis plasmid gene encoding the 3′5′-aminoglycoside phosphotransferase type III Gene 23, 331–341

    PubMed  CAS  Google Scholar 

  120. Ferretti, J.J, Gilmore, K.S., and Courvalin, P(1986) Nucleotide sequence analysis of the gene specifying the bifunctional 6′-aminoglycoside acetyltransferase 2′-aminolgycoside phosphotransferase enzyme in Streptococcus faecalis and identification and cloning of gene regions specifying the two activities. J Bacteriol 167, 631–638

    PubMed  CAS  Google Scholar 

  121. Galli, D., and Worth, R (1991) Comparative analysis of Enterococcus faecalis sex pheromone plasmids identifies a single homologous DNA region which codes for aggregation substance. J Bacteriol.173, 3029–3033

    PubMed  CAS  Google Scholar 

  122. Segarra, R.A., Booth, M C., Morales, D.A., Huycke, M.M., and Gilmore, M. S (1991) Molecular characterization of theEnterococcus faecalis cytolysin activator Infect Immun 59, 1239–1246

    PubMed  CAS  Google Scholar 

  123. Gilmore, M S., Segarra, R.A., Booth, M.C., Bogie, C P, Hall, L R, and Clewell, D.B. (1994) Genetic structure of the Enterococcus faecalis plasmid pADl-encoded cytolytic toxin system and its relationship to lantibiotic determinnts J Bacteriol 176, 7335–7344

    PubMed  CAS  Google Scholar 

  124. vander Klundert, J.A M., and Vlegenthart, J.S. (1993).PCR detection of genes coding for aminoglycoside-modifying enzymes, in Diagnostic Molecular Microbiology Principles and Applications (Persing, D.H., Smith, T F., Tenover, F C., and Whiote, T.J., eds.), American Society for Microbiology, Washington, DC, 547–552

    Google Scholar 

  125. Torres, C., Reguera, J A, Sanmartin, M.J., Perez-Draz, J C, and Baquero, F (1994) vanA-mediated vancomycin-resistant Enterococcus spp in sewage.J Antimicrob Chemother 33, 553–561.

    PubMed  CAS  Google Scholar 

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Coque, T.M., Seetulsingh, P., Singh, K.V., Murray, B.E. (1998). Application of Molecular Techniques to the Study of Nosocomial Infections Caused by Enterococci. In: Woodford, N., Johnson, A.P. (eds) Molecular Bacteriology. Methods in Molecular Medicine™, vol 15. Humana Press. https://doi.org/10.1385/0-89603-498-4:469

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