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Restriction Endonuclease Analysis of Plasmid DNA From Methicillin-Resistant Staphylococcus aureus: Clinical Application Over a Three-Year Period

  • Antoni Trilla (a1) (a2), Mary D. Nettleman (a2) (a3), Richard J. Hollis (a4), Mary Fredrickson (a3), Richard P. Wenzel (a2) and Michael A. Pfaller (a5)...



To investigate trends in plasmid types of methicillin-resistant Staphylococcus aureus over a 3-year period and determine the clinical impact of plasmid typing.


Restriction endonuclease analysis of plasmid DNA (REAP) was used to type 270 clinical isolates.


A Veterans Affairs Medical Center.


Forty-four different REAP types were identified and, of these, 23 appeared only once. Types designed as V1 and V2 were more commonly found in nosocomial cases than in community-acquired cases (p<.05); whereas the K and D types were more common in isolates from community-acquired cases (<c.05). In 30 (11%) isolates, no plasmid was detected. When combined with epidemiological data, REAP typing revealed 4 small outbreaks that would have been missed using traditional epidemiological methods alone. In large outbreaks involving 10 or more cases, REAP typing data revealed unsuspected patterns of transmission.


Multiple plasmid types were present in this endemic setting. Restriction endonuclease analysis of plasmid DNA was a practical and valuable adjunct to traditional epidemiological methods.


Corresponding author

University of Iowa Hospitals , C41GH, Iowa City, Iowa 52242


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1.Preheim, LC, Rimland, D, Bittner, MJ. Methicillin-resistant Stufihylococcus aureus in Veterans Administration Medical Centers. Infect Control. 1987;8:191194.
2.Storch, GA, Radcliff, JL, Meyer, PL, Hinrichs, JH. Methicillin-resistant Staphylococcus aureas in a nursing home. Infect Control. 1987;8:2429.
3.Hsu, CC, Macaluso, CP, Special, L, Hubble, RH. High rate of Methicillin-resistance of Staphylococcus aureus isolated from hospitalized nursing home *patients. Arch Intern Med. 1988;148:569570.
4.Thomas, CJ, Bridge, J, Waterman, S, Vogt, J, Kilman, L, Hancock, G. Transmission and control of methicillin-resistant Staphylococcus uureus in a skilled nursing facility. Infect Control Hasp Epidemiol. 1989;10:106110.
5, Haley, RW, Hightower, AW, Khabbaz, RFet al. The emergence of methicillin-resistant Staphylococcus uureus infections in United States hospitals. Ann Intern Med. 1982;97:297308.
6.Standiiord, HC. Methicillin-resistant Staphylococcus uureus infections: it's time to get tough. Infect Control. 1987;5:187189.
7.Crossley, K, Loesch, D, Iandesman, B, Mead, K, Chern, M, Strate, RAn outbreak of infections caused by strains of Staphylococcus uureus resistant to methicillin and aminoglycosides. I. Clinical studies. J Infect Dis. 1979;139:273279.
8.Giamorellou, H, Papetropoulou, M, Daikos, GK. Methicillin-resistant Staphylococcus uureus infections during 19781979: Clinical and bacteriologic observations. J Antimicrob Chemother. 1981;7:649655.
9.Thompson, RL, Cabezudo, I, Wenzel, RF! Epidemiology of nosocomial infections caused by methicillin-resistant Staphylococcus uureus. Ann Intern Med. 1982;97:309317.
10.Boyce, JM. Increasing prevalence of methicillin-resistant Stuphylococcus uureus in the United States. Infect Control Hosp Epidemiol. 1990;11:639642.
11.Jones, RN, Barry, AL, Gardiner, RV, Packer, RRThe prevalence of staphylococcal resistance penicillinase-resistant penicillins. A retrospective and prospective national surveillance trial of isolates from 40 medical centers. Diugn Microbial Infect Dis. 1989;12:385394.
12.Boyce, JM, White, RL, Spruill, EY. Impact of methicillin-resistant Staphylococcus aureus on the incidence of nosocomial staphylococcal infections.] Infect Dis. 1983;148:763.
13.Locksley, RM, Cohen, ML, Quinn, TC, et al. Multiply antibiotic-resistant Staphylococcus aureus: introduction, transmission and evolution of nosocomial infection. Ann Intern Med. 1982;97:317324.
14.Nettleman, MD, Trilla, A, Fredrickson, M, Pfaller, M. Assigning responsibility: using feedback to achieve sustained control of methicillin-resistant Staphylococcus uureus. Am J Med. 1991;91 (suppl 3B):228S232S.
15.Wenzel, RP, Nettleman, MD, Jones, RN, Pfaller, MA. Methicillin-resistant Staphylococcus uureus: implications for the 1990s and effective control measures. Am J Med. 1991;91 (suppl 3B):221S227S.
16.Peacock, JE, Marsik, FJ, Wenzel, RP. Methicillin-resistant Staphylococcus aureus: introduction and spread within a hospital. Ann Intern Med. 1980;93:526532
17.Schaefler, S, Jones, D, Perry, W, et al. Methicillin-resistant Staphylococcus aureus in New York City hospitals: inter-hospital spread of resistant strains of type 88. J Clin Microbial. 1984;20:536538.
18.Saravolatz, LD, Pohlod, DJ, Arking, LM. Community-acquired methicillin-resistant Staphylococcus aureus infections: a new source of nosocomial outbreaks. Ann Intern Med. 1982;97:325329.
19.Wakefield, DS, Pfaller, MA, Massanari, RM, Hammons, GT. Variation in methicillin-resistant Staphylococcus aureus occurrence by geographic location and hospital characteristics. Infect Control. 1987;8:151157.
20.Archer, GL, Mayhall, CG. Comparison of epidemiological markers used in the investigation of an outbreak of methicillin-resistant Staphylococcus aureus infections. J Clin Microbial. 1983;18:395399.
21.Licitra, CM, Brooks, RG, Terry, PM, Shaw, KJ, Hare, RS. Use of plasmid analysis and determination of aminoglycoside modify ing enzymes to characterize isolates from an outbreak of methicillin-resistant Staphylococcus aureus. J Clin Microbial. 1989;27:25352538.
22.Pfaller, MA, Wakefield, DS, Hollis, R, Fredrickson, M, Evans, E, Massanari, RM. The clinical microbiology laboratory as an aid in infection control: the application of molecular techniques in epidemiologic studies of methicillin-resistant Staphylococcus aureus. Diagn Microbial Infect Dis. 1991;14:209217.
23.Mayer, LW. Use of plasmid profiles in epidemiologic surveillance of disease outbreaks and in tracing the transmission of antibiotic resistance. Clin Microbial Rev. 1988;1:228243.
24.Nahaie, MR, Goodfellow, M, Harwood, CR. A rapid screening procedure for staphylococcal plasmids. J Microbial Method. 1984;2:7381.
25.Hartstein, AI, Morthland, VH, Eng, S, Archer, GL, Schoenknect, FD, Rashad, AL. Restriction enzyme analysis of plasmid DNA and bacteriophage typing of paired Staphylococcus aureus blood culture results. J Clin Microbiol. 1989;27:18741879.
26.Pfaller, MA. Hollis, RI. Use of nlasmid profiles and restriction endonuclease analysis of plasmid DNA as epidemiologic and diagnostic tools in the clinical microbiology laboratory. Clin Microbial Newslet. 1989;11:137141.
27.Zuccarelli, AJ, Roy, I, Harding, GP, Couperus, JJ. Diversity and stability of restriction enzyme profiles of plasmid DNA from methicillii-resistant Staphylococcus aureus. J Clin Microbial. 1990;28:97102.
28.Lyon, BR, May, JW, Skurray, RA. Analysis of plasmids in nosocomial strains of multiple antibiotic-resistant Staphylococcus aureus. Antimicrob Agents Chemother. 1983;23:817826.
29.Rhinehart, E, Shlaes, DM, Keys, TF, et al. Nosocomial clonal dissemination of methicillin-resistant Staphylococcus aureus. Arch Intern Med. 1987;147:521524.
30.Mulligan, ME, Arbeit, RD. Epidemiologic and clinical utility of typing systems for differentiating among strains of methicillin-resistant Staphylococcus aureus. Infect Control Hosp Epidemiol. 1991;12:2028.
31.Fournier, JM, Bouvet, A, Boutonnier, A, et al. Predominance of capsular polysaccharide type 5 among oxacillin-resistant Staphylococcus aureus. J Clin Microbial. 1987;25:19321933.
32.Mulligan, ME, Kwok, RY, Citron, DM, John, JF Jr, Smith, PB. Immunoblots, antimicrobial resistance, and bacteriophage typing of oxacillin-resistant Staphylococcus aureus. J Clin Microbial. 1988;26:23952401.
33.Jordens, JZ, Hall, LM. Characterisation of methicillin-resistant Staphylococcus aureus isolates by restriction endonuclease digestion of chromosomal DNA. J Med Microbial. 1988;27:117123.
34.Branger, C, Goullet, P, Boutonnier, A, Foumier, JM. Correlation between esterase electrophoretic types and capsular polysaccharide types 5 and 8 among methicillin-susceptible and methicillin-resistant strains of Staphylococcus aureus. J Clin Microbial. 1990;28:150151.
35.Bumie, JP, Matthews, RC, Lee, W, Murdoch, D. A comparison of immunoblot and DNA restriction patterns in characterising methicillin-resistant isolates of Staphylococcus aureus. J Med Microbial. 1989;29:255261.
36.Reboli, AC, John, JF Jr, Levkoff, AH. Epidemic methicillin-gentamicin-resistant Staphylococcus aureus in a neonatal intensive care unit. J Dis Child. 1989;143:3439.
37.Gamer, JS, Jarvis, WR, Emori, TG, Horan, TC, Hughes, JM. CDC definitions for nosocomial infections. Am J Infect Control. 1988;16:128140.
38.Broderick, A, Mori, M, Nettleman, MD, Streed, SA, Wenzel, RP. Nosocomial infections: validation of surveillance and computer modeling to identify patients at risk. Am J Epidemiol. 1990;131:734742.
39.Thornsberry, C, McDougal, LK. Successful use of broth microdilution in susceptibility tests for methicillin-resistant (heteroresistam) staphylococci. J Clin Microbial. 1983;18:10841091.
40.National Committee for Clinical Laboratory Standards. Approved Standard M2-A3. Performance Standards for Antimicrobial Disk Susceptibility Tests. Villanova, Pa: National Committee for Clinical Laboratory Standards; 1984.
41.McDougal, LK, Thomsberry, C. New recommendations for disk diffusion antimicrobial susceptibility tests for methicillin-resistant (heteroresistant) staphylococci. J Clin Microbial. 1984;19:482488.
42.Opal, SM, Mayer, KH, Stenberg, MJ, et al. Frequent acquisition of multiple strains of methicillin-resistant Staphylococcus aureus by healthcare workers in an endemic hospital environment. Infect Control Hosp Epidemiol. 1990;11:479485.
43.Reboli, AC, John, JF, Platt, CG, Cantey, JR. Methicillin-resistant Staphylococcus aureus outbreak at a Veterans' Affairs Medical Center: importance of carriage of the organism by hospital nersonnel. Infect Control Hosp Epidemiol. 1990;11:291295.
44.Muder, RR, Brennen, C, Wagener, MM, et al. Methicillin-resistant staphylococcal colonization and infection in a long-term care facility. Ann Intern Med. 1991;114:107112.

Restriction Endonuclease Analysis of Plasmid DNA From Methicillin-Resistant Staphylococcus aureus: Clinical Application Over a Three-Year Period

  • Antoni Trilla (a1) (a2), Mary D. Nettleman (a2) (a3), Richard J. Hollis (a4), Mary Fredrickson (a3), Richard P. Wenzel (a2) and Michael A. Pfaller (a5)...


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