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* Antibiotic resistance: PCG, penicillinase positive; DMPPC, methicillin and other Ĉ-lactams; EM, macrolides and lincomycins; KM, producing a 3'-phosphotransferase; GM, producing a dual enzyme having 6'-acethyltransferase and 2"-phosphotransferase; TOB, producing a 4', 4"-adenylyltransferase. GM: gentamicin, TOB: tobramycin, PCG: benzylpenicillin, DMPPC: methicillin, EM : erythromycin, KM: kanamycin

MCIPC: cloxacillin, DMPPC: methicillin, CPM cefpiramide, CMZ: cefmetazole, CEZ: cefazolin, CZX: ceftizoxime, IPM: imipenem, CZON: cefzonam, FMOX: flomoxef sodium Fig. 6. Induction of PBP-2' by Ĉ-lactams in Staphylococcus aureus TK388 strains (GMr-MRSA)

7) UBUKATA K, YAMASHITA N, KONNO M : Occurrence of a Ĉ- lactam - inducible penicillin - binding protein in methicillin-resistant staphylococci. Antimicrob Agents Chemother 27 : 851 ` 857, 1985 in Staphylococcus aureus. J Bacteriol 167: 975 11) SONG M D, WACIII M, Doi M, ISMINO F, MATSUHASIII M : Evolution of an inducible penicillin-target protein in methicillin-resistant Staphylococcus aureus by gene fusion. FEBS Lett 221 : 167 ` 171, 1987 13) KoNo M, S ASATSU M, O' HARA K, Siiiomi Y, H AYASAKA T:Mechanism of resistance to some cephalosporins in Staphylococcus aureus, Antimicrob Agents Chemother 23 : 938 `940, 1983 17) UBUKATA K, YAMASHITA N, GOTOH A, KONNO M: Purification and characterization of aminoglycoside -modifying enzymes from Staphylococcus aureus and Staphylococcus epidermidis, Antimicrob Agents Chemother 25: 754 `759, 1984 19) OUCHTERLONY O : Antigen-antibody reaction in gels. Acta Pathol Microbiol Scand 26: 507 20) BRADFORD M M: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye 8) UTSUI Y, YOKOTA T : Role of an altered penicillinbinding protein in methicillin- and cephemresistant Staphylococcus aureus. Antimicrob Agents Chemother 28 : 397 403, 1985 9) Rossi L, TONIN E, CHENG Y R, FONTANA R: Regulation of penicillin-binding protein activity : description of a methicillin- inducible penicillinbinding protein in Staphylococcus aureus. Antimicrob Agents Chemother 27: 828 831, 1985 10) MATSUHASHI M, SONG M D, ISHINO F, WACHI M, DOI M, I NOUE M, UBUKATA K, YAMASHITA N, KONNO M: Molecular cloning of the gene of a penicillin-binding protein supposed to cause high resistance to Ĉ-lactam antibiotics binding. Anal Biochem 72 : 248 `254, 1976 21) UBUKATA K, NONOGUCHI R, MATSUHASHI M, K ONNO M Expression and inducibility in Staphylococcus aureus of the meca gene, which encodes a methicillin-resistant S. aureus -specific penici-llin-binding protein. J. Bacteriol 171: 2882 `2885, 1989 22) MANIATIS T, FRITSCH E F, S AMBROOK j M olecular cloning : a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N. Y. 23) JEvoNs M P : "Celbenin"-resistant Staphylococci. Brit Med J 1 : 124 `125, 1961

25) BARRETT F F, MCGEHEE R I, FINLAND M: Methicillin-resistant Staphylococcus aureus at Boston City Hospital. N Engi J Med 279 : 441 26) PARKER M T, HE:WITT J H : Methicillin resistance in Staphylococcus aureus. Lancet 1 : 800 ` 804, 1970 27) K LIMEK J J, MAI4Slk F J, BARTI.ETIT R C, B, SHEP UINTILIANI R : Clinical, epidemiological and bacteriologic observations of an outbreak of methicillin - resistant Staphylococcus aureus at a large community hospital. Am J Med 61 : 340 `345, 1976 28) CROSSLEY K, LOESCH D, LANDESMAN B, MEAD K, CHERN M, STRATE R: An outbreak of infections caused by strains of Staphylococcus aureus resistant to methicillin and aminoglycosides. I.Clinical Studies J Infect Dis 139 : 273 ` 279, 1979 29) SAPICO J F, MONTGOMERIE J Z, CANAWATI H N, AEILTS G D: Methicillin - resistant Staphylococcus aureus. Am J Med Sic 281 : 101 ` 109, 1981 30) S CHAEFLER S, J ONES D, PERRY W, RUVINSKAYA L, BARODET Y, MAYA E, WILSON M E : Emergence of gentamicin and methicillinresistant Staphylococcus aureus strains in New York City Hospitals. J. Clin. Microbiol. 13 :754 `759, 1981 31) COLLINS J K, M ADER J T, K ELLY M T: Resistance of methicillin - resistant Staphylococcus aureus to third-generation cephalosorins. J Infect Dis 147 : 591 `596, 1983 35) UHIJKATA K, NoN0GUCOL R, MATSWIASJU M, SONG M D, KONNO M Restriction maps of the regions coding for methicillin and tobramycin resistance on chromosomal DNA in methicillinresistant staphylococci. Antimicrob Agents Chemother. 33 : 1624 `1626, 1989 36) HARTMAN B J,TOMASZ A: Low-affinity penicillinbinding protein associated with Ĉ-lactam resistance in Staphylococcus aureus. J. Bacteriol 158: 513 516, 1984 38) M URAKAMI K, N OMCRkA K, Doi M, YosimA T: Production of low-affinity penicillinbinding protein by low- and high-resistance groups of methicillin - resistant Staphylococcus aureus. Antimicrob Agents Chemother. 31: 1307 `1311, 1987

METHICILLIN -RESISTANT STAPHYLOCOCCI ISOLATED FROM BLOOD CULTURE EPIDEMIOLOGIC CHARACTERISTICS AND INDUCTION OF PENICILLIN - BINDING PROTEIN-2' BY Ĉ- LACTAMS RITSUKO NONOGUCHI Department of Clinical Pathology, Teikyo University School of Medicine, (Director : Prof. KONNO M), 2-11-1, Kaga, Itabashi-ku, Tokyo 173, Japan 1. Methicillin-resistant Staphylococcus aureus (MRSA) was investigated for its isolation frequency from clinical materials from 1980 to 1988. The rapid increase of MRSA from the beginning of 1980 coincided with the introduction into the market of new antipseudomonal penicillins and second- and third-generation cephems. 2. The change in the rate of isolates obtained from blood culture in the aforementioned period showed that S. aureus, of gram-positive cocci, was evidently increasing, whereas the isolation rate of gram-negative bacilli changed little or, in some species, even showed a slight decrease. 3. MRSA strains isolated from blood culture during the past five years were epidemiologically classifiable into three major groups: gentamicin-resistant (GM) MRSA, group 1, was typed using bacteriophage group I and coagulase IV, which produced enterotoxin A or B. Chromosomal DNA encoding the meca gene was a 4.0 kb Hind III fragment ; tobramycin-resistant (TOBr) MRSA, group 2, was typed using phage group III and coagulase II, which produced toxin shock syndrome toxin 1 (TSST-1) alone or TSST-1 and enterotoxin C. The DNA encoding the meca gene was a 4.3 kb Hind III fragment ; GMr+TOBr-MRSA, group 3, was similar to group 2 epidemiologically. These MRSA strains indicated rapid changes from group 1 to group 2, then to group 3. 4. Induction of penicillin-binding protein (PBP)-2' in MRSA of these three groups by or flomoxef was low, whereas in the other MRSA groups, PBP-2' induction was prevalent with all the compounds tested. 5. From these results, it appears that MRSA are acquiring more adaptability to new environments along with the change in the use of antibiotics.

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