153 ῌ῎ ῏῍ Staphylococcus aureus 1)1) 1) !2)"#3) 1) $%&' 2) $%'%() 3) $%& *ICT + *, 16 4 - 12 ./ , 16 8 - 16 .+ 2002 7 - 2002 12 -01 10 2 Staphylococcus aureus 3 4 567 7 289:;< S. aureus 3/ 5 S. aureus 1=>?/ @1!A4 7 2 S. aureus 9 5 23BCDEFG *B+ II H methicillin resistant S. aureus (MRSA)/ IJ 2 29BVII HKBIII H methicillin susceptible S. aureus (MSSA) 8LM4 BVII HN3 S. aureus 9/ oxacillin (MPIPC) 1 4 8 mg/ml MIC 8 borderlineresistant S. aureus 8LM4 AA/ PCR 1O mecA penicillin binding protein2῍ (PBP2῍) 9P8LM4 Q/ 7 2 S. aureus R 20 N @!A pulsed-field gel electrophoresis (PFGE) STUV/ 20"WX#$9YZM?>3/ 289 1 2@[\/ S. aureus "WX#$ 9]^F_@`A4 5,a S. aureus K S. aureus 9/ bN1%&cO5 K3`d4 S. aureus 3O9/ efF_N@'g1() A/ efF_N1O'ghi21j*cOk+3lm4 Key words: / borderline-resistant S. aureus, PCR (polymerase chain reaction), PFGE (pulsed-field gel electrophoresis)/ nohi p3q1>?/ 1980 r,st / @1(bKZJ/ %A@ - u . methicillin-resistant Staphylococcus t&hiKZO5K<LO4 5 aureus(MRSA)9/v/1wq1xQM4yz89{ t/ &hi1B3C28LO4 |}0~<-?J/ &hi1 2Zb8LO 4 5 MRSA 9/ b-lactam 1), 2) 311A?4567 , (PBP2῍) @ &hiDE9/ FDE@G1 H1 ¡hi¢IJDEKA?/ £W/ KL nohiDE@¤¥cO5K3¦28LO4 cO5K1J.@8cO5K39?> MRSA 9nohi1JMNA/ 'ghi§Z O4 59 b-lactam 3:z1>?<6 ¨%A©ª@«A;ghi@O2cOt/ 7 ,3k+Zt/ MRSA 9 b-lactam 3b noDEPQ3¬tO4 5nohi¢IK ;@Q3?<cO 4 A?9/ MN/ %A© 3) Q/ Staphylococcus 9='Z1>?Ac STU´µ (¶693ῌ8501) ·¸¹VW 89ῌ1 $%&' TEL: 0853ῌ20ῌ2420 E-mail: smorikiºshimane-med.ac.jp ª@«cOMN&'i/ MNZk+3lmO3/ ®KAhi¢I 9i%A©ª¯°@«Ahi8LO4 ±'1>?/ b3O5K3LO4 5 b ²R19/ {|b>³ G»¼ Vol. 14 No. 3 2004. 25 154 tuvw=xy¸B=z{ ¹=º{|'=»}~' 2ῌ !(R1 OP_>/?PDx2 S. au- !"# reus !VgzMW2. kg& $%&'()*+,-. $*/ kh!"DheXY!(D. X! &04)1 232. )&456* MRSA 7 Z[21 . 89 :8;<=II ;>50$%3/ BC'+ PCR % PFGE ! -&05), 6)1 $*/$%3/?!% (R1 PCR S. aureus P!g=k #&. ;@* pulsed-field gel electro- g W. g\]2. ^; DNA %21 phoresis (PFGE) &ABC@ Tokue /_¡2 mecA " !%2+"D/*EE06)1 FG)*)*. !"D8)1 PCR ~g~k : 'H#$%&3/6* S. aureus ' ¢N=£hmkh< !"D1 Y` ED+()*C,IBC U¤M 94¥ 30 alm¦§ 55¥ 30 ! -+21 a kh¢c 72¥ 60 a! 35 ~kg(R 1 PCR . 1¨ fh!"Dbu© cF]. ª} 2002 , 7 J3/ 2002 , 12 J? BacT/ALERT %# 1,339 bp e!«[f521 3D :-.KLMN< "OP/01Q26*3 &3/. S. aureus 4ED++! =oed5e PFGE ªh= :-. BIO-RAD< ! "D. P&/?5/0]. ge!2. (R1 $S?OP/0T5U% 540 U lysozyme/lysostaphin gh. 50¥ 5 proteinase K 5. VWX 75 U&OP/0650R1 V2+. gh. ij. 25¥ 5 SmaI gh]. 1¨ bu© 10 43/ S. aureus &6*&. 3 4OP c" agarose kl] CHEF-DRIII :-. BIO-RAD< /03/Y S. aureus &6*1 OPZ[$ !"D+. b¬ 6.0 v/cm, gh? 5.3 3/ 34.9 %&3/\ S. aureus &6*. 7 4ED a. bu©c? 20.0 ?`U5bu©c21 V ++21 S. aureus 4ED+. 7] 1 8 2+. ª} ?Z93&:;,IV*/&3/ S. 21 =oed]«[f5® aureus &6*+D3<3!^21 S. aureus BacT/ALERT 3D 56!=2/0 13//0P!Q22. OP_>/? :`abC<. 1) S. au- c8d_>/?. eMfghi@/? :- reus 7 .jklmn< ,Iogp HK _> #$%&'()*+ !" /? :qABrs< /?5tu. Cvfh,Iw 7 43/6* 20 >mn% u/0Dx!EyF>ED+. z9{ ,I8=;!=2 :U 1<1 7 4o8= |. VITEK AMS :-.KLMN< ,I8 II ; 5 4 14 >. MPIPC O2+ 16 mg/ml 9{|"}~ `a :`abC< !" -¯D MIC !=2. °¤¢M±. p D89{|!(D S. aureus %21 °¦ ±. zg ±. ±,Ike±#² . NCCLS !G 7) 2*HP +p50R1 232. 4 3 3/6* 3 > &I!!"D MIC-2000 :Js< 5 clindamycin (CLDM) 0.13 mg/ml Z MIC KDxLMN (MIC) !y. oxacillin (MPIPC) 50R1 'q. vancomycin (VCM) #²+ O# MIC 3/ 2 mg/ml Z! methicillin-suscep- 50R1 8=;8=II ;& 5 4%8=VII tible S. aureus (MSSA) %2 4 mg/ml ZP! MRSA ;,I8=III ;&V*³* 1 450R1 . %21 . 'H#VQ& 3/ S. OP/03/6* S. aureus 8=;%. OP aureus &6*+D%. V>!"D+ Z[$%&3/6* S. aureus 8=; P%R,Ioe}"8 ´4?5#²+'50R1 7 4WX 4 4 9;"SOT :lza< !"DU; IVH zgrs3/\ S. aureus & !(R1 . MPIPC O2+ 0.5 mg/ml Z 6*&. µ- 3 43/ IVH zg 3/ 8 mg/ml MIC !=2'H# 4 >E rs&36*+D 3R1 . OP¶· D+ MRSA-LA a :lza< !"D+ PBP 3/ S. aureus &6* 3 4 1 >& MRSA 26 -.()*C¼ Vol. 14 No. 3 2004. G@ABud CCDE! S. aureus -.V#@ABV#6x2 1 155 CCDE!efgh2@AB[*jk 1 ' F G H I J IVHMN OPQR 4 8 0.5 4 0.13 0.13 0.13 0.13 2 ' F G 16 16 8 8 32 32 32 32 32 16 8 32 32 32 32 32 16 8 8 8 3 'FG (8/ 9) 16 'FG (8/13) 16 H I J 16 8 8 8 32 32 32 32 32 16 8 0.13 8 32 32 32 32 32 16 8 0.13 8 32 32 32 32 32 16 8 0.13 8 4 FGz% 16 FG{% 16 IVHMN 16 8 8 8 32 32 32 32 32 32 32 8 32 32 32 32 32 32 32 8 8 16 8 8 8 4 4 4 MPIPC ABPC CEZ EM CLDM OFLX MINO AMK VCM CTM CMZ CZX FMOX IPM 0.25 0.25 0.25 0.25 1 8 1 1 1 4 4 4 4 4 2 2 0.5 0.5 0.5 0.5 0.06 0.06 0.06 0.06 0.5 0.13 0.5 0.25 0.5 0.25 0.5 0.25 0.5 0.13 0.5 0.25 0.5 0.13 0.5 8 4 4 4 4 4 4 1 1 1 1 VII VII VII VII 4 8 8 4 1 2 II II 2 4 2 4 4 2 1 1 1 II II II 8 8 8 8 8 8 8 8 8 1 1 1 II II II 5 F G m 16 16 8 8 32 32 32 32 32 32 32 32 32 32 16 16 8 8 4 4 8 8 8 8 4 4 1 1 II II 6 ' F G F G IVHMN 16 16 16 16 8 8 8 8 32 32 32 32 32 32 32 32 16 16 16 16 8 8 8 8 4 4 4 4 8 8 8 8 8 8 8 8 16 32 16 8 1 2 2 1 II II II II 7 F G IVHMN 1 0.5 8 4 1 1 1 1 0.5 0.06 0.5 0.13 0.5 0.25 1 0.5 0.06 0.5 0.13 0.5 0.25 2 1 1 III III 32 32 32 32 32 32 32 32 4 1 4 2 32 32 32 32 ( mg/ml) II 2 MSSA V 1 VII 1 2) 1 S. aureus 4 mecA MPIPC 0.5 mg/ml 4 mg/ml 8 mg/ml MIC 1, 4 PCR ! PCR "#$%&'()*+ , 1 -./0 MRSA 12+34*5 1,339 bp 6789 mecA :+;<=>-. /0 MSSA ?2+34*5 @AB 1 CDE!'FG HIJ IVH KL*L5MN@A BOPQR 4 1,339 bp 6789:+; <=S T MRSA-LA UVWX YZ[ L\]^_`abc?2 IVH KL*L5M N MPIPC 0.5 mg/ml MIC > d'FG HIJ@ABOPQRCDE ! 3 efgh2Cijk MPIPC 89 MIC > 4l8 mg/ml m9 boderline-resistant ,1 1 CDE! S. aureus mecA S. aureus no 3) 7 PFGE +;$%&'()*+ , 2 4 z 7 pq 4 rZ S. aureus 17 $% % {%|}~\3DE!C9 &'()*+ stS9c u!v > z%|}{%|}CDE!G/0 !ACDE! @Z 8wxy: S. aureus PFGE $%&'()*+ 3 -.V# Vol. 14 No. 3 2004. 27 ±'²³e()´aµe¶· ¸e¹· "eº»¼" 156 ¬2 _)W6 S. aureus 7 V PFGE MPIPC /LJ borderline resistant S. aureus IVH S. aureus #$8 * PCR +J mecA )W+$ PCR MRSA ,#7 !"#$ mecA )W6 F$8 ABC MRSA-LA ῌ @ PBP2ῌ +)WF$8 MSSA ῍ %&'()*+ NCCLS ,-./01 , Louie 11) MPIPC /L57 MIC 2 MPIPC / 4 mg/ml 23 MIC +45 S. aureus 8 mg/ml mecA {( borderline resistant S. MRSA ,6 7 8 4 mg/ml 23+45 S. aureus 37 X)|xy MRSA borderline aureus 9(:; PBP < b-lactamase = resistant S. aureus +/Q7qr >?@ABC b-lactam DE<FGHI mstu+J7 K)*8B1F J73) K)*/AJ MPIPC /L X/RS[ PQ PBP2ῌ +)W57q J break point 4 mg/ml / 1 MNO 2 8 rmstuh PCR JF:+ mg/ml MIC +4 MRSA MSSA ,/ \/ MRSA )W P57 S. aureus /QGQRS57 TU V 1 #78#$ ) W 6 4 X MPIPC / L 5 7 MIC \/_ S. aureus 10 V IVH )W6 1 X 0.5 mg/ )W6 1z 7 V _2 ml 2Y Z[ 3 X )` S. aureus )W6 12) 4 8 mg/ml \ MIC _ #$ ] ] K)*/J7%&' ()* ^_`abc MIC S. epidermidis, MRSA, Staphylococcus spp. ABC MPIPC /L Staphylococcus )W6 J 0.5 mg/ml 2Y MSSA #$ PCR mecA MSSA /)WiJ7 j )W6 deIV f heterogeneous MRSA + MRSA ) W 6 V 86¡ MSSA gh PCR (+i9) j Nakatomi 70¡ /M iJ7 £¤/AJ!! _d klmnop`+&Dqrmstu ¢6 J8+ v MRSA PBP2ῌ +)W57)*+w ¥q¦{(§¨©/ª MRSA ) MPIPC /L57%&'(xy 232 X MRSA W6 J7 TU ] ] ghV /A 1z 231 Xqrmstuv( Z[ J 7 V « 4 V / IVH ¢6 1X PCR mecA {()|xy+4 MRSA )* 3 V MRSA, sm}~/qrmstuv(+i 1 V MSSA )W6 8 _"#X J710) TUghV 1 4 X1z 3 X 2)`)W6 S. aureus 1z 28 $%@&®¯° Vol. 14 No. 3 2004. @A-#.'' S. aureus KLefg>-#.?fg> II 5 14 3 157 \Gs$6D6HI"1S 3 11 MIC MIC O]^_!`a>3 MRSA N67( B8 15) !"#$ KAb %& ' " 7 20 PFGE ( ! ')*+,$-#. " ῌ ῏ 2002 c 7 2002 c 12 Q"dc/@ A S. aureus @A / MRSA 0"1& PFGE WX S. aureus 7 "/- 23&4567 20 +>$?!( KLefg /289:+ >-#.?fg>' ; 4 - <= !"# IVH "#"4567 1) 7 5 K MRSA " 2 K MSSA "1& MRSA 5 14 8ZII " MSSA 2 VII S. aureus +-#.>$?"1& ῌ"1& 7 @A"#' ' S. aureus 8Z "# S. aureus % Ma- tsuda 13)&@A "#''" #(( @A S. au- reus ''"#)B"1& "1& 2) 1 4 VII IVH 6hiWX 3 MPIPC [ *C Tenover 14) PFGE D7E 4 8 mg/ml borderline-resistant S. aureus +FG,HI- '#D "1&K mecA PBP2῎ K3& 7E.JK3LMNO67 2 3 D7E MSSA % PQ"/(R01S '+ FG,1 4 T= !"# S. aureus 3) PFGE 7 20 S. aureus Y4567 /(2R0KUV 4 WX Y+>$?"8ZN aureus '' S. au- @A S. O67UV [34 \56A 73] reus K1S"# S. aureus % @A WX S. aureus K ^_8[G9`^aV b$:;1& cc @A S. aureus K @A S. aureus Kj ^_g< NO67 S. aureus =h> de^_ f]+-#.>$ ?@ ? S. aureus gg<<=h>?@ =h$R0K[ Gs$Z K @A MRSA AB S. aureus ijC 9 MRSA ?B!HI"1 kK cKlmDnE- S. aureus MRSA KF9op 60q70r *C1) tuv MRSA Gs$ 8GwBxHI3yB"1 Q X# Js^_"#zKL{M S. aureus Kx8[2), 12) @A S. aureus Kx8[ #S S. aureus K@ANOJ8 2) Q3 "3UV MRSA $|P} mD1~_QR$"1 $|PS89?B_1$ _TcmD((& UV(N WX,YZ[KHI"1UV Q ῎ ῍ 1) fg$:!*Ck 2000. l 11 cfmn~ I-10῍II-41. ¡mDo*¢ £¤p¥756 2) q¦r? 2002. KL!st § u¨ GwBῌKL{M v^yB>wx ῌ KLefg 29: 253῍260. 3) f©? 1991. MRSA yª« p. 78῍130, MRSA $ 8 Z ¬ z { | }¡ m ®6¯° ~± 4) c² 2002. QR$ (1) ³´¢7yµ ¶E·¸B p. 100῍116, $[9 ?¹J ³º» ~± 5) ¼ ? 2000. II !-L 29 <=>?tuvwx0ty! #tz!{ht|D}h 158 6) 7) 8) 9) 10) 1. Methicillin-resistant Staphylococcus aureus (MRSA). p. 40῍47, Yearbook 2000. 111 !"# 2002. $%&'( p. 12῍29, ) *+,-./01234 56789 : ; National Committee for Clinical Laboratory Standard. 2000. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically Aproveded standardῌ5th ed. M7-A5. NCCLS, Wayne, USA. Tokue, Y., S. Shoji, A. Watanabe, et al. 1992. Comparison of a polymerase chain reaction assay and a conventional microbiologic method for detection of methicillin-resistant Staphylococcus aureus. Antimicrob. Agents Chemother. 36: 6῍9. <=>? @!AB CDEF G 1995. PCR HIJ population analysis KILMNOPheterogeneous MRSA. 43: 487῍492. Nakatomi, Y., J. Sugiyama. 1998. A rapid latex 11) 12) 13) 14) 15) agglutination assay for the detection of penicillin-binding 2῎. Microbiol. Immunol. 42: 739῍ 743. Louie, L., O. S. Matsumura, E. Choi, et al. 2000. Evaluation of three rapid methods for detection of methicillin resistance in Staphylococcus aureus. J. Clin. Microb. 38: 2170῍2173. QRST 2002. UVWXYZ [\] 29: 275῍277. Matsuda, J., Y. Hirakata, F. Iori, et al. 1998. Genetic relationship between blood and nonblood isolates from bacteremic patients determined by pulsed-field gel electrophoresis. J. Clin. Microbiol. 36: 3081῍3084. Tenover, C. F., D. R. Arbert, V. R. Goering, et al. 1995. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J. Clin. Microbiol. 33: 233῍2239. :^_ ` 2000. a)*bcde p. 245῍ 253, )*f ghi j kilmno pqr5 678sr Studies of Clinical Microbiology and Molecular Biology of Staphylococcus aureus Isolated from Blood Culture and Non-blood Specimens of the Same Patients Shoji Moriki,1) Akiko Nobata,1) Hiroshi Shibata,1) Junichi Masuda2) and Shunichi Kumakura3) 1) 2) Shimane University Hospital Central Clinical Laboratory Shimane University School of Medicine Laboratory Medicine and Central Clinical Laboratory 3) Shimane University Hospital Division of Blood Transfusion From July to December 2002, we found 7 bacteremic patients who were positive for Staphylococcus aureus in both blood and non-blood isolates. In total, 20 isolates were studied to examine the phenotypic and genetic relationship between blood and non-blood isolates in each patient. Pattern of antibiograms and the coagulase types of S. aureus from blood specimens were consistent with those from the non-blood specimens in each patient. Two patients had MSSA, out of which borderline-resistant S. aureus was identified in 1 case, and 5 patients had MRSA. Genetic analysis by pulsed-field gel electrophoresis (PFGE) showed the same patterns between blood and non-blood specimens in individual cases. However, in 1 case, di#erent PFGE patterns were observed between 2 blood isolates that were cultured under aerobic and anaerobic conditions. Our present results indicate that S. aureus strains obtained from blood and non-blood isolates were phenotypically and genetically identical in each patient, suggesting the possibility that the same clone of S. aureus could be systemically expanded in cases of bacteremia caused by this pathogen. Key words: blood culture, borderline-resistant S. aureus, PCR (polymerase chain reaction), PFGE (pulsedfield gel electrophoresis), contact transmission 30
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