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MRSA $ 8 Z Ÿ¬ z { | }¡ m ­®6¯”° ~±š
4) €c²š 2002. QR$ (1) ³´¢ˆ7yµ
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1. Methicillin-resistant Staphylococcus aureus (MRSA). p. 40῍47, Yearbook 2000. 111 !"# 2002. $%&'( p. 12῍29, )
*+,-./01234 56789 :
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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