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1 Key words : Burkholderia cepacia, -PCR method, intrahospital transmission
2
3 690 岡崎 充宏 他 Fig. 1 The epidemiology of B. cepacia in the Kyorin Month ず つ 分 離 さ れ た. 3. のDNAお よ び 各 キ ッ トに -PCR法 ン は,2種 に よ る 供 試 菌 株 のDNAパ ター 類 の プ ラ イ マ ー を 同 時 に使 用 した 結 果 主 要 な5つ 2 の 型 を 示 し た(Fig.2).さ の パ タ ー ン の 中 のR3は,約300bpの 有 無 に よ り2つ R3b)に 区 別 さ れ た(Table, surgery ら に これ ら 分 子 量 の の パ タ ー ン(R3aお 示 し て い な い が,1種 (RPKHM1)を R1お 1).な お,結 果 には ward) R2 base size b marker. (TCC) 4, isolates. R1 R5 3, isolate j (thoracic (TCC) 5, isolate m (respi- I digest on the a M,ƒÓ ~174/Hinf Numbers isolate i Lane 2, isolate R3 cepacia ward) B. ward) R4 DNA of left for are pairs. タ ー ン は, の 型 し か得 られ な か っ た. こ れ ら の 主 要 な5つ 析 は,1996年1月 のDNAパ か ら2月 タ ー ンか らの 解 に か け てTCCを に 分 離 さ れ た20菌 株 のDNAパ ザ ー 由 来 の2菌 よび. 類 の み の プ ラ イ マ ー 使 用 し た 時 のDNAパ よ びR2の2つ s ratory DNAの 1, isolate (respiratory よ る表 現 型 を 指標 と した 型 別 の解 析 Hospital of isolation Fig. -PCR法 University 株 と一 致 し た.ま 外 の 菌 株 のDNAパ 解 析 の 再 現 性 は,供 果,各 菌 株 のDNAパ た,そ タ ー ン は,そ タ ー ン と 異 っ た(Table 中心 ター ン が ネ ブ ラ イ 1).本 の期 間以 れ ら20菌 株 の パ 方 法 に よ るDNA 試 菌 株 を繰 返 し3回 タ ー ン が3回 行 った 結 と も 一 致 し,安 定 し て い た. 一方,各 キ ッ トに よ る 表 現 型 別 の 解 析 は,ID-18 とAPIで は そ れ ぞ れ1お よ び2パ タ ー ン を示 し, ネ ブ ラ イ ザ ー 由 来 株 のパ ター ン は 臨 床 材 料 由 来 株 と 一 致 し た.ま た,NC4Jで が,患 者Cか の 中 で4パ は11パ タ ー ン を 示 し た ら 数 回 に わ た っ て 分 離 さ れ た6菌 タ ー ン が 認 め ら れ た(Table 株 1). 感 染症 学 雑 誌 第72巻 第7号 in
4 Table 1 Origins and phenotypic and genotypic characteristics of the B. cepacia isolates
5 Pseudornonas aeruginosa and Burkholderia ce- pacia. Microbiol Rev 1996 ; 60 : ) Jack DS, Nehama H, Gabi R, David M : Nosocomial Pseudomonas cepacia infection associated with chlorhexidine contamination. Am J Med 1982 ; 73 : ) Spencer RC : The emergence of epidemic, multiple-antibiotic-resistant Stenotrophomonas (Xanthomonas) maltophilia and Burkholderia (Pseudomonas) cepacia. J Hosp Infect 1995 ; 30 : ) Hoiby N : Isolation and treatment of cystic fibrosis s with lung infections caused by Pseudomonas (Burkholderia) cepacia and multiresistant Pseudomonas aeruginosa. J Med 1995 ; 46 : ) David AP, Daniel VS, Ofclia CT, Loretta AC, Nancye CC, William RJ : Possible nosocomial transmission of Pseudomonas cepacia in s with cystic fibrosis. Arch Pediatr Adolesc Med 1994 ; 148: ) Richard JH, Eric DH, Paul RG et al.: An outbreak of Burkholderia (formerly Pseudomonas) cepacia respiratory tract coloni- and infection associated with nebulized zation albuterol therapy. Ann Intern Med 1995 ; 122 : ) Pegues CF, Pegues DA, Ford DS et al.: Burkholderia cepacia respiratory tract acquisition : Epidemiology and molecular characterization of a large nosocomial outbreak. Epidemiol Infect 1996 ; 116: ) Rabkin CS, Jarvis WR, Anderson RL et al.: Pseudomonas cepacia typing systems : Collabor- ative study to assess their potential in epidemiologic investigations. Rev Infect Dis 1989 ; 11 : ) Daniel JA, Janet SK, Michael LV, Dale NG, Edward NJ : DNA fingerprinting by pulsed field gel electrophoresis and ribotyping to distinguish 1) Beryl JR, David EH : Pneumonia and septicemia Pseudomonas cepacia isolates a due to Pseudornonas cepacia in a nosocomial outbreak. J Clin Microbiol 1991 ; with cystic fibrosis. Johns Hopkins Med 29 : ) Eshwar M, Maureen EC, Deborah AH, David J 1980 ; 147: ) Mary C, Vernon F : Determinants of mortality cystic fibrosis in Canada, Am J Epidemiol 1996 ; 143: ) Govan JRW, Deretic V : Microbial pathogenesis in cystic fibrosis : Mucoid PS : Epidemiology of Burkholderia cepacia infection in s with cystic fibrosis : Analysis by randomly amplified polymorphic DNA fingerprinting. J Clin Microbiol 1996 ; 34 :
6 14) Stauffer GH, Plamann MD, Stauffer LT : Construction and expression of hybrid plasmids containing the Escherichia coli gyl A gene. Gene 1981 ; 14 : genotypic heterogeneity of multidrug-resistant Pseudomonas aeruginosa serotype O: 12 out break isolates a pediatric hospital. J Clin Microbiol 1996 ; 34 : ) Tambic A, Power EGM, Talsania H et al. : Analysis of an outbreak of non-phage-typeable 17) Kerr JR, Moore JE, Curran MD et al. : Investigation of a nosocomial outbreak of using a randomly amplified polymorphic DNA methicillin-resistant Staphylococcus aureus by Pseudomonas aeruginosa pneumonia in inten- assay. J Clin Microbiol 1997 ; 35 : sive care unit by random amplification of polymorphic DNA assay. J Hosp Infect 1995 ; 30 : ) Edouard B, Stephane B, Pierre R et al. : Molecular epidemiology provides evidence of Analysis of Transmission of Burkholderia cepacia Isolates in an Intrahospital by Randomly Amplified Polymorphic DNA-PCR Method Mitsuhiro OKAZAKI1), Koji MORITA2), Naoko KOCHI1), Koji ARAKI1), Mie YOSHIZAWA3), Hiroshi WADA2), Mikiyoshi SHIBATA2), Noboru WATANABE2), Teruo EGAMI1), Nobushige FURUYA1), Masato KANAMORI2), Shuji SHIMAZAKI3) & Hidemasa UCHIMURA1) of Clinical Laboratories, Kyorin University Hospital 2)Department of Microbiology, Kyorin University School of Health Sciences 3)Trauma and Critical Care Center, Kyorin University Hospital Strains of Burkholderia cepacia isolated in our hospital November 1995 to September 1996 were classified with randomly amplified polymorphic DNA-PCR (-PCR) and conventional biochemical tests (ID test E NF-18, API2ONE, and Neg Combo 4J kit), and intrahospital isolates of B. cepacia were analysed. During the period 28 strains ins and 2 medical apparatus were isolated. Twenty four of 28 (85.7%) were sputum. In 1996 January to February, 20 strains were detected 8 ins, and two strains were the nebulizers at the Trauma and Critical Care Center (TCC). With typing of B. cepacia by conventional methods no epidemiological relations among isolates were found. However, DNA patterns of original isolates the nebulizers at TCC by -PCR were identical with those of isolates in sputa s in other wards who had stayed at TCC, indicating that TCC was an initial source of transmission and the strain was transmitted with the s to the wards. These results suggest that -PCR method might be an useful tool to analyse an epidemiological survey for intrahospital transmission of isolate.
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