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이유철,설성용,조동택,전도기,Lee, Yoo-Chul,Seol, Sung-Yong,Cho, Dong-Taek,Chun, Do-Ki 대한미생물학회 1987 大韓微生物學會誌 Vol.22 No.1
Multiply resistant Shigella strains isolated in Taegu area were subjected for the characterization of R plasmids. All strains isolated in 1984 and 1985 were susceptible to gentamicin, amikacin, and cephalothin, and most strains were susceptible to kanamycin (Km) and rifampin by agar dilution antimicrobial susceptibility test. The resistance frequency of S. flexneri against ampicillin (Ap) was higher than that of S. sonnei. The strains resistant to sulfisomidine (Su) and trimethoprim (Tp) were found at higher frequency in S. sonnei than in S. flexneri. The most prevalent resistance pattern of S. flexneri was chloramphenicol (Cm) tetracycline (Tc) streptomycin (Sm) Ap, followed by the pattern of CmTcSmSuApTp, CmTcSmSuApTp nalidixic acid, and CmTcSmSuAp in the decreasing order. The antibiogram of CmTcSmSuTp was found to be the most frequent pattern in S. sonnei. The ratio of conjugal transfer of S. flexneri was 47% and 75% of S. sonnei. The average number of plasmid harboring in Shigella was 4 and the size of plasmid ranged 1.3 to 134 megadalton (Mdal). Most S. flexneri carried plasmids of 2 to 3 Mdal and S. sonnei carried those of 3 to 4 Mdal size. The sizes of conjugative plasmids ranged 40-90 Mdal. The incompatibility group (Inc) F II plasmids (54-59 Mdal) were most frequent and rare Inc B plasmids (60 Mdal) of isolates in 1979 and 1980 and Inc FI (87 Mdal) of 1983 isolates were able to be classified by the colony test with standard reference plasmids. The R plasmids of known Inc group were tested for the restriction endonuclease analysis. The pattern of plasmids digested by EcoRl were apparently different by the Inc group but there was no significant difference between species or by the resistance patterns. Nonconjugative plasmids and their phenotypes were identified by transformation test. The transformants were resistant to less than two drugs. Colicin producing transformants carried the Col plasmid of 3.7 or 3.9 Mdal size. $Ap^r$ plasmids derived from S. sonnei were found to be mobilized by transfer factor RT641 to E. coli #CS100. $Ap^r$ plasm ids of same size shared by S. flexneri, S. sonnei, and E. coli were digested with Pstl. All of them showed two restriction fragments of 2.8 kilobase(kb) and 0.7kb. Other plasmids ($Sm^r\;Su^r$) derived from S. flexneri, S. boydii, and S. sonnei were digested with Pstl and they showed same restriction fragment patterns of 3.1kb and 2.9kb. The plasmid profiles of three strains of S. sonnei producing colicin and showing same resistance pattern of CmTcSmSuApTpKm appeared to be similar. Restriction patterns by EcoRl and the behavior of plasmids in conjugation or transformation process were also similar between those plasmids. The restriction patterns were significantly different between the plasmids of Inc FI group and those of unclassified Inc group.
$bla_{SHV-2a}$와 $bla_{SHV-12}$ 항균제 내성 유전자의 분자적 진화 및 확산에 IS26 Mobile Element의 개입
김정민,신행섭,조동택,Kim, Jung-Min,Shin, Haeng-Seop,Cho, Dong-Taek 대한미생물학회 2000 大韓微生物學會誌 Vol.35 No.3
A clinical isolate of Klebsiella pneumoniae K7746 produced the extended-spectrum ${\beta}$-lactamase (ESBL) SHV-12. A 6.6 kb BamHI fragment containing the $bla_{SHV-12}$ gene of K7746 strain was cloned into pCRScriptCAM vector resulting in the recombinant plasmid p7746-Cl. The restriction map of 3.6 kb inserted DNA and sequences immediately surrounding $bla_{SHV-12}$ of p7746-C1 were homologous to plasmid pMPA2a carrying $bla_{SHV-2a}$. In addition, both $bla_{SHV-12}$ and $bla_{SHV-2a}$ were expressed from a common hybrid promoter made of the -35 region derived from the left inverted repeat of IS26 and the -10 region from the $bla_{SHV}$ promoter itself. The results indicate that $bla_{SHV-12}$ and $bla_{SHV-2a}$ may have evolved from a common ancestor in the sequential order of $bla_{SHV-2a}$ first, followed by $bla_{SHV-12}$. Furthermore, by the PCR mapping method using primers corresponding to the IS26 and $bla_{SHV}$, the association between IS26 and $bla_{SHV}$ was studied in 12 clinical isolates carrying $bla_{SHV-2a}$, 27 clinical isolates carrying $bla_{SHV-12}$, and 5 reference strains carrying $bla_{SHV-1}$ to $bla_{SHV-5}$. All 39 strains carrying $bla_{SHV-2a}$ or $bla_{SHV-12}$ were positive by the PCR, providing confirmative evidence that IS26 has been involved in the evolution and dissemination of $bla_{SHV-2a}$ and $bla_{SHV-12}$. But 5 reference strains carrying $bla_{SHV-1}$ to $bla_{SHV-5}$ were negative by the PCR. Therefore, we concluded that the molecular evolutionary pathway of $bla_{SHV-2a}$ and $bla_{SHV-12}$ may be different from that of other $bla_{SHV-ESBL}$, e.g., $bla_{SHV-2}$, $bla_{SHV-3}$, $bla_{SHV-4}$, and $bla_{SHV-5}$.
Prevalence and Drug Resistance of Shigella in Taegu Area of Korea
전도기,박종욱,서성일,조동택,설성용,이유철,Chun, Do-Ki,Park, Jong-Wook,Suh, Seong-Il,Cho, Dong-Taek,Seol, Sung-Yong,Lee, Yoo-Chul The Korea Society for Microbiology 1986 大韓微生物學會誌 Vol.21 No.4
1973년부터 1985년 사이에 대구지방에서 분리된 Shigella는 약 1,200주였는데, 대부분이 Shigella flexneri였고, S. sonnei는 약 20%였으며, S. dysenteriae와 S. boydii는 극히 적었다. 분리균의 95%이상이 chloramphenicol(Cm), tetracycline(Tc), streptomycin(Sm), sulfisomidine(Su), ampicillin(Ap), trimethoprim(Tf) 등의 전부 또는 일부 약제에 내성이었으며, kanamycin, nalidixic acid(Na)와 rifampin(Rf)에 내성인 균주는 소수 있었으나, cephaloridine, gentamicin, amikacin 등에 내성인 균주는 없었다. 1973년에 분리된 균은 약 절반이 약제내성이었으나, 1977년 이후 분리된 균은 95% 이상이 약제내성이었다. 1977년 까지는 Cm, Tc, Sm 및 Su의 4제 내성균이 가장 많았으나 1978년 부터는 Cm, Tc, Sm, Su, Ap 및 Tp의 6제 내성균이 가장 많았다. 약 75%의 균이 그 약제내성을 접합에 의하여 Escherichia coli에 전달하였으므로 이 약제내성은 R plasmid에 의한 것으로 생각되었다. 대다수의 균에 있어서 약제내성 전부를 E. coli에 전달하였으나, Na와 Rf의 내성은 전달되지 않았다. 일부 균주의 약제내성은 E. coli에 전달될 때 분리되는 일이 있었으나, 그 수는 극히 적었다. Shigella의 R Plasmid는 대부분이 비적합성군(incompatibility group) F II에 분류되었으나, 극소수는 B군에 속하였고 군별이 안되는 것도 있었다. Shigella strains isloated in the Teagu area during the period from 1973 to 1985 were studied for species distribution, drug resistance, and R plasmids. Approximately 1,200 strains were isolated during this period, and most of them were classified into Shigella flexneri, S. sonnei occupied less than 20%, and S. dysenteriae and S. boydii were very rarely isolated. More than 95% of them were resistant to one or more of these drugs; chloramphenicol (Cm), tetracycline (Tc), streptomycin (Sm), sulfisomidine (Su), ampicillin (Ap), and trimethoprim (Tp). Strains resistant to kanamycin, nalidixic acid (Na), and rifampin (Rf) were rare, and no strain was resistant to cephaloridine, gentamicin, and amikacin. Approximately half of the isolates were resistant to drugs in 1973, but the rate of resistant strains increased to more than 95% from 1977. Strains resistant to the four drugs (Cm, Tc, Sm, and Su) occupied the majority of resistant strains until 1977, but the most prevalent multiplicity of drug resistance increased to six drugs (Cm, Tc, Sm, Su, Ap, and Tp) from 1978 with the marked increase of Ap- and Tp-resistant strains. Approximately 75% of them transferred resistance to Escherichia coli by conjugation, and the resistance was considered to be mediated by R plasmids. Almost all of them transferred the complete patterns of resistance to drugs except Na and Rf. However, among some strains of recent isolates, small numbers of segregants of transferred resistance were observed. The R plasmids in Shigella were mostly classified into Inc FII, and only small numbers into Inc B. Segregants were in most cases unclassified.