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국내 대학병원에서 분리된 Metallo-β-Lactamase (MBL) 생성 Acinetobacter spp. 분리주의 높은 출현율과 유전형 특징
염종화 ( Jong Hwa Yum ) 대한임상검사과학회 2019 대한임상검사과학회지(KJCLS) Vol.51 No.4
주요 획득성 metallo-β-lactamase (MBL) 유전자에 의해 매개되는 carbapenem 내성, 특히 Acinetobacter spp. 균종의 임상 분리주에 대한 보고가 증가하고 있다. 본 연구에서 임상에서 비 중복으로 분리된 carbapenem 비감수성 Acinetobacter spp. 191주 중 125 (65.4%)주가 imipenem 혹은 meropenem-Hodge 변법시험에 양성이었고, 49 (25.7%)주가 imipenem-EDTA+SMA double disk synergy (DDS) 시험에 양성이었다. bla<sub>VIM-2</sub> allele와 bla<sub>IMP-2</sub> allele 검출을 위한 중합효소연쇄반응과 염기서열분석을 시행한 결과, A. baumannii 와 A. calcoaceticus에서 각각 29주와 1주가 bla<sub>VIM-2</sub>를 갖고 있었고, A. baumannii 16주와 A. calcoaceticus 2주가 bla<sub>IMP-1</sub>을 갖고 있었다. A. genomospecies 3는 bla<sub>VIM-2</sub>와 bla<sub>AIM-1</sub>을 동시에 갖고 있었다. 이들 MBL 유전자는 모두 class 1 integron에 있었다. bla<sub>VIM-2</sub> 혹은 bla<sub>IMP-6</sub>를 갖는 class 1integron의 크기는 A. baumannii 분리주에서는 2.8 kb에서 3.2 kb이었고, A. genomospecies 3 분리주에서는 3.2 kb에서 3.5 kb이었다. bla<sub>VIM-2</sub>는 대부분 class 1 integron에 첫번째 혹은 두번째에 위치하였고, aacA4를 흔히 가지고 있었다. 다양한 내성 유전자를 가질 수 있는 MBL 생성 Acinetobacter spp. 뿐 아니라 다양한 내성 유전자를 가질 수 있는 integron의 전파로 imipenem이나 meropenem과 같은 carbapenem 내성을 포함하여 다제 내성 그람음성 세균의 증가가 예상된다. 또한, 위중한 Acinetobacter spp. 감염증 치료를 위한 새로운 항균제 개발이 필요하다. Carbapenem resistance, mediated by the major acquired metallo-β-lactamase (MBL) genes, has been increasingly reported, particularly for clinical isolates of Acinetobacter spp. Of the 191 nonduplicate clinical isolates of the carbapenem-nonsusceptible Acinetobacter spp. evaluated, 125 isolates (65.4%) were positive for the modified imipenem or meropenem-Hodge test, and 49 isolates (25.7%) were positive for the imipenem-EDTA+SMA double disk synergy test (DDS). PCR and sequencing of the bla<sub>VIM-2</sub>-allele and blaIMP-1-allele showed that 29 A. baumannii isolates and 1 A. calcoaceticus isolate had bla<sub>VIM-2</sub>, whereas 16 A. baumannii isolates and 2 A. calcoaceticus isolates had bla<sub>IMP-6</sub>; 1 isolate of the A. genomospecies 3 had bla<sub>VIM-2</sub> and bla<sub>AIM-1</sub>. All the above MBL genes belong to class 1 integron. The size of class 1 integron encompassing bla<sub>VIM-2</sub> or bla<sub>IMP-6</sub> ranges from 2.8 kb to 3.2 kb in clinical isolates of A. baumannii, and 3.2 kb to 3.5 kb in clinical isolates of A. genomospecies 3. bla<sub>VIM-2</sub> was most often located first or second in the class 1 integron, and these integrons often included aacA4. Due to dispersion of the MBL-producing Acinetobacter spp. as well as integron, which may encompass various resistance genes, there is an expectation for the increase of multidrug resistant Gram-negative bacteria, including resistance of carbapenems such as imipenem or meropenem. Hence, the development of new antimicrobial agents for treating severe Acinetobacter spp. infections is needed.
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지역사회 협의체 중심의 지역아동센터 구강건강증진 프로그램의 효과
염종화 ( Jong Hwa Yum ),김혜진 ( Hye Jin Kim ),권명화 ( Myoung Hwa Kwon ),신선정 ( Sun Jung Shin ) 한국치위생과학회 2014 치위생과학회지 Vol.14 No.2
This study measures the effect of oral health promotion program based on community networking for elementary school students in community child center. The community networking were constructed of community health center, headquarters for community child center and school of dental hygiene in community. First, we were educated the student and teacher of community child center, separately. Community health center planned and evaluated the program, and school of dental hygiene ran the maintenance program once a month for 3 months and evaluated the program. The teacher of community health center were supported and monitored the children. The comparison was done in independent t-test of awareness, knowledge and behavior of children of community child center in both lower grades and upper grades and paired t-test of patient hygiene performance (PHP) index was conducted before and after oral health promotion program. As a result, PHP index and oral health knowledge increased significantly after oral health program in lower grades and upper grades (p<0.001). The positive attitude for oral health about .Whatever I do, my tooth-will be decayed. increased more in upper grades better than lower grades after oral health promotion program (p<0.05). We suggest that oral health program based on community networking should be constructed for oral health promotion of elementary school students in community child center.
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16S rRNA에 의한 한국 내 Chyseobacterium indologenes과 염기 서열 변화
Man Kyu Huh(허만규),So Hye Park(박소혜),Jong Hwa Yum(염종화) 한국생명과학회 2011 생명과학회지 Vol.21 No.6
병원균에 대한 정확한 동정은 임상 연구실에서 필수적인 요소의 하나이다. Chyseobacterium indologenes에 대한 동정을 포함한 분자생물학적 분석과 리보솜의 16S rRNA 유전자로 한국에서 추출한 17검체와 GenBank에서 Chyseobacterium속 검색을 통해 이들과 계통관계를 평가하였다. C. indologenes의 배당 서열은 1,176 nucleotides였다. C. indologenes 내의 서열 변이는 주로 염기 치환이었다. 한국의 C. indologenes 검체는 다른 나라의 동 종과 크게 다르지 않았다. 그런데 한국의 C. indologenes의 치환율은 GenBank에 있는 동종보다 높았다. C. indologenes는 C. isbiliense, C. hominis, C. hispanicum, C. molle, C. hungaricum, and C. pallidum과 자매종을 형성하였다. Accurate identification for pathogenic bacterium is an essential element in the clinical microbiology laboratory. We studied molecular analysis involving the identification of Chyseobacterium indologenes and evaluated the seventeen isolates in Korea with the 16S rRNA gene of the ribosome to estimate phylogenetic relationships within the genus Chyseobacterium in GenBank. The aligned data sets for C. indologenes were 1,176 nucleotides. Sequence variation within the C. indologenes was mostly due to nucleotide substitutions. Korean C. indologenes isolates were not strikingly different from the same species found in the other countries. However, the rates of base substitution in Korean C. indologenes isolates were higher than those of other C. indologenes isolates in GenBank. C. indologenes was placed as a sister species to C. isbiliense, C. hominis, C. hispanicum, C. molle, C. hungaricum, and C. pallidum.
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연구논문 : 오배자(Rhus javanica) 추출물의 어병세균 Vibrio ichthyoenteri 와 Streptococcus iniae 에 대한 항균활성
김경훈 ( Kyoung Hoon Kim ),김아라 ( Ah Ra Kim ),조은지 ( Eun Ji Cho ),주성제 ( Seong Je Joo ),박종훈 ( Jong Hoom Park ),문지영 ( Ji Young Moon ),염종화 ( Jong Hwa Yum ),김태훈 ( Tae Hoon Kim ),권현주 ( Hyun Ju Kwon ),이은우 ( Hyun 한국수산과학회(구 한국수산학회) 2014 한국수산과학회지 Vol.47 No.1
The antibacterial activities of methanol extracts of 19 commercial herbal medicines were measured against the fish pathogens Vibrio ichthyoenteri and Streptococcus iniae, which cause several fish diseases. Rhus javanica showed the strongest antibacterial activity against V. ichthyoenteri and S. iniae. The methanol extract of R. javanica was extracted further using several organic solvents with different polarities. The extract from the ethyl acetate fraction showed strong activity against both fish pathogens. The minimum inhibitory concentration (MIC) of the R. javanica extract was 32μg/mL for V. ichthyoenteri and 128μg/mL for S. iniae. Further purification and isolation of the active com- pound (s) responsible for these activities and further study of the synergistic effect using combinations of antibiotics against pathogenic bacteria are needed.
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Bacillus amyloliquefaciens H41이 생산하는 Vibrio anguillarum 생육 저해인자의 정제
신현철(Hyun-Chul Shin),정경태(Kyung-Tae Chung),김광현(Kwang-Hyun Kim),김병우(Byung-Woo Kim),권현주(Hyun-Ju Kwon),이은우(Eun-Woo Lee),염종화(Jong-Hwa Yum),류은주(Eun-Ju Rhu),정유정(Yu-Jeong Jeong),김영희(Young-Hee Kim) 한국생명과학회 2008 생명과학회지 Vol.18 No.6
어류 질병 치료를 위한 probiont의 개발에 목적을 두고 어병균 Vibrio anguillarum NCMB1의 생육에 저해물질을 생산하는 Bacillus amyloliquefaciens H41균주를 분리하고 이 물질의 특성을 규명하기 위하여 정제를 시행하였다. 분리 균을 배양한 배양 상등액을 70% 염석, 투석하여 조 효소액으로 제작하고 조 효소액을 DEAE-sephadex, A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography을 통하여 정제하고 SDS-PAGE 를 통하여 단일밴드를 확인하고 최종 회수율 2.9%을 얻을 수 있었으며 40.8배의 정제된 V. anguillarum NCMB1 생육저해물질을 얻을 수 있었다. 정제된 저해물질은 저해정도에 따라 단위를 설정하여 활성을 측정하였으며, 분자량은 48 kDa 로 확인되었으며 정제물질의 활성을 위한 최적 반응 pH와 온도는 pH 7.5와 30℃로 확인되었다. 금속이온의 효과에 있어서는 CoCl₂, HgCl₂, ZnSO₄, AgNO₃에서는 완전히 저해되는 양상을 나타내었고 MgSO₄, MnSO₄에서 미미한 효소활성의 증가를 나타내었다. 그리고 염에 관한 안정성은 일반 해수의 농도인 3%의 농도에서도 활성을 나타내는 것으로 확인되었다. 정제된 저해물질을 현재 상업적으로 사용하고 있는 화학처리제나 항생제와 함께 효율성을 비교 해 보았을 때 저해물질은 약 78%의 저해 활성을 나타내는 것을 알 수 있었고 항생제보다는 효율성이 낮았으나 독성검사를 위해 정제물질을 살아있는 어류에 투여하였으나 어떤 해수어도 폐사하지 않는 것으로 보아 어류 자체엔 독성을 나타내지 않는 물질로 나타났다. 따라서 B. amyloliquefaciens H41 균주가 생산하는 정제 물질이 V. anguillarum 생육에 저해물질로 작용하는 것으로 밝혀졌으며 친환경적인 특성을 가진 물질로 밝혀졌다. To study the possible use of probiotics in fish farming, we evaluated antagonism of antibacterial strain Bacillus amyloliquefaciens H41 against the fish pathogenic bacterium Vibrio anguillarum NCMB1. The purification of growth inhibition factor produced by B. amyloliquefaciens H41 was achieved by obtaining supernatant of this bacterium. The growth inhibition factor was purified to homogeneity by 70% ammonium sulfate precipitation, DEAE-sephadex A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography, and sephadex G-50 gel filtration column chromatography with 40.8 fold of purification and 2.9% yield. The molecular weight of the purified growth inhibition factor was 48 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH and temperature for the growth inhibition factor were pH 7.5 and 30℃, respectively. The activity of growth inhibition factor was enhanced slightly by some metal ions, such as Mg?², Mn?², but was inhibited by the addition of Co?², Hg?², Zn?² and Ag?². NaCl stability of the growth inhibition factor was observed with 50% residual activity at 3% NaCl concentration. Toxicity test showed that the purified B. amyloliquefaciens H41 growth inhibition factor did not affect the live of Japanese flounder (Paralichthys olivaceus) and the effectiveness was 78% of residual lethality compared to commercial antibacterial agents.
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Amoxicillin-Sulbactam (Sultamox)의 주요 임상 분리세균에 대한 시험관내 항균력
염종화,용동은,김명숙,이경원,정윤섭 대한화학요법학회 2002 대한화학요법학회지 Vol.20 No.3
목적 : β-lactam 항균제는 감염증 치료에 가장 흔히 사용되는 항균제 중의 한가지이다. 그러나 최근 임상검체에서 분리되는 세균중에는 β-lactamase를 생성하는 세균이 증가하는 추세로 세균 감염증 치료시 penicillin 및 amoxicillin 제제의 유효성이 현저히 감소 하였고, 이에 새로운 약제의 개발이 필요하게 되었다. 이 연구에서는 amoxicillin 와 β-lactamase 저해제를 병합한 AXS의 시험관내 항균력을 시험하고자 하였다. 방법 : 2000 2001년 세브란스병원 환자의 임상검체에서 분리된 각종 호기성 세균에 대한 항균체 감수성을 NCCLS 한천희석법으로 시험하였다. 결과 : Oxacillin 감수성인 S. aureus와 coagulase 음성 Staphylococcus, S. pneumoniae 및 S. pyogenes는 시험균주 모두가 APS, AXC 및 AXS에 감수성이었다. M. (B.) catarrhalis와 H. influenzae에 대한 MIC_90는 amoxicillin이 〉128 ㎍/mL이었으나, APS, AXC 및 AXS는 각각 8-32 ㎍/mL과 4-16 ㎍/mL이었다. 결론 : AXS은 oxacillin 감수성 Staphlococcus, M. (B.) catarrhalis, H. influenzae, K. oxytoca 및 P. mirabilis에 대한 시험관내 항균력이 우수하여 이들 세균에 의한 감염증 치료에 유용하게 사용될 수 있을 것으로 판단된다. Background : β-lactam antibiotics are one of the most frequently used antimicrobial agents. However, with the increase of β-lactamase-producing bacteria, penicillins and amoxicillin have become less useful. Amoxicillin-sulbactam (AXS) was developed to restore the activity. The aim of this study was to determine the in vitro activities of AXS against recent clinical isolates. Methods : Various aerobic bacteria tested were isolated from clinical specimens in Severance Hospital during 2000 to 2001. Antimicrobial susceptibility was determined by the NCCLS agar dilution method. Results : Oxacillin-susceptible Staphylococcus aureus and coagulase-negative Staphylmoccus, Streptococcus pneumoniae and S. pyogenes were susceptible .to ampicillin-sulbactam (APS), amoxkillin-clavulanic acid (AXC) and -AXS. For Moraxella ('Branhameh'a) catarrhah and Haemophdm infuenzae, MIC_90s of Aps, AXC and ,AXS were 0.5' 3.lg/mL and 8 ㎍/mL, respectively, which were &fold lower than those of amoxicillin For Klebsiella oxytoca and Proteus mirabilis, MIC_90 of amoxkillin was >128 ㎍/mL, but MIC_90S of AFS, AXC and AXS were 8-32 ㎍/ml and 4-16 ㎍/ml, respectively. Conclusion : AXS showed good antimicrobial activities against oxacillin-susceptible staphylococcus, M (B..) catarrhalis, H influenzae: K oxytoa and Mirabilis. Therefore, It would be useful for the treatment of infections due to these organisms.
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Mecillinam의 임상분리 장내세균 대한 시험관내 항균력
김창기,염종화,이상국,이양순,최준용,김준명,이경원,정윤섭 대한감염학회 2009 감염과 화학요법 Vol.41 No.3
Background : Mecillinam, an amidinopenicillin antibiotic, has been used to treat urinary tract infections and bacterial enteritis in many countries, In this study, we evaluated in vitro activity of mecillinam against Enterobacteriaceae isolates from urine, and Salmonella and Shigella isolates from patients with bacterial gastroenteritis. Materials and Methods : A total of 308 clinical strains were collected and were comprised of Escherichia coli (n=109), Klebsiella pneumonias (n=52), Enterobacter spp. (n=30), Serratia marcescens (n=30) and Proteus spp. (n=29) isolated from a university hospital in Korea in 2007, and of Salmonella spp. (n=28) and Shigella spp. (n=30) isolated from Korean diarrheal patients from 2001 to 2006. Antimicrobial susceptibility was tested by Clinical laboratory Standard Institute (CLSI) agar dilution method. CLSI breakpoint of mecillinam for E. coli urinary tract isolates was applied to all other isolates. Results : In E. coli, rate of susceptibility to ampicillin was 30%, but 99-100% to amikacin and cefotaxime. Most (96%) of E. coli isolates, including extended-spectrum B-lactamase (ESBL) Producers, were susceptible to mecillinam. All ESBL producers, except for one isolate, were inhibited by ≤4 ug/mL of mecillinam. MIC_90 of mecillinam for K. pneumonias and Enterobacter spp. was 8 ug/mL and 1 ug/mL, respectively, and the susceptibility rate was 92% and 97%, respectively. However, MIC_90 of mecillinam for S. marcescens isolates was >128 ug/mL and most of them were resistant to meclllinam. All Salmonella isolates and 27 of 30 Shigella isolates were susceptible to meclllinam. Conclusion : Mecillinam was active vitro against most Enterobacteriaceae, Salmonella, and Shigella isolates except for S. marcescens. Therefore, mecillinam can be a good alternative agent for treating urinary tract infection and bacterial gastroenteritis.
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