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혈소판 제제에서 세균 오염 검출을 위한 방법의 민감도 비교
이혁민,박윤희,임환섭 대한수혈학회 2009 大韓輸血學會誌 Vol.20 No.1
Background: The demand for platelet concentrates has increased for patients with hemato-oncologic diseases as well as for patients with chronic diseases. As platelet concentrates are preserved at 22∼24oC, the chance of bacterial contamination exposure is increased, which can cause fatal outcomes. We evaluated various methods for detecting bacterial contamination in platelet concentrates. Methods: 0.5 MacFarland standard solutions were prepared using the Staphylococcus aureus ATCC 25923 & Escherichia coli ATCC25922 strains. The platelet concentrates were inoculated with various concentrations (101∼105 CFU/mL) of bacteria and then gram staining, plate culture, broth culture and 16s RNA were used to detect bacteria. Results: The gram stain method was unable to detect bacteria concentrations less than 104 CFU/mL. The plate culture method detected bacterial growth concentrations up to 103 CFU/mL, but only 1 specimen of S. aureus was detected at the lowest concentration of 101 CFU/mL. The broth culture method detected 102 CFU/mL concentrations except for samples from S. aureus and E. coli strains. Among the 101 CFU/mL lowest concentrations, bacterial growth detected 3 samples from S. aureus and 2 samples from E. coli. For the broth culture method, detection of bacterial growth up to 101 CFU/mL took 58.9 hours, it took 57.5 hours for S. aureus and E. coli respectively, and it took 43.9 hours and 49.0 hours for 102 CFU/mL concentrations of S. aureus and E. coli, respectively. The PCR method showed all positive results except for 1 specimen of E. coli. Conclusion: The broth culture method showed similar sensitivity to PCR except for the 43.9∼58.9 hours of an incubation period to show positive results. Overall, the PCR method was most sensitive and rapid method for detecting bacterial contamination in platelet concentrates. 배경: 혈소판 제제는 혈액암 환자를 포함한 만성 환자에서 근래에 사용이 증가하고 있다. 혈소판 제제는 실온에 보관하므로 세균 오염에 민감하며 위중한 결과를 초래할 수 있어, 이에 대한 주의가 필요하다. 본 연구에서는 이러한 혈소판 제제의 세균 오염을 신속하게 정확하게 진단할 수 있는 미생물학적 검사 방법을 평가하고자 하였다. 방법: Staphylococcus aureus ATCC 25923 및 Escherichia coli ATCC 25922 균주를 이용하여 0.5 McFarland 표준 균액을 제조하였다. 제조한 표준 균액으로 101∼105 CFU/mL의 농도를 갖는 혈소판 액을 제조하고, 각 농도에서 그람 염색, 평판 배지 배양, 액체 배지 배양(BacT/ALERT 3D system, bioMerieux Inc., Marcy E'toille, France) 및 16s rRNA PCR을 시행하였다. 결과: 그람 염색은 104 CFU/mL 보다 낮은 농도에서는 균을 검출할 수 없었다. 평판 배지 배양법에서는 103 CFU/mL 농도까지는 모든 균액에서 균의 성장을 확인할 수 있었으나 가장 낮은 101 CFU/mL 농도에서는 S. aureus 1 검체에서만 균을 확인할 수 있었다. 액체 배지 배양은 102 CFU/mL의 농도까지는 S. aureus 및 E. coli 각각 1검체를 제외한 모든 검체에서 균을 확인할 수 있었으며, 가장 낮은 101 CFU/mL 농도에서도 S. aureus 3 검체 및 E. coli 2 검체에서 균을 확인할 수 있었다. 액체 배지 배양에서 균의 증식이 확인되기까지 101 CFU/mL 농도에서 평균적으로 S. aureus 58.9 시간 및 E. coli 57.5시간이 소요되었으며, 102 CFU/mL의 농도에서는 S. aureus 43.9시간 및 E. coli 49.0시간이 소요되었다. 분자 유전학적인 검사법인 PCR에서는 E. coli 1 검체를 제외한 모든 검체에서 양성 반응을 보였다. 결론: 액체 배지 배양법은 PCR법과 비슷한 정도의 민감도를 보였으나 검사 양성까지 걸리는 시간이 43.9∼58.9시간으로 길었으며, PCR 법은 민감하고 신속하게 혈소판 제제의 오염을 확인할 수 있을 것으로 판단되었다.
2004년도 국내 12개 병원에서 분리된 주요 세균의 항균제 내성율
이혁민,용동은,이경원,홍성근,김의종,정석훈,박연준,최태열,어영,신종희,이위교,이종욱,안지영,이성희,우건조 대한임상미생물학회 2005 Annals of clinical microbiology Vol.8 No.1
Background: A rapid increase in antimicrobial-resistant bacteria has become a serious problem in many countries including Korea, but the rate and pattern of antimicrobial resistance may vary significantly depending on countries and even on hospitals. The aim of this study was to determine the nationwide prevalence of resistance among frequently isolated bacterial pathogens in Korea. Methods: Routine susceptibility data for medically important bacterial pathogens from 12 university hospital and general hospital laboratories in Korea were analysed by patient group. These pathogens had been isolated during the period from April to November in 2004. Results: The proportion of methicillin-resistant Staphylococcus aureus (MRSA) was 67%. Vancomycin-resistance rate of Enterococcus faecalis was 1% and that of E. faecium was 20%. The resistance rates of Streptococcus pneumoniae to penicillin and Haemophilus influenzae to ampicillin were 70% and 54%, respectively. The resistant rates of Escherichia coli and Klebsiella pneumoniaewere 7-10% and 26-31% to the 3rd generation cephalosporin, respectively. The resistance rates to 3rd generation cephalosporin were 22-30% in Citrobacter freundii, 35-44% in Enterobacter cloacaeand 15-22 % in Serratia marcescens. Imipenem resistance rates of Pseudomonas aeruginosaand Acinetobacter baumannii were 26% and 17%. Cotrimoxazole and levofloxacin resistance rates of Stenotrophomonas maltophiliawere 46% and 44%, respectively. Conclusion: Antimicrobial resistance rates of clinically important pathogens in Korea were still high and were generally higher among the bacteria isolated from the intensive care unit patients. Strict infection control and continuous nationwide surveillance program will be required to manage the antimicrobial resistance problem.
이혁민,Kyung Ho Roh,홍성근,신희봉,정석훈,송원근,어영,용동은,이경원 대한진단검사의학회 2016 Annals of Laboratory Medicine Vol.36 No.2
Background: Extensively drug-resistant (XDR) Pseudomonas aeruginosa and Acinetobacter baumannii are a threat to hospitalized patients. We evaluated the effects of antimicrobial combinations on XDR P. aeruginosa and A. baumannii isolates. Methods: P. aeruginosa and A. baumannii isolates, which were resistant to all antibiotics except colistin (CL), were collected from eight hospitals in Korea. Genes encoding metallo-β-lactamases (MBLs) and OXA carbapenemases were detected by PCR in eight P. aeruginosa and 30 A. baumannii isolates. In vitro synergy of antimicrobial combinations was tested by using the checkerboard method. Results: Minimum inhibitory concentrations of β-lactams, aminoglycosides, and fluoroquinolones were very high, while that of CL was low for majority of XDR P. aeruginosa and A. baumannii isolates. Antimicrobial combinations including Imipenem (IPM)-CL, ceftazidime (CAZ)-CL, and rifampin (RIF)-CL exerted only additive/indifferent effects on majority of XDR P. aeruginosa isolates. Proportions of XDR A. baumannii isolates that showed synergistic and additive/indifferent inhibition after treatment with antimicrobial combinations used are as follows: IPM-ampicillin-sulbactam (AMS), 17% and 80% isolates, respectively; IPM-rifampin (RIF), 13% and 81% isolates, respectively; IPM-CL, 13% and 87% isolates, respectively; and RIF-COL, 20% and 73% isolates, respectively. Significant proportion (19%) of XDR P. aeruginosa isolates produced MBLs, and majority (82%) of A. baumannii isolates produced either MBLs or OXA-23. Conclusions: Our results suggest that combinations of IPM-AMS, IPM-RIF, IPM-CL, and RIF-CL are more useful than individual drugs for treating 13-20% of XDR A. baumannii infections.
Antimicrobial Resistance of Neisseria gonorrhoeae Isolated in Korea
이혁민,이경원,정윤섭 대한미생물학회 2012 Journal of Bacteriology and Virology Vol.42 No.1
Neisseria gonorrhoeae is the causative agent of gonorrhea, one of the most important sexually transmitted diseases. The incidence of gonorrhea is still prevalent and about 50,000 new cases have been reported annually during the late 2000s in Korea. The antimicrobial resistance of N. gonorrhoeae is very prevalent and most isolates are multi-drug resistant to penicillin G, tetracycline, and fluoroquinolones. The incidence of penicillinase-producing N. gonorrhoeae (PPNG) decreased significantly, but high-level tetracycline-resistant N. gonorrhoeae (TRNG) increased recently. The minimum inhibitory concentrations (MICs) of ceftriaxone were within the susceptible range for all isolates, but MIC creep has been apparent and one cefixime-nonsusceptible isolate (0.5 μg/ml) was found. Spectinomycin-resistant isolates remain rare, but caution should be required when dealing with gonococcal pharyngitis.