• Title/Summary/Keyword: Frequency of resistance

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Integrated Rotary Genetic Analysis Microsystem for Influenza A Virus Detection

  • Jung, Jae Hwan;Park, Byung Hyun;Choi, Seok Jin;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.88-89
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    • 2013
  • A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

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FES Exercise Program for Independent Paraplegic Walking (하반신 마비환자의 FES 독립보행을 위한 근육 강화 프로그램)

  • Khang, Seon-Hwa;Khang, Gon;Choi, Hyun-Joo;Kim, Jong-Moon;Chong, Soon-Yeol;Chung, Jin-Sang
    • Journal of Biomedical Engineering Research
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    • v.19 no.1
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    • pp.69-80
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    • 1998
  • This research was designed to investigate how the exercise program affects paraplegic standing and walking employing functional electrical stimulation(FES). Emphasis was also given to fatigue of major lower extremity muscles induced by different types of electrical stimulation. We applied continuous and intermittent rectangular pulse trains to quadriceps of 10 normal subjects and 4 complete paraplegic patients. The frequencies were 20Hz and 80Hz, and the knee angle was fixed at 90$^{\circ}$and 150$^{\circ}$to investigate how muscle fatigue is related to muscle length. The knee extensor torque was measured and monitored. We have been training quadriceps and gastrocnemius of a male paraplegic patient by means of electrical stimulation for the past two year. FES standing was initiated when the knee extensors became strong enough to support the body weight, and then the patient started FES walking utilizing parallel bars and a walker. We used an 8-channel constant-voltage stimulator and surface electrodes. The experimental results indicated that paralyzed muscles fatigued rapidly around the optimal length contrary to normal muscles and confirmed that low frequency and intermittent stimulation delayed fatigue. Our exercise program increased muscle force by approximately 10 folds and decreased the fatigue index to half of the initial value. In addition, the exercise enabled the patient to voluntarily lift each leg up to 10cm, which was of great help to the swing phase of FES walking. Both muscle force and resistance to fatigue were significantly enhanced right after the exercise was applied every day instead of 6 days a week. Up to date, the patient can walk for more than two and half minutes at 10m/min while controlling the on/off time of the stimulator by pushing the toggle switch attached to the walker handle.

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Trends in Serotype Distribution of Clinical Isolates of Streptococcus pneumoniae: A Single Center Experience from 2001 to 2006 (임상검체에서 분리된 폐구균 혈청형의 변동추이)

  • Lee, Taek Jin;Chun, Jin Kyong;Choi, Kyoung Min;Yong, Dong Eun;Lee, Kyoung Won;Kim, Dong Soo
    • Pediatric Infection and Vaccine
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    • v.13 no.2
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    • pp.115-123
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    • 2006
  • Purpose : We investigated whether there had been any change in the epidemiology of Streptococcus pneumoniae in Korea before and after introduction of heptavalent pneumococcal conjugate vaccine(PCV7). Methods : Between September 2001 and August 2006, clinical isolates were collected from patients with pneumococcal infection in Severance Hospital, Seoul, Korea. We analyzed trends in serotype distribution and antibiotic resistance before and after the introduction of PCV7. Results : There were 363 strains of Streptococcus pneumoniae isolated from clinical specimens; 143 before and 220 after PCV7 introduction. The predominant serotypes, in order of decreasing frequency, were 19F, 19A, 23F, 6B, 6A, 3, 9V, 14, 11A, 4, 29, and 18C; 152 isolates(41.9%) belonged to types included in PCV7. The proportion of clinical isolates that were nonsusceptible to penicillin increased from 58.8% in 2001 to 83.6% in 2006(P=0.046). There was no significant diminution in pneumococcal infection caused by vaccine serotypes after the introduction of PCV7. In children younger than age 2 years, the proportion of clinical isolates that were vaccine serotypes was higher than in persons older than age 15 years (59.3% vs 37.8%, P=0.004). Conclusions : There was no significant diminution in pneumococcal infection caused by vaccine serotypes after the introduction of PCV7, therefore more universal pneumococcal immunization program is recommended especially for children younger than age 2 years.

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The Effect of Water Conveyance of Boryeong Dam on Structural Changes of Benthic Macroinvertebrates Community (보령댐 도수가 저서성 대형무척추동물 군집구조 변화에 미치는 영향)

  • An, Chae-Hui;Han, Jung-Soo;Choi, Jun-Kil;Lee, Hwang-Goo
    • Korean Journal of Environment and Ecology
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    • v.32 no.4
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    • pp.381-391
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    • 2018
  • The purpose of this study is to investigate the influence of artificial disturbances occurring during water conveyance frequency on benthic macro-invertebrate by comparing and analyzing data before and after the construction of Boryeong dam in Boryeong-si, Chungcheongnam-do. We conducted the survey eight times in four points from April 2016 to September 2017. A total of 13,447 individuals from 125 species, 70 families, 19 orders, 7 classes, and 4 phyla were collected. The result of the community analysis showed that the dominant index was 0.55 (${\pm}0.07$) to 0.47 (${\pm}0.08$), diversity index was 2.06 (${\pm}0.26$) to 2.23 (${\pm}0.28$), evenness index was 0.69 (${\pm}0.05$) to 0.70 (${\pm}0.04$), and richness index was 3.43 (${\pm}0.72$) to 4.03 (${\pm}0.91$). After conveyance, the dominance decreased while the diversity, evenness, and richness increased. The result of the functional group analysis showed the portion of scrapers, filtering-collectors increased while that of shredders, gathering-collectors decreased in the functional feeding group and that the proportion of climbers and clingers increased while that of burrowers and sprawlers decreased in the habitat orientation group. The result of analysis of community stability showed that St. 1 and St. 2 decreased resistance and resilience after conveyance and new species appeared in the III characteristics group while St. 3 and St. 4 did not show much difference. The result of multidimensional scaling analysis showed that the variation of similarity was the highest at St. 1 after conveyance, and the variation of similarity at St. 4 was the lowest.

Studies on the Taxonomical Characters in Populus tomentiglandulosa and P. glandulosa (Populus tomentiglandulosa와 P. glandulosa의 분류학적(分類學的) 연구(硏究))

  • Kim, Sam Sik;Kim, Chung Suk;Noda, Shozo
    • Journal of Korean Society of Forest Science
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    • v.51 no.1
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    • pp.1-21
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    • 1981
  • Populus glandulosa and Populus tomentiglandulosa, which were known to be natural hybrids, were examined for morphological, physiological and karyological traits to illucidate its hybridity and taxonomical importance. The results abtained were as follows; 1. Survival rate in rooting of cuttings and grafting was different between the hybrids and their rooting abilities showed incomplete dominance. 2. Their leaf openings showed incomplete dominance. The leaf longevities of P. alba ${\times}$ glandulosa and P. tomentiglandulosa were stronger than the other hybrids. 3. There were differences in resistance to toxicity of $KClO_3$ between the hybrids. 4. Many external leaf characters of the hybrids also showed incomplete dominance. P. tomentiglandulosa was similar in those characters to P. alba ${\times}$ glandulosa while P. glandulosa was similar to hybrids crossed, reciprocally crossed or back-crossed between P. davidiana and P. alba. 5. Their numbers of male flower showed incomplete dominance or hybrid vigor. The numbers of P. tomentiglandulosa were similar to thosa of P. alba ${\times}$ glandulosa while those of P. glandulosa to those of P. alba ${\times}$ davidiana or P. davidiana ${\times}$ alba. 6. Morphology and band color of male catkin bract showed incomplete dominance. Those of P. glandulosa were similar to those of P. alba ${\times}$ davidiana while those of P. tomentiglandulosa to those of. P. alba ${\times}$ glandulosa. 7. There were differences in vascular bundle number and arrangement of petiole between the hybrids. 8. Differences in the anatomical traits of stem did not exist between the hybrids but those in wood fiber size existed. 9. The chromosomes of artificial hybrids, P. glandulosa and P. tomentiglandulosa showed irregular behavior in metaphase I and II. 10. All hybrids including P. glandulosa and P. tomentiglandulosa showed small number of P.M.C. with 19 II but many univalent chromosomes were exhibited in metaphase I. 11. All hybrids including P. glandulosa and P. tomentiglandulosa showed a little abnormal nuclear plates as laggard chromosome and chromosome bridge in anaphase I and II. 12. The frequency of pollen tetrad and fertile pollen was low in most of the hybrids including P. glandulosa and P. tomentiglandulosa.

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Evaluation of the Cell-Mediated Immunity in Treatment Failure Pulmonary Tuberculosis (치료실패 폐결핵 환자의 세포성면역반응에 관한 연구)

  • Park, Jeong-Kyu;Park, Jang-Seo;Kim, Hwa-Jung;Jo, Eun-Gyeong;Min, Dul-Lel;Lim, Jae-Hyun;Suhr, Ji-Won;Paik, Tae-Hyun
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.1
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    • pp.13-25
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    • 1999
  • Background: Ineffective cell-mediated immune response in human tuberculosis is associated with a depressed Thl cytokine response and reduced production of IFN-$\gamma$. Most persons infected with Mycobacterium tuberculosis are healthy tuberculin reactors with protective immunity, but a minority with ineffective immunity develop extensive pulmonary tuberculosis. The cell-mediated immune response is an important aspect of host resistance to mycobacterial infection and is believed to be tightly regulated by a balance between Th1 cytokines including IFN-$\gamma$, IL-12, IL-18, regulated on activation, normal T cell expressed and secreted (RANTES) and Th2 counterparts such as IL-4, monocyte chemoattractant protein-l (MCP-l). Methods: Proliferation and mRNA expression of IFN-$\gamma$, RANTES and MCP-l by RT-PCR in peripheral blood mononuclear cells (PBMCs) in response to in vitro stimulation with mycobacterial antigens were compared in pulmonary tuberculosis patients with cured and treatment failure and in tuberculin-positive and tuberculin-negative healthy subjects. Results: Defective proliferative responsiveness to aqueous TSP antigen was involved with treatment failure tuberculosis patients. Aqueous TSP antigen-induced IFN-$\gamma$ and RANTES mRNA expression was decreased in treatment failure tuberculosis patients compared with healthy tuberculin reactors and cured tuberculosis patients (23.1 % versus 90.0% for IFN-$\gamma$ and 46.2% versus 70.0% versus 46.2% for RANTES). The frequency of MCP-l mRNA expression to aqueous TSP antigen in treatment failure tuberculosis patients was greater than in healthy tuberculin reactors and cured tuberculosis patients (76.9% versus 40.0%). Conclusion: The increasing expression of MCP-1 mRNA in response to aqueous TSP antigen might be predicted to favor Th1 responses and restricted Th1 responses in treatment failure of pulmonary tuberculosis.

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Socio-medical Study on Gonorrhoea with Special References of Prevalence, PPNG and Antibiotic Resistance (임질환자(淋疾患者)에 관(關)한 사회의학적(社會醫學的) 연구(硏究) -유병률(有病率), PPNG 및 항균제내성(抗菌劑耐性)을 중심(中心)으로-)

  • Lee, Seong-Ho;Hwang, In-Dam;Park, Young-Soo;Koh, Dai-Ha
    • Journal of Preventive Medicine and Public Health
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    • v.16 no.1
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    • pp.41-50
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    • 1983
  • The incidence of sexually transmitted disease, especially gonorrhoea has risen despite the progress in its diagnosis and treatment. For the effective control programs of sexually transmitted disease, it should be required socio-medical approaches. A study on gonorrhoea and penicillinase-producing Neisseria gonorrhoeae(PPNG) was conducted in Jeonju and Kunsan area from March, 1982 through August, 1982. The 221 entertrainers were studied in order to determine the prevalences of gonorhoea and PPNG. Socio-demographic informations of the entertainers were obtained by interviewing them. Gonococci were cultured on Thayer-Martin enrichment media for isolation, and PPNG was confirmed using beta-lactamase reagent ($PADAC^{tm}$ Beta-Lactamase Test Strips, Galbiochem-Behring). The results of the study are summarized as follows; 1. The average age of the entertainers studied was $26.1{\pm}4.7$ years. 2. The average years of working in entertaining business was $2.4{\pm}1.4$ years, and the average income per month was $239,592{\pm}90,480$ won. On the education level, 70.6% of the entertainers were middle or high school graduates 3. 47.5% of the entertainers were using contraceptives. 90.5% have experienced artificial abortion. 4. 37(16.7%) out of 221 entertainers were revealed to gonorrhoea, and 13 (35.1%) of gonorrhoea patients were by PPNG. 5. The prevalence rates of gonorrhoea and the proportion of PPNG by age were not significant statistically. Meanwhile, the colelations between the rates of gonorrhoea and education, frequency of love-making with customers and type of sexual partner were highly significant statistically. 6. 37 strains of gonococci isolated were almostly resistant to several antimicrobial agents, especially amikacin, clindamycin and chloramphenicol. Furthermore PPNG strains were completely resistant to not only above drugs but also penicillin.

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Distribution Status of Invasive Alien Species (Procambarus clarkii (Girard, 1852)) Using Biomonitoring with Environmental DNA in South Korea (생물모니터링 및 환경유전자(eDNA)를 이용한 침입외래종 미국가재(Procambarus clarkii)의 한국분포 현황)

  • Jung, Sang Woo;Lee, Jae-Ha;Kawai, Tadashi;Kim, Phil-Jae;Kim, SuHwan
    • Korean Journal of Environment and Ecology
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    • v.36 no.4
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    • pp.368-380
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    • 2022
  • Biomonitoring of an invasive alien crayfish species, Procambarus clarkii (Girard, 1852), was performed from February to October 2021, along with environmental DNA analysis, at five locations including Wanju-gun, Hampyeong-gun, Naju-si, Gurye-gun, and Cheongju-si. For the investigation, an umbrella-shaped trap for adults and a hand net for young crayfish were used, and 8 to 10L of freshwater was collected for eDNA analysis. The current status and past distributional records of crayfish in Korea were analyzed along with benthic macroinvertebrates at each survey site. As a result of the investigation, a total of 122 individuals were identified, and Hampyeong-gun recorded the largest number of populations with 59 individuals (48.36%) and the highest environmental DNA (eDNA). The frequency of appearance of P. clarkii was highest in May. The ratio of females to males was 21:5, and the body size was 72.2±21.1mm for female, 80.5±15.6mm for male, and 25.3±9.8mm for young crayfish. P. clarkii introduced into Korea is mainly spreading in the southwest region and it has not been observed in Seoul where there had been a record of appearance in the past. No external symbiosis (Branchiobdellida) of P. clarkii has been identified, and P. clarkii that has appeared in Korea was presumed to be imported from Japan. There are more than eight kinds of exotic crayfish distributed in Korea, and among them, the marbled crayfish (P. virginalis) was identified as a harmful species to the ecosystem of Korea by the Ministry of Environment in 2021. The identified species of benthic macroinvertebrates inhabiting the survey area where P. clarkii has appeared were 69 belonging to 39 families, 15 orders, five classes, and three phyla. Among them, Odonata were the most abundant (16 spp.; 24.62%), followed by Coleoptera (11 spp.; 16.92%) and Hemiptera (11 spp.; 16.92%). In the survey area, one Korean endemic species (Rhoenanthus coreanus), one species (Helophorus auriculatus) classified as Near Threatened (NT) on the Korean Red List, and six species of Crustaceans appeared. In the functional feeding group, the predators appeared predominantly while in habitat oriented group, the climbers appeared to be abundant. It was confirmed that P. clarkii in the survey area prefers an area rich in aquatic vegetation with waterside vegetation, and has high resistance to turbid water quality. The omnivore invader P. clarkii is expected to maintain a competitive relationship with carnivorous benthic macroinvertebrates that are predominant in the same species, and is expected to continue to generate ecosystem disturbance along the food chains.

Effect of Inhaled Nitric Oxide on Hemodynamics, Gas Exchange and Pulmonary Inflammation in Newborn Piglets with Escherichia coli Induced Septic Lungs (패혈성 폐가 유도된 신생자돈에서 외인성 Nitric Oxide 흡입의 혈역학, 가스교환 및 폐 염증에 대한 효과)

  • Chang, Yun Sil;Ko, Sun Young;Park, Won Soon
    • Clinical and Experimental Pediatrics
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    • v.46 no.8
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    • pp.777-783
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    • 2003
  • Purpose : The aim of this study was to evaluate the effect of inhaled nitric oxide(iNO) on gas exchange, hemodynamics and pulmonary inflammation in newborn piglets with E. coli induced septic lung. Methods : Twenty three instrumented and ventilated piglets were randomized into three groups : CON(n=6), PCON(n=9), and PNO(n=8). In the piglets of the PCON and PNO groups, E. coli septic lung was induced by endotracheal instillation of E. coli. Ten ppm iNO was given continuously in the PNO group after endotracheal instillation of E. coli. All animals were mechanically ventilated for six hour with a peak inspiratory pressure of 30 $cmH_2O$, frequency of 25 breaths/min, $FiO_2$ 1.0 and a positive end-expiratory pressure of 4 $cmH_2O$. All measurements were made at one hour intervals during the experiment. At the end of the experiment, lung tissue was harvested for the analysis of myeloperoxidase activity, indicative of lung inflammation. Results : All piglets with pulmonary instillation of E. coli developed E. coli sepsis. Piglets in the PCON group developed progresseve pulmonry hypertension, hypoxemia and hypercarbia compared to the CON group due to increased pulmonary vascular resistance, intrapulmonary shunt fraction and physiologic dead space fraction. iNO did not reverse pulmonary hypertension in the PNO group. However iNO significantly improved oxygenation, which was attributed to marked improvement of venous admixture and partial attenuation of increase in dead space fraction. Increased myeloperoxidase activity in PCON compared to CON was significantly attenuated in PNO. Conclusion : iNO improves oxygenation and lung inflammation in newborn piglets with E. coli induced septic lung.

Genotypic Investigation of Multidrug-Resistant Pseudomonas aeruginosa from Clinical Isolates in Korea, 2010 (2010년도 국내 임상에서 분리한 다제내성 녹농균의 유전자형 조사)

  • Kim, Min Ji;Cha, Min Kyeong;Lee, Do Kyung;Kang, Ju Yeon;Park, Jae Eun;Kim, Young Hee;Park, Il Ho;Shin, Hea Soon;Ha, Nam Joo
    • Korean Journal of Microbiology
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    • v.48 no.4
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    • pp.240-245
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    • 2012
  • Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium that causes serious infection, particularly in immunocompromised patients. Also, P. aeruginosa possessing carbapenem-resistant metallo-${\beta}$-lactamases (MBL) has been reported with increasing frequency in Korea. We therefore analyzed the level of multidrug-resistant clinical P. aeruginosa isolated from a secondary hospital in Korea in 2010. A total of 92 isolates of P. aeruginosa were collected from Sahmyook Medical Center in 2010. Susceptibility to antimicrobial agents was determined by analysis of the minimum inhibitory concentration test; the inhibitor-potentiated disk diffusion (IPD) test was performed for MBL detection. RAPD-PCR was used for genotyping to rapidly characterize P. aeruginosa strains isolated from clinical patients. The percentages of non-susceptible isolates were as follows: 40.2% to ceftazidime, 58.7% to meropenem, 56.5% to gentamicin, 46.7% to tobramycin, 62.0% to ciprofloxacin and 97.8% to chloramphenicol. The 29 multidrug-resistant strains were screened by the IPD test: of the 21 PCR-positive isolates, 19 were IPM-1 producers and 2 were VIM-2 producers. Among the 19 IMP-1-producing P. aeruginosa isolates, 16 isolates showed similar patterns, and three different banding patterns were observed. The proportion of IMP-1-producing multidrug-resistant P. aeruginosa from clinical isolates steadily increased in this secondary hospital in Korea in 2010. This study provides information about the antimicrobial-resistant patterns and genotype of multidrug-resistant P. aeruginosa isolated from clinical isolates in Korea, 2010.