• Korean Journal of Veterinary Research
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• v.40 no.4
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• pp.719-727
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• 2000

A study was conducted to determine the susceptibility of swine to Korean foot-and-mouth disease virus (FMDV; subtype O, isolated from Chungju province) in April, 2ooo. One holstein cow was inoculated intradermolingually with suspension of homogenized tissue from a Korean native cow naturally infected with Korean FMDY. Infected cow was housed with one susceptible cow and one susceptible pig (contact sentinels). Four additional susceptible pigs were housed in the same room but caged separately (non-contacted sentinels). The contacted pig and cow as well as non-contact pigs developed typical clinical signs after 2, 3, and 7 days post exposure, respectively. We compared neutralizing antibody from the animals to FMDV $O_1$ Lombardy, O Taiwan, $O_1$ Campos, and $O_1$ Manisa after 0, 4, 7, 10, 14, 21, 28 days post challenge and post-exposure. The highest viral neutralization titer could be interpreted that serotype O Korea (Chungju isolate) is antigenically more related to $O_1$ Manisa. In addition, immunohistochemistry was used to further characterize Korean FMDV from tissues of infected pigs. Korean FMDV antigen was observed in the tongue, hoof, esophagus, and tonsil tissues of sentinel pigs. These findings suggest that Korean FMD virus isolated from cattle can be rapidly transmitted to pigs both directly and indirectly contrast field observation in which only cattle were clinically ill.

• Korean Journal of Microbiology
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• v.46 no.3
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• pp.303-307
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• 2010

Acinetobacter baumannii 1625, a clinical isolate identified by Vitek and 16S rDNA sequence, showed an extended resistance to most ${\beta}$-lactams including imipenem, kanamycin, gentamicin, tobramycin, and cephalosporins of the third and fourth generations, and produced metallo-${\beta}$-lactamase (MBL) of IMP-1 type which is rare in Korea. The isolate contained a class 1 integron of about 2.5 kb in size and the integron included accA4 (aminoglycoside resistance gene), $bla_{IMP-1}$ (carbapenem resistance gene), and $bla_{OXA-2}$ (extended-spectrum ${\beta}$-lactam resistance gene) gene cassettes in order. The coexistence of IMP-1 type and OXA-2 type ${\beta}$-lactamase gene cassettes in an integron has not been reported in Korea. The transformed integron rendered the E. coli transformant resistant more than eight folds against imipenem, ampicilin, piperacillin, cefazolin, cefoperazone, and aztreonam comparing to the reference strain. This study clearly showed that the extended multi-drug resistance of A. baumannii 1625 was mainly due to the integron.

• The Korean Journal of Mycology
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• v.26 no.1 s.84
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• pp.8-15
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• 1998

The present research was undertaken to investigate the activity of ligninolytic enzymes and the decolorization capability of some dyes with Irpex zonatus BN2, isolated from nature and identified. For the assay of enzyme activities, the isolate did not produce lignin peroxidase (LiP) and veratryl alcohol oxidase (VAO), but laccase and manganese dependent peroxidase (MnP). While the activity for MnP was low $(61.6\;nmol/mg{\cdot}protein)$, its laccase activity was very high $(1185.9\;nmol/mg{\cdot}protein)$. Moreover, laccase had appeared earlier than MnP. When the isolate was incubated with each dye for 10 days, the decolorization rates of azo dyes, such as orange II, orange G, tropaeolin O and congo red were 98.0%, 97.4%, 99.0% and 95.3%, respectively. In case of heterocyclic dyes, eosin Y, toludine blue, methyl blue and azur B were 97.4 %, 98.7%, 99.9% and 94.0% respectively. Finally the results of triphenylmethane dye such as basic fuchsin, malachite green and crystal violet were 98.5%, 95.7% and 99.4%, respectively. The results suggest that laccase of Irpex zonatus BN2 should be played an important role in the decolorization of the dyes.

• Journal of Life Science
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• v.26 no.2
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• pp.220-225
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• 2016

Coagulase-negative staphylococci (CNS) have recently become the bacteria most frequently found in clinical infections. The aim of this study was to investigate the prevalence, antimicrobial susceptibilities, and molecular characteristics of CNS isolates from dental clinic environments in Busan, Korea. One hundred and fifty-four samples were collected from 10 dental clinics and dental hospitals in Busan from December 2014 to January 2015. Species were identified by matrix-assisted laser desorption/ionization–time-of-flight. Antimicrobial susceptibility was determined by disk diffusion methods. A polymerase chain reaction was performed to detect mecA, mupA gene, and SCCmec types. Of the 154 samples, 10(6.5%) isolates were identified as CNS (5 Staphylococcus epidermidis, 2 Staphylococcus capitis, 2 Staphylococcus, and 1 Staphylococcus haemolyticus). Among the 10 isolates, 6 were resistant to penicillin, 5 were resistant to gentamicin, 3 were resistant to tetracycline, and 2 were resistant to cefoxitin and erythromycin. However, clindamycin, ciprofloxacin, teicoplanin, and trimethoprim-sulfamethoxazole resistant isolates were not present. Genes encoding mecA were detected in 4 (2 S. warneri and 2 S. haemolyticus) isolates, and mupA in 1 (S. epidermidis) isolate. One methicillin-resistant CNS (S. warneri) isolate was determined as being of the SCCmec type I. It is concluded that CNS resistant to various antimicrobial agents was widely distributed in dental clinic environments in Korea.

• Biomedical Science Letters
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• v.6 no.3
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• pp.201-208
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• 2000

The vancomycin, one of the family of glycopeptide antibiotics, inhibits the synthesis of bacterial cell wall peptidoglycan and has been widely used against gram-positive bacterial infections, especially for a treatment of methicillin resistant S. aureus infection. However, clinical isolate which was intermediately resistant to vancomycin (Mu50: MIC 8 $\mu\textrm{g}$/ml) was isolated in recent years. In this study we performed vancomycin susceptibility test with the increment method and population analysis with clinical isolates S. aureus. Also we did several kinds of tests with three selected isolates (s129: MIC 7 $\mu\textrm{g}$/ml, s134: MIC 7 $\mu\textrm{g}$/ml, s135: MIC 8 $\mu\textrm{g}$/ml) to find out possible mechanism of vancomycin resistance. As a result, the prevalence of vancomycin resistant S. aureus isolates among S. aureus strains resistant to methicillin was 23.3% (25/107). The vancomycin resistances of isolated strains of S. aureus were between those of Mu5O and Mu3 strains. By PCR analysis, none of the isolates with decreased vancomycin susceptibility contained known vancomycin resistant genes such as vanA, vanB, vanC1, vanC2, and vanH. Major bands of 81 kDa, 58 kDa, 33 kDa, 28 kDa were demonstrable in whole cell lysates by SDS-PAGE from all three isolates as well as reference strains. And especially,45 kDa protein was overproduced in Mu50 strains. Among them increased production of NAD$^{+}$-linked-$_{D}$-lactate dehydrogenase (dnLDH) were detected from one clinical strain (s135) and Mu5O strain. From these data, we suggest that the mechanism of vancomycin resistance in these isolates are distinct from that in enterococci.

• The Journal of the Korean Society for Microbiology
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• v.9 no.1
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• pp.13-18
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• 1974

Blood is one of the most important clinical specimens for the isolation of bacteria. A rapid isolation and a high isolation rate of bacteria are very important in blood culture because bacteremic patients are mostly in grave condition. Various blood culture media which support growth of most fastidious bacteria are available commercially. However, growth of bacteria are frequently delayed because of antibacterial activity of blood. Sodium polyanethol sulfonate(Liquoid) has been reported to inactivate the antibacterial substance and disrupt phagocytic cells. The beneficial effect of SPS is well recognized in the isolation of gram-positive bacteria. However, the effect does not seem to be prominent for gram-negative bacilli isolation mainly due to the rapidity of their growth. It has been experienced with Sal. typhi that the growth is much slower than that of other gram-negative bacilli. For the rapid growth of the organism, use of bile broth has been recommended. Although Sal. typhi is the most frequently isolated organism at present, about one half of total isolates are other organisms and, in case bile broth is used, other media which support growth of these organisms should be used together. Fluid thioglycollate medium(FTM) which is always used in blood culture to isolate anaerobes is inferior to brain heart infusion(BHI) for the isolation of aerobes. This study was done to determine the effect of SPS on the isolation of Sal. typhi from blood. During the Sep. 1973 to Sep. 1974 study period, 2460 blood cultures were made from the Severance hospital patients: BHI and FTM sets 1431 specimens, BHI with SPS(0.05%) and FTM sets 396 specimens, BHI and FTM with SPS sets 359 specimens, BHI and BHI with SPS sets 274 specimens. Mean incubation time required for the macroscopic detection of growth of Sal. typhi were 3.5 days on BHI and 2.7 days on BHI with SPS. The 0.8 day difference was statistically significant. On FTM the mean incubation time was 3.8 days while it was 2.9 days on FTM with SPS. The 0.9 day difference was statistically significant. The result on BHI with and without SPS sets showed faster growth on BRI with SPS in 7 specimens and slower growth in one specimen and the remaining 12 showed growth at the same time. These specimens had mean incubation time of 3.2 days on BHI and 2.3 days on BHI with SPS. The 0.9 day difference was statistically significant. This study indicates beneficial effect of SPS for the rapid isolation of Sal. typhi from clinical blood specimens.

• The Journal of the Korean Society for Microbiology
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• v.35 no.3
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• pp.263-271
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• 2000

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 $bla_{SHV-5}$.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.3
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• pp.125-131
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• 2015

The discovery of activating mutations in EGFR in a subset of lung adenocarcinomas was a major advance in our understanding of lung adenocarcinoma biology, and has led to groundbreaking studies that have demonstrated the efficacy of tyrosine kinase inhibitor therapy. Cytologic specimen procedures have become increasingly popular for obtaining diagnostic material in lung carcinomas. However, frequently the small amount of material or sparseness of tumor cells obtained from cytologic preparations limit the number of specialized studies, such as mutation analysis, that can be performed. In this study we used microdissection to isolate small numbers of tumor cells to assess for EGFR mutations from 76 cytological smear slides of patients with lung adenocarcinomas. We compared our results with previous molecular assays that had been performed on either surgical or cytology specimens as part of the patient's initial clinical work-up. Not only were we able to detect the identical EGFR mutation through the pyrosequencing, but we were also able to consistently detect the mutation from as few as 25 microdissected tumor cells. Furthermore, isolating a purer population of tumor cells resulted in increased sensitivity of mutation detection as we were able to detect mutations from microdissection-enriched cases. Therefore, microdissection can not only significantly increase the number of lung adenocarcinoma patients that can be screened for EGFR mutations, but can also facilitate the use of cytologic samples in the newly emerging field of molecular-based personalized therapies.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.3
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• pp.166-172
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• 2006

Metallo-${\beta}$-lactamase (MBL) can hydrolyze all ${\beta}$-lactams except monobactams and frequently coexists with various antibiotic resistance genes such as aminoglycoside resistance, sulfonamide resistance gene, etc. Therefore, the effective antibiotics against infections by these bacteria are markedly limited or can't even be found. We tried to search in-vitro antimicrobial combinations with synergistic effects for a VIM-2 type MBL producing Pseudomonas aeruginosa, isolated from clinical specimen. On the selection of antibiotic combinations with synergistic effects, we performed a one disk synergy test, modified Pestel's method, in agar without aztreonam (AZT). The bacteriostatic synergistic effects of this tests were scored as $S_1$ (by susceptibility pattern in agar without antibiotics), $S_2$ (by the change of susceptibility in agar with or without antibiotics) and $S_3$ ($S_1$ + $S_2$) and was classified into weak (1 point), moderate (2 points) and strong (3 points) by $S_3$ score. Subsequently, we carried out the time-killing curve for the antibiotic combinations with the strong synergistic bacteriostatic effect. One VIM-2 type MBL producing P. aeruginosa confirmed by the PCR showed all resistance against all ${\beta}$-lactams except AZT, aminoglycoside and ciprofloxacin. In the one disk synergy test, this isolate showed a strong bacteriostatic synergistic effect for the antibiotic combination of AZT and piperacillin-tazobactam (PIP-TZP) or AZT and amikacin (AN). On the time-killing curve after six hours of incubation, the colony forming units (CFUs/mL) of this bacteria in the medium broth with both combination antibiotics were decreased to 1/18.7, 1/17.1 of the least CFUs of each single antibiotics. The triple antibiotic combination therapy including AZT, PIP-TZP and AN was shown to be significantly synergistic after 8 hrs of exposure. In a VIM-2 MBL producing P. aeruginosa with susceptibility for AZT, the triple antibiotic combination therapy including AZT, PIP-TZP and AN may be considered as an alternative antibiotics modality against the infection by some MBL type. But the antimicrobial combination therapy for many more MBL producing isolates is essential to know as soon as possible for the selection of effective treatment against the infection by this bacteria.

• Tuberculosis and Respiratory Diseases
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• v.58 no.5
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• pp.452-458
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• 2005

Background : In Korea, polymerase chain reaction (PCR) test for M. tuberculosis has been used for the diagnosis of acid-fast bacilli (AFB) smear-negative tuberculosis in order to increase diagnostic sensitivity. However, there have been no data dealing with the clinical utility of PCR in AFB smear-positive patients to differentiate between M. tuberculosis and nontuberculous mycobacteria. Method : We retrospectively analyzed the PCR test results which have been performed in patients who had AFB smear-positive sputum but had ambiguous clinical manifestations of active tuberculosis. PCR test was done using $AMPLICOR^{\hat{a}}$ M. tuberculosis kit. The sensitivity, specificity, and positive and negative predictive values of the PCR test were calculated based on culture and final clinical diagnosis result. Results : Fifty-six consecutive patients (62 PCR tests) were included in the study. Active tuberculosis was diagnosed in 23 patients (41.0%), while 9 patients had NTM infection (16.0%). The sensitivity, specificity, positive- and negative-predictive value of PCR test were 88.8%, 86.8%, 76.1% and 94.3%, respectively, according to the culture result. In comparison, they were 91.3%, 100%, 100%, 94.3%, respectively, according to the final clinical diagnosis. All 15 patients with NTM isolates, including 6 patients who had other lung diseases but expectorated NTM isolate, were negative for PCR test. Conclusion : Even though tuberculosis is still prevalent in Korea, PCR test is useful to differentiate between M. tuberculosis and NTM in patients with AFB-smear positive sputum but with ambiguous clinical manifestations of active tuberculosis.