• Title/Summary/Keyword: clinical microbiology

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Efficient Generation of BLCL Expressing Foreign Antigen as Antigen-presenting Cells with Recombinant Retroviruses

  • Hyun-Il Cho;Soon-Young Pail;Il-Hoan OH;Kyun-Jung Ahn;Dong-Wook Kim
    • Journal of Microbiology
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    • v.39 no.4
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    • pp.300-304
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    • 2001
  • Epstein-Barr Virus(EBV)-transformed lymphoblastoid B cell lines, BLCL which expresse antigens, are potential antigen-presenting cells(APCs) for the induction of CTL in vitro. However transfection of BLCLs with subsequent selection by antibiotics is notoriously difficult because plating efficiencies of BLCLsare reported to be 1% or less. To generated stable transfectants of BLCLs we produced high titers of retroviruess encoding pp 65 antigen of human cytomegalovirus of foreign antigens and trans-duced them of BLCLs. The pp 65 gene was cloned into the retroviral vector pLXSN. The recombinant retroviral vector was transfected to ecotropic packaging cell line, CP&E86, and this polyclonal recom-binant retrovirus was transduced to PA317 that is amphotropic pakaging cell line. The titers of colned PA317 amphotropic retroviruses ranged from 5 to $\times$10$^{6}$ colony forming units (CFU)per ml (CFU/ml) We performed three rounds of consecutive transductions to BLCLs in order to improve the clon-ing effieiencies. The expression of recombinant HCMV-pp65 antigen was more than 20% after the final transduction. THe third-transduced BLCLs were easily selected in optimal concentration of G418. BLCLs expressing foreign antigens could be used as target cells for CTL assay and/or as APCs for induction of in vitro CTL responses specific for viral and tumor antigens.

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Serological grouping of $\beta$-hemolytic streptococci by a coagglutination technique (Coagglutination에 의한 $\beta$-용혈성 연쇄구균의 혈청군 동정)

  • Chong, Yun-Sop;Yoon, Yang-Sook;Kim, Yoon-Chung;Lee, Sam-Uel Y.;Kim, Sung-Kwang;Lee, Byung-Soo;Kim, Joo-Deuk
    • The Journal of the Korean Society for Microbiology
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    • v.14 no.1
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    • pp.11-15
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    • 1979
  • Identification of group A $\beta$-phemolytic streptococci is very important to provide an appropriate preventive measure of possible rheumatic fever and acute glomerulonephritis. For such purpose bacitracin susceptibility of streptococci because of its simplity has been most widely used despite of its occasional faulty results. Recently, a coagglutination technique was advocated using streptococcal group specific antibodies adsorbed to protein A-containing staphylococci. This study was conducted to evaluate the coagglutination technique using reagents prepared by ourselves. The specificity, reproducibility and stability were ascertained and the following results were obtained. 1. The identification by coagglutination technique using our own reagent gave the same results compared with the Lancefield precipitation technique. The result also agreed with the Phadebact grouping. 2. There were no variation in group A and B identification due to lot difference. However, there were a few discrepant results in group C and G identification which was conducted in different days with different lots of our reagent. 3. The stability of our reagents was less satisfactory compared to the commercial product. An effort to improve the stability was considered necessary. 4. For coagglutination, it was found convenient to use supernatant of Todd-Hewitt broth incubated for 24 hours. Both parafin-ringed slide glass and RPR card gave comparable results and the former could be used when the latter is not available.

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Production of Monoclonal Antibody to Chlamydia Trachomatis (Chlamydia trachomatis 진단에 유용한 단세포군 항체 생산에 관한 연구)

  • Choi, Tae-Yeal;Kim, Think-You;Kim, Choon-Won;Kim, Ki-Hong;Hwang, Eung-Soo;Cha, Chang-Yong;Kim, Kwang-Hyuk
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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    • pp.197-208
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    • 1987
  • Chlamydia trachomatis has now shown that this interesting intracellular parasite is a cause of nongonococcal urethritis, infantile pneumonia, pelvic inflammatory disease and epididymitis, in addition to lymphogranuloma venerum and inclusion conjunctivitis. There are several diagnostic methods for C. trachomatis, but the method using monoclonal antibody is the most sensitive and specific. The hybride cell were prepared by fusion of myeloma cell($P_3X_{63}\;Ag_8{\cdot}V_{653}$) of mouse and lymphocyte of mouse(BALB/c) that were immunized with formalin killed C. trachomatis serotype D. The cell mixtures after fusion were dispensed into 640 wells of the 96 well culture plates and continuously cultured in HAT medium for 2 weeks. The supernatants of culture media in 83(13%) wells were reacted with C. trachomatis, which were determined by enzyme-linked immunosorbent assay in 96 well microplate. The clones that secreted antibody to C. trachomatis were cloned by limiting dilution. Only six monoclones secreted antibody to C. trachomatis. The antibody titer of ascitic fluid that collected from same BALB/c mice bearing hybridoma cells was above 1:100,000. These monoclonal antibodies that were IgG reacted with elementary and reticulate bodies of all serotypes(Ba, D, E, F, G, H, J and LGV type-I) using ELISA and indirect immunofluorescence stain, but there were no cross reaction with other bacteria(coagulase negative Staphylococcus, Proteus and E. coli). We concluded these six monoclones secreted the same monoclonal antibody to C. trachomatis. The sensitivity and specificity of the monoclonal antibody compared with Microtrak(confirmatory test of C. trachomatis, Syva) was 100%, respectively.

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Introduction of Clinical and Laboratory Standards Institute Antibiotic Susceptibility Testing Subcommittee Meeting (Clinical and Laboratory Standards Institute의 항생제 감수성 검사 소위원회 회의 소개)

  • Chang, Chulhun L.
    • Annals of Clinical Microbiology
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    • v.21 no.4
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    • pp.69-74
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    • 2018
  • Laboratory medicine is a specialized division that supports physicians in the care of patients by providing rapid and accurate in vitro diagnostic tests. Standardization of every component of a specific test is essential for producing accurate results. The Clinical and Laboratory Standards Institute (CLSI) was founded to develop a formal consensus process for standardization in 1968, and has been publishing standards and guidelines covering all aspects of clinical, research, and other laboratory work. CLSI guidelines are widely used around the world for standardization. The CLSI antimicrobial susceptibility testing subcommittee (AST SC) consists of 6 standing and many ad hoc working groups. Members of the AST SC review submitted proposals and suggestions, decide on approving these submissions in face-to-face meetings held twice a year, and revise CLSI documents accordingly. As these face-to-face meetings are open to anyone who registers to attend, I strongly encourage the members of our Society to attend and actively participate in document development.

Prevalence of pediculosis and scabies in preschool nursery children of Afyon, Turkey

  • CIFTCI Ihsan Hakki;KARACA Semsettin;DOGRU Omer;CETINKAYA Zafer;KULAC Mustafa
    • Parasites, Hosts and Diseases
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    • v.44 no.1 s.137
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    • pp.95-98
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    • 2006
  • Scabies and pediculosis are ubiquitous, contagious, and debilitating parasitic dermatoses. The tendency of high prevalence of pediculosis and scabies among school and preschool age children has prompted us to conduct a head louse and scabies prevalence survey among preschool nursery children in our district. A school-based, crosssectional study was performed, with 1,134 children chosen for evaluation. All cases were evaluated by physical examination and a detailed, structured questionnaire. The infestation was found in 14 $(1.2\%)$ of 1,134 children; 9 $(0.8\%)$ with pediculosis capitis and 5 $(0.4\%)$ with scabies. We found that infestations were more frequent in children with mothers whose education levels were low. This indicates the necessity of an improvement in the economic and sociocultural status of the community and the promotion of hygiene concepts and practices in order to improve health of preschool age children.

Synthesis of New Triazolyl-N,N-Dialkyldithiocarbamates as Antifungal Agents

  • Ozkirimli Sumru;Apak T. Idil;Kiraz Muammer;Yegenoglu Yildiz
    • Archives of Pharmacal Research
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    • v.28 no.11
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    • pp.1213-1218
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    • 2005
  • N,N-Dialkylditihiocarbamate derivatives have been well known as broad-range fungicides. In this study, the triazole derivatives of ten new N,N-disubstituted dithiocarbamates (3a-j) were synthesized and their structures were identified by spectral and elemental analysis. Results of the antifungal activity studies showed that some of the compounds tested were active against M. canis, M. gypseum, and T rubrum at the concentration of 12.5 $\mu$g/mL when c1otrimazol was used as a standard.

Antifungal Effect of Silver Nanoparticles on Dermatophytes

  • Kim, Keuk-Jun;Sung, Woo-Sang;Moon, Seok-Ki;Choi, Jong-Soo;Kim, Jong-Guk;Lee, Dong-Gun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1482-1484
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    • 2008
  • Spherical silver nanoparticles (nano-Ag) were synthesized and their antifungal effects on fungal pathogens of the skin were investigated. Nano-Ag showed potent activity against clinical isolates and ATCC strains of Trichophyton mentagrophytes and Candida species ($IC_{80}$, 1-7${\mu}g/ml$). The activity of nano-Ag was comparable to that of amphotericin B, but superior to that of fluconazole (amphotericin B $IC_{80}$, 1-5${\mu}g/ml$; fluconazole $IC_{80}$, 10-30${\mu}g/ml$). Additionally, we investigated their effects on the dimorphism of Candida albicans. The results showed nano-Ag exerted activity on the mycelia. Thus, the present study indicates nano-Ag may have considerable antifungal activity, deserving further investigation for clinical applications.

Antimicrobial Effect of Novel Pyrrolidinyl-thio Carbapenem, CW-270031 (신합성 카바페넴계 항생물질 CW-270031의 약효평가)

  • Kim, Jong-Myeung;Oh, Se-Woong;Ha, Jong-Ryul;Kim, Hong-Gi;Lee, Jin-Man;Lee, Sang-Han;Kim, Byoung-Oh;Kim, Jong-Guk
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.352-356
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    • 2006
  • CW-270031, an injectable carbapenem, is a novel synthesized pyrrolidinyl-thio carbapenems. It was evaluated for its in vitro antibacterial activities in comparison with those of imipenem and meropenem against standard strains and clinical isolated strains, CW-270031 was more active than imipenem against gram-negative (E. coli and Klebsiella oxytoca) clinical isolates, but it was slightly active than meropenem. Against Klebsiella aeruginosa CW-118 MIC were 0.048 $\mu$g/ml for CW-270031, 0.19 $\mu$g/ml for imipenem. Against clinical E. coli MIC range were 0.012$\sim$0.195 $\mu$g/ml for CW-270031, 0.097$\sim$0.39 $\mu$g/ml for imipenem. Against clinical Klebsiella oxytoca MIC$_{50}$ were 0.09 $\mu$g/ml for CW-270031, 0.39 $\mu$g/ml for imipenem. Against gram-positive standard strains and clinical CW-270031 was slightly more activity than meropenem, but CW-270033 was less active than imipenem against these tested isolates. The subcutaneous injection of CW-270031 in mice revealed that the half-life of CW-270031 in serum was about 13 min, long than that of meropenem (10.6 min). CW-270031. was stable to hydrolysis by dog renal dehydropeptidase I (DHP-l) enzyme, to an more stabilities shown by meropenem.

Antimicrobial Resistance and Molecular Epidemiologic Characteristics of Stenotrophomonas maltophilia Isolated from Clinical Specimens (병원 재료에서 분리한 Stenotrophomonas maltophilia의 항균제 내성 및 분자역학적 특성)

  • Seol, Sung-Yong;Jang, Kyoung-Soo;Jeong, Oung-Gi;Cho, Eung-Rae;Kim, Neung-Hee;Yu, Hak-Sun;Lee, Yoo-Chul;Cho, Dong-Taek
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.3
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    • pp.239-250
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    • 2000
  • Sixty-eight clinical isolates of Stenotrophomonas maltophilia from inpatients of 2 university hospitals in Taegu were epidemiologically analyzed by using the minimum inhibitory concentrations of 25 antimicrobial drugs, biochemical reaction, pulsed-field gel elctropgoresis (PFGE), and PCR with enterobacterial repetitive intergenic consensus sequences as primer (ERIC-PCR). 1. All the strains were susceptible to minocycline. More than 57% were susceptible to sulfisomidine (Su), ciprofloxacin (Ci), Ofloploxacin (Of), nalidixic acid (Na), and chloramphenicol (Cm), and $19{\sim}35%$ to ceftazidime (Cd), trimethoprim (Tp), Ticacillin-clavulanic acid, and cefoperazone-sulbactam. Most isolates were resistant to ${\beta}$-lactam antibiotics such as ampicillin (Ap), carbenicillin (Cb), cefotaxim (Ct), cefoxitin (Cx), and aminoglycosides including gentamicin (Gm), tobramycin (Tb), amikacin (Ak). 2. All the isolates were multiply resistant of 5 to 17 drugs and showed 40 different resistance pattern types. 3. All the strains showed very similar biochemical reactions except ${\beta}$-galactosidase and nitrate reduction test. Fourteen strains selected randomly were classified 10 different pattern type by PFGE and ERIC-PCR. These two methods showed identical result. Four strains isolated from wound in 1994 showed similar MIC pattern and identical API 20NE profile, PFGE, and ERIC-PCR pattern indicating episodes of cross-infection among patients. These results indicate that PFGE or ERIC-PCR profile has comparable discriminatory power for epidemiological typing of S. maltophilia.

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Intraspecific Variation of Environmental and Clinical Vibrio vulnificus Isolates as Demonstrated by Restriction Endonuclease Digestion Profiles

  • Kim, Ki-Yong;Yang, Ho-Chul;Tamplin, Mark-L.;Choi, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.78-83
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    • 1999
  • Thirty-six environmental isolates of Vibrio vulnificus obtained from seawater, sediments, and raw seafoods, and 18 clinical isolates from Vibrio septicemia patients were typed by restriction endonuclease digestion profiles (REDP) of genomic DNA with SfiI. The results revealed a high-level of variation in REDPs, indicating a vast genomic diversity among V. vulnificus strains. Genetic relatedness of the strains showed similarities ranging from 10% to 100%. Different REDPs for isolates from various raw seafoods were obtained, and clustering of strains according to type of seafoods was not observed. In contrast, clinical isolates of V. vulnificus showed higher similarity to one another, and could be subdivided into one separate group. The difference in REDPs of the V. vulnificus isolates from clinical origin and from raw seafoods substantiates the previous observation that only a single type of pathogenic strain was involved in each human infection, despite the numerous genetically polymorphic strains found from implicated oysters.

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