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Journal of the Korean Society for Microbiology

  to  Present  ISSN: 0253-3162

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Detection and differentiation of Herpes Simplex Virus 1 and 2, and Varicella-Zoster virus in vesicle fluid, joint fluid and serum using PCR method.

Hae Kyung PARK ; So Youn WOO ; Hyun Jin KIM ; Chung Hwa LEE

Journal of the Korean Society for Microbiology.2000;35(2):191-201.

The viruses of Herpes Simplex Virus 1 (HSV-1), Herpes Simplex Virus 2 (HSV-2) and Varicella-Zoster virus (VZV) which belong to the alpha herpes subfamily are important human pathogens. When eruptions were not fully developed from these viral infections, clinical diagnosis was not always easy and required virological confirmation test. The above viruses were reactivated in individuals who were compromised in immune competence for one reason or another. Polymerase chain reaction (PCR) enables rapid and sensitive detection of HSV and VZV DNAs. Its sensitivity was largely influenced by choice of primers. Authors conducted a study to detect of those three viruses in human specimens including vesicle fluid and joint fluid and serum using PCR methods. Primers used for this study were the general primer pair GPHV-RU which was known to amplify within the genes enjoying the highest degree of homology between UL15 of HSV and UL42 of VZV. PCR with primers hybridized pair GPHV-RU amplifies a 396 bp with HSV-1 and HSV-2 standard stain DNA and 405 bp with VZV standard strain DNA. Restriction enzyme cleavage with HpaII and DdeI were used to detect and distinguish DNAs of HSV-1 and HSV-2 and VZV. The purpose of this study was a rapid and easy detection of VZV and HSV-1 or HSV-2 from various clinical specimens (vesicle fluid, serum and joint fluid) by PCR method. Used methods were: HSV PCR with primer 1, 2 and HpaII RE digestion; VZV nested PCR; HSV PCR with primer A, B and BssHII RE digestion. 1) In 33 cases (33/42, 78.6%) VZV was detected single or mixed infection from 42 clinical specimens which included vesicle fluid (5), serum from respiratory infected children (10), serum from immune suppressed adult cancer patients (7) and joint fluid from arthritis patients (20). 2) In 20 cases (20/42, 42.6%) HSV was detected singly or mixed infection and 19 of the cases were HSV-2 and 1 case was HSV-1. 3) In 19 cases (19/42, 45.2%) VZV was singly detected which included serum from respiratory infected children (6 cases), joint fluid from arthritis patients (9 cases), vesicle fluid (2 cases) and serum form immunosuppressed cancer patients (2 cases). 4) HSV was singly detected in 6 cases (6/42, 14.3%) which included joint fluid from arthritis patients (5 cases) and serum form respiratory infected children (1 cases). 5) 14 cases of VZV and HSV mixed infection (14/42, 33.3%) were detected. They included vesicle fluid (3 cases), serum form immunosuppressed cancer patients (4 cases), serum from respiratory infected children (2 cases) and joint fluid from arthritis patients (5 cases). 6) HSV-1 and HSV-2 detection and typing by HSV PCR with primer A, B and BssHII RE digestion method was more sensitive and the results were easier to detect than on other method.
Adult ; Arthritis ; Child ; Coinfection ; Diagnosis ; Digestion ; DNA ; Herpes Simplex* ; Herpesvirus 1, Human* ; Herpesvirus 2, Human ; Herpesvirus 3, Human* ; Humans ; Joints* ; Mental Competency ; Polymerase Chain Reaction* ; Simplexvirus*

Adult ; Arthritis ; Child ; Coinfection ; Diagnosis ; Digestion ; DNA ; Herpes Simplex* ; Herpesvirus 1, Human* ; Herpesvirus 2, Human ; Herpesvirus 3, Human* ; Humans ; Joints* ; Mental Competency ; Polymerase Chain Reaction* ; Simplexvirus*

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Detection of resistance mutation to lamivudine in HIV-1 infected patients.

Young Keol CHO ; Heung Sup SUNG ; Hee Jung LEE ; Yoo Kyum KIM ; Hyun Sook CHI ; Goon Jae CHO ; Moon Won KANG

Journal of the Korean Society for Microbiology.2000;35(2):181-190.

To investigate resistance to lamivudine (3TC), we examined the incidence of M184V in 20 HIV-1 patients treated with 3TC for 13.1 +/- 9 months. Fourteen of 20 patients had been exposed to zidovudine (ZDV) or didanosine (ddl) prior to 3TC therapy. Nested PCR targeting to reverse transcriptase (RT) and direct sequencing were performed for peripheral blood mononuclear cells sampled serially. There were resistance mutations to ZDV in at least 9 patients at baseline, although there was no resistance mutation to 3TC. We could detect M184V in 6 (30%) out of 20 patients. The incidence of M184V increased as the duration of therapy prolongs (13% in samples<12 months; 47% in samples gtoreq 12 months). The frequency of mutation M184V was higher in patients with previous mutation to ZDV than in patients with wild type. Resistance mutation was not detected in 7 patients. This study shows that resistance to 3TC tends to develop rapidly in patients with baseline mutations or two drugs combination therapy than in those treated simultaneously with triple drugs. This report is the first on resistance to 3TC in Korean AIDS patients.
Didanosine ; HIV-1* ; Humans ; Incidence ; Lamivudine* ; Polymerase Chain Reaction ; RNA-Directed DNA Polymerase ; Zidovudine

Didanosine ; HIV-1* ; Humans ; Incidence ; Lamivudine* ; Polymerase Chain Reaction ; RNA-Directed DNA Polymerase ; Zidovudine

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Pulsed field gel electrophoresis profile of erythromycin-clindamycin resistant Streptococcus pyogenes isolated in Korea.

Young Hee LEE ; Kyu Jam HWANG ; Kwang Jun LEE ; Kang Soo PARK ; Song Mee BAE ; Hwa Young SUNG ; Ki Sang KIM ; Chong Sam LEE

Journal of the Korean Society for Microbiology.2000;35(2):171-180.

Ninety two strains of Streptococcus pyogenes were isolated from patients with pharyngitis, scarlet fever, skin infection, and invasive streptococcal infections in Seoul, Korea from January to December, 1998. All isolates were epidemiologically characterized by T protein serotype, and serum opacity factor (OF) detection to phenotypes. To analyze the genetic relationship, fifty two isolates including 32 erythromycin-clindamycin (Em-Cm) resistant strains, 20 antimicrobial susceptible strains were attempted to the pulsed-field gel electrophoresis (PFGE). T protein serotype showed 16 kinds in distribution including T12 and T4. Among the total isolates, 40 strains (43.5%) belonged to the T12 serotype and twenty strains (21.7%) to T4 serotype. On the other hand, when infection aspect of S. pyogenes isolates were analysed by T serotype distribution, T12 type was predominant for pharyngitidis which contributed to 21 strains (53%) and for skin infection isolates which contributed to 11 strains (28%), respectively. In case of T4 type, it was the most predominant pharyngitidis isolates which contributed to 8 strains (40%). In T serotype distribution of Em-Cm resistant strains, 27 strains (84%) of the thirty two showed T12 serotype. In minimum inhibitory concentration (MIC) values of Em-Cm resistance isolates, thirty two isolates showed resistant to erythromycin 27 strains (84%), had high MIC of >128 mug/ml. And also to clindamycin, twenty two strains (69%) had high MIC of >128 mug/ml. When OF detection of Em-Cm resistance of S. pyogenes isolates were analyzed by T serotype distribution, T12 serotype isolates revealed that all of the isolates except one strain were OF negative. In PFGE profile analysis to Em-Cm resistance isolates, of the twenty seven, Em-Cm resistance of T12 serotype isolates, 26 strains showed identical PFGE profile and all of these isolates revealed that OF negative. Eighty four percent of Em-Cm resistance S. pyogenes isolates had identical phenotype and PFGE profile. These results strongly suggested that the Em-Cm resistant S. pyogenes isolates from Seoul area showed close genetic correlation and PFGE could be available tool for molecular epidemiology.
Clindamycin ; Electrophoresis, Gel, Pulsed-Field* ; Erythromycin ; Hand ; Humans ; Korea* ; Microbial Sensitivity Tests ; Molecular Epidemiology ; Pharyngitis ; Phenotype ; Scarlet Fever ; Seoul ; Skin ; Streptococcal Infections ; Streptococcus pyogenes* ; Streptococcus*

Clindamycin ; Electrophoresis, Gel, Pulsed-Field* ; Erythromycin ; Hand ; Humans ; Korea* ; Microbial Sensitivity Tests ; Molecular Epidemiology ; Pharyngitis ; Phenotype ; Scarlet Fever ; Seoul ; Skin ; Streptococcal Infections ; Streptococcus pyogenes* ; Streptococcus*

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Increased expression of adhesion molecules on human umbilical vein endothelial cells by Orientia tsutsugamushi infection.

Eun Bong LEE ; Seung Hoon HAN ; Sang Wook KIM ; Kyung Soo IHN ; Seung Yong SEONG ; Ik Sang KIM ; Myung Sik CHOI

Journal of the Korean Society for Microbiology.2000;35(2):159-169.

Scrub typhus is caused by Orientia tsutsugamushi characterized by fever, headache, lymphadenopathy and eschar formation. Infiltration of inflammatory cells around blood vessels and within the affected organs is known to be pathologic hallmark of the scrub typhus. Recently, expression of adhesion molecules on vascular endothelial cells was implicated as an important pathogenic mechanism in rickettsial disease. This study was performed to examine the expression of adhesion molecules and to investigate its role in the pathogenesis of O. tsutsugamushi infection. The expression of adhesion molecules on human umbilical vein endothelial cells (HUVEC) was measured by flow cytometry and indirect immunofluorescence. Expression of E-selectin, ICAM-1 and VCAM-1 was significantly increased 4 hours after the infection and persisted at least for 24 hours. Expression of those molecules was not induced by killed O. tsutsugamushi. Adhesion of polymorphonuclear cells and mononuclear cells to HUVEC was increased after the infection with O. tsutsugamushi. In conclusion, adhesion molecules are expressed on HUVEC during the infection of live O. tsutsugamushi and those molecules can contribute to the infiltration of inflammatory cells during the infection.
Blood Vessels ; E-Selectin ; Endothelial Cells ; Fever ; Flow Cytometry ; Fluorescent Antibody Technique, Indirect ; Headache ; Human Umbilical Vein Endothelial Cells* ; Humans* ; Intercellular Adhesion Molecule-1 ; Lymphatic Diseases ; Orientia tsutsugamushi* ; Scrub Typhus ; Vascular Cell Adhesion Molecule-1

Blood Vessels ; E-Selectin ; Endothelial Cells ; Fever ; Flow Cytometry ; Fluorescent Antibody Technique, Indirect ; Headache ; Human Umbilical Vein Endothelial Cells* ; Humans* ; Intercellular Adhesion Molecule-1 ; Lymphatic Diseases ; Orientia tsutsugamushi* ; Scrub Typhus ; Vascular Cell Adhesion Molecule-1

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Up-regulation of prothymosin alpha in THP-1 cells infected with Mycobacterium tuberculosis.

Ho Yeon SONG ; Kwang Sik JANG ; Hee Sun BYOUN ; Shin Je LEE ; Jin Koo KIM ; Yong Kyung CHOE ; Kwang Kjune KO

Journal of the Korean Society for Microbiology.2000;35(2):149-157.

Mycobacterium tuberculosis is capable of growing and survival within macrophage. The purpose of this study was to identify the genes regulated by infection of mycobacteria in human monocytic THP-1 cells. We used the differential display reverse transcriptase polymerase chain reaction (DD RT-PCR) and nothern blot analysis to confirm the differentially expressed genes from THP-1 cells infected with live Mycobacterium tuberculosis H37Rv, heat-kille Mycobacterium tuberculosis H37Rv and live Mycobacterium bovis BCG. Among many up or down-regulated clones, 27 clones were sequenced and compared with known genes on GenBank. Thirteen of over-expressed clones from THP-1 cells infected with live Mycobacterium tuberculosis H37Rv were identical to human prothymosin alpha, eight were novel clones and six clones showed homology with Human ferritin H chain, Escherichia coli bgl, Mouse RNA-dependent EIF-2 alpha kinase, E. coli htrL, Hyaluronan receptor and T cell receptor. Our result suggests that Mycobacterium tuberculosis might regulate prothymosin alpha gene transcription in monocytic THP-1 cell.
Animals ; Antigens, CD44 ; Clone Cells ; Databases, Nucleic Acid ; Escherichia coli ; Eukaryotic Initiation Factor-2 ; Ferritins ; Humans ; Macrophages ; Mice ; Mycobacterium bovis ; Mycobacterium tuberculosis* ; Mycobacterium* ; Phosphotransferases ; Receptors, Antigen, T-Cell ; Reverse Transcriptase Polymerase Chain Reaction ; Up-Regulation*

Animals ; Antigens, CD44 ; Clone Cells ; Databases, Nucleic Acid ; Escherichia coli ; Eukaryotic Initiation Factor-2 ; Ferritins ; Humans ; Macrophages ; Mice ; Mycobacterium bovis ; Mycobacterium tuberculosis* ; Mycobacterium* ; Phosphotransferases ; Receptors, Antigen, T-Cell ; Reverse Transcriptase Polymerase Chain Reaction ; Up-Regulation*

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Serovar Identification and Genetic Characterization of Leptospira Isolates by Arbitrarily Primed PCR and Ribotyping.

Woo Hyun CHANG ; Yoon Won KIM ; Hee Bok OH ; Min Kee CHO ; Sun Ho KEE ; Hyung Joon KIM

Journal of the Korean Society for Microbiology.1999;34(4):409-421.

Serovars of 22 leptospiral field isolates from rats trapped in Korea were identified by cross-agglutinin absorption test (CAAT). Genomic characteristics of 7 selected isolates and 6 antigenically closely related reference serovars of lai, yeonchon, birkini, gem, mwogolo, and canicola were differentiated by arbitrarily primed PCR (AP-PCR) and southern blot hybridization using 16S rRNA gene probe from Borrelia burgdorferi. Among the 22 isolates, 21 strains were identified as serovar lai by CAAT, while the serological reactivity of NR13 did not accord with that of serovar lai. Results of AP-PCR using primers RSP, KF and PB-1 were in general agreement with those obtained by serological identification, and all 7 isolates including NR13 showed the same profile with serovar lai or yeonchon. In the southern blot hybridization with 16S rRNA gene probe, the isolates were divided into two ribotype groups when HindIII and BamHI digests were employed: isolates NR4, NR13, and serovar lai showed the same profile, and isolates JR34, JR57, KR48, JR77, and JR82 were classified as the another ribotype group. Isolate NR13 and serovar yeonchon, which were isolated in Korea and showed serological differences with serovar lai, were indistinguishable from serovar lai in this DNA study using AP-PCR and ribotyping. These results demonstrate that Korean leptospiral isolates were closely related in DNA level, and ribotyping would be useful for subgrouping of field isolates.
Absorption ; Animals ; Blotting, Southern ; Borrelia burgdorferi ; DNA ; Genes, rRNA ; Korea ; Leptospira* ; Polymerase Chain Reaction* ; Rats ; Ribotyping*

Absorption ; Animals ; Blotting, Southern ; Borrelia burgdorferi ; DNA ; Genes, rRNA ; Korea ; Leptospira* ; Polymerase Chain Reaction* ; Rats ; Ribotyping*

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rpoB Gene Analysis of helicobacter pylori.

Kwang Ho RHEE ; Yoon Ho KOOK ; Myung Je CHO ; Seung Hyun LEE ; Bum Joon KIM ; Seo Jeong KIM ; Chang Young LIM ; Keun Hwa LEE ; Sun Ju YEO

Journal of the Korean Society for Microbiology.1999;34(4):401-408.

rpoB, which encodes the B subunit of RNA polymerase, is related to rifampin resistance of Mycobacterium tuberculosis and Escherichia coli. We determined the nucleotide sequences (346 bp) of rpoB gene from 25 Korean isolates of Helicobacter pylori. These nucleotide sequences were aligned and compared with H. pylori 26695 strain. No insertions or deletions were observed in all H. pylori strains. In the phylogenetic tree constructed by UPGMA method, 26 strains of H. pylori were separated into four clusters. Deduced amino acid sequences of amplified rpoB DNA comprised 115 amino acid residues. Twenty six H. pylori strains could be divided into 5 groups by the signature amino acid sequences. Two strains isolated from the same patient showed different nucleotide sequences. These results suggest that the sequences of rpoB are also highly divergent in H. pylori isolates and are useful for the epidemiologic study.
Amino Acid Sequence ; Base Sequence ; DNA ; DNA-Directed RNA Polymerases ; Escherichia coli ; Helicobacter pylori* ; Helicobacter* ; Humans ; Mycobacterium tuberculosis ; Rifampin

Amino Acid Sequence ; Base Sequence ; DNA ; DNA-Directed RNA Polymerases ; Escherichia coli ; Helicobacter pylori* ; Helicobacter* ; Humans ; Mycobacterium tuberculosis ; Rifampin

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Species Identification of the Genus Aeromonas Isolated from Freshwater Fish Culture-ponds and Seawater in Korea and Their Antibiotic Resistance Patterns.

Hun Ku LEE ; Dong Ryul KANG

Journal of the Korean Society for Microbiology.1999;34(4):393-400.

Some species of mesophilic bacteria in the genus Aeromonas were isolated from a snakehead (Channa argus) culture-pond in the Myung-Ji area, a freshwater culture-pond in Pukyung National University, and seawater near the sewage treatment plant in Kwang-An Beach in Pusan. They were identified by biological characteristics, and their susceptibility to ten antibiotics including ampicillin, cefoxitin, ceftazidime cephalothin, cefotiam, chloramphenicol, nalidixic acid, rifampin, cefoperazone-sulbactam, and imipenem, was also studied. Four species in the genus Aeromonas, A. veronii biovar sobria (27 strains), A. caviae (14 strains), A. trota (1 strain), and A. schubertii (1 strain) were identified. A. trota, an unrecorded species in Korea, is reported for the first time in this study. Three strains of A. veronii biovar sobria showed multiple resistance to three kinds of antibiotics and one strain to four drugs. All strains of A. caviae were resistant to more than three drugs and seven strains to four drugs. A. trota was resistant to cephalothin and rifampin, but A. schubertii showed multiple resistance to ampicillin, nalidixic acid, and rifampin.
Aeromonas* ; Ampicillin ; Animals ; Anti-Bacterial Agents ; Bacteria ; Busan ; Cefotiam ; Cefoxitin ; Ceftazidime ; Cephalothin ; Chloramphenicol ; Drug Resistance, Microbial* ; Fresh Water* ; Guinea Pigs ; Imipenem ; Korea* ; Nalidixic Acid ; Plants ; Population Characteristics ; Rifampin ; Seawater* ; Sewage

Aeromonas* ; Ampicillin ; Animals ; Anti-Bacterial Agents ; Bacteria ; Busan ; Cefotiam ; Cefoxitin ; Ceftazidime ; Cephalothin ; Chloramphenicol ; Drug Resistance, Microbial* ; Fresh Water* ; Guinea Pigs ; Imipenem ; Korea* ; Nalidixic Acid ; Plants ; Population Characteristics ; Rifampin ; Seawater* ; Sewage

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Molecular Characterization of papA Gene from Uropathogenic Escherichia coli.

Jong Bae KIM ; Hye Wone SONG

Journal of the Korean Society for Microbiology.1999;34(4):385-392.

The papA gene in uropathogenic Escherichia coli strains was characterized by PCR-restriction fragment length polymorphism (RFLP) and enterobacterial repetitive intergenic consensus (ERIC)- PCR. One hundred four E. coli strains from patients with urinary tract infections and 32 strains from healthy persons were examined. Thirty seven (27.2%) strains (33 from patients, 4 from healthy persons) in the 136 E. coli strains were positive in mannose resistant hemagglutination (MRHA) test. The adherence of MRHA positive strains to HEp-2 cells was greater than those of MRHA negative isolates (p<0.001). PapA-PCR were positive in 25% (26/104) of the strains from patients, and 3.1% (1/32) of the strains from healthy persons. Among 27 papA-positive isolates, subtypes were identified by RFLP as 8 (29.6%) F7z, 3 (11.1%) F9, 4 (14.8%) F12, and 4 (14.8%) F13. Six groups with novel RFLP patterns were detected, also. The subtypes of P-fimbriae was highly similar to each other by ERIC-PCR.
Consensus ; Hemagglutination ; Humans ; Mannose ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Urinary Tract Infections ; Uropathogenic Escherichia coli*

Consensus ; Hemagglutination ; Humans ; Mannose ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Urinary Tract Infections ; Uropathogenic Escherichia coli*

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Development of a Rapid Detection Method for Yersinia pestis by Polymerase Chain Reaction.

Ho Jung OH ; Hong Ki MIN ; Yeo Won SOHN ; Jeong Hoon CHUN ; Han Oh PARK

Journal of the Korean Society for Microbiology.1999;34(4):373-383.

A polymerase chain reaction (PCR) method for detection of the pathogenic Yersinia pestis from other Yersinia spp. was developed. Five Y. pestis strains, ninety-two other Yersinia species and twenty-four Enterobacteriaceae strains were collected in Korea and from other countries. Oligonucleotide primers were designed from pathogenic gene of antiphagocytic protein capsule gene (fra 1) and plasminogen activator gene (pla). The 428 bp DNA fragment was amplified from five Y. pestis which contained the fra I gene. No product was amplified from other Yersinia species and other strains of the Enterobacteriaceae. The 439 bp DNA fragment was amplified from three K pestis which contained the pla gene. No product was amplified from two Y. pestis, other Yersinia species and other strains of the Enterobacteriaceae. These showed that the designed primers were specific for detection of Y. pestis among other Yersinia species and Enterobacteriaceae strains. Amplification was successful whether the template was derived from purified DNA or from aliquots of boiled bacterial suspension. The detection limits were 100 pg of DNA and 100 colony forming units (CFU) for fra I and 100 pg DNA and 10 CFU for pla, respectively. Our results prove that the PCR method using specific primers for Y. pestis is a rapid and convenient procedure for routine clinical detection and identification of Y. pestis.
DNA ; DNA Primers ; Enterobacteriaceae ; Korea ; Limit of Detection ; Plasminogen Activators ; Polymerase Chain Reaction* ; Stem Cells ; Yersinia pestis* ; Yersinia*

DNA ; DNA Primers ; Enterobacteriaceae ; Korea ; Limit of Detection ; Plasminogen Activators ; Polymerase Chain Reaction* ; Stem Cells ; Yersinia pestis* ; Yersinia*

Country

Republic of Korea

Publisher

ElectronicLinks

Editor-in-chief

E-mail

Abbreviation

Journal of the Korean Society for Microbiology

Vernacular Journal Title

ISSN

0253-3162

EISSN

Year Approved

2007

Current Indexing Status

Currently Indexed

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Description

Current Title

Journal of Bacteriology and Virology

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