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Phylogenetic Analysis of env Gene V3-V5 Region of HIV-1 Subtype A Isolates from Korean (한국인 HIV 감염자에서 분리된 HIV-1 Subtype A의 env 유전자 V3-V5 부위의 계통적 분석)

  • Lee, Joo-Shil;Kim, Eun-Young;Kang, Chun;Nam, Jeong-Gu;Lee, Sung-Rae;Koo, Bon-Ki;Shin, Yung-Oh
    • The Journal of Korean Society of Virology
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    • v.29 no.2
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    • pp.119-127
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    • 1999
  • Phylogenetic analysis was conducted to monitor transmission of HIV and to investigate the genetic structure of primary isolates from 12 HIV-1 subtype A infected Koreans. The individuals infected with subtype A viruses had been diagnosed as HIV-1 seropositives during the period 1987 to 1995 and blood samples have been collected from 1991 to 1997. DNA of each individual was isolated from uncultured or cultured peripheral blood mononuclear cells. V3-V5 (0.7 kb) fragment of HIV-1 env gene was amplified by nested polymerase chain reaction and the PCR products were sequenced. The mean value of the divergence of nucleotide of HIV-1 env V3-V5 fragment was $17.0{\pm}4.06%$ ($8.6{\sim}25.8%$) within HIV-1 subtype A isolates from Koreans. This diversity was higher than those of African isolates ($13.7{\pm}2.66%$). In the phylogenetic tree, Korean subtype A isolates were not grouped together, but intermingled into African isolates. The results of this study suggested that HIV-1 subtype A variants be introduced from multiple sites of Africa into Korea and the big genetic diversity of Korea HIV-1 subtype A isolates may be further influenced by the range of geographic locations in which the infection occurred rather than the elapsed time between infection and collection of samples and the disease progression.

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Phylogenetic Characteristics of Echinococcus granulosus Sensu Lato in Uzbekistan

  • Kim, Hye-Jin;Yong, Tai-Soon;Shin, Myeong Heon;Lee, Kyu-Jae;Park, Gab-Man;Suvonkulov, Uktamjon;Kovalenko, Dmitriy;Yu, Hak Sun
    • Parasites, Hosts and Diseases
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    • v.58 no.2
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    • pp.205-210
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    • 2020
  • Echinococcosis occurs mainly in areas with heavy livestock farming, such as Central Asia, America, and Australia. Echinococcus granulosus sensu lato (s.l.) infection causes echinococcosis in intermediate hosts, such as sheep, cattle, goats, camels, and horses. Numerous cases of echinococcosis occur in Uzbekistan as stock farming is a primary industry. Epidemiological and genetic studies of E. granulosus s.l. are very important for mitigating its impact on public health and the economy; however, there are no such studies on E. granulosus s.l. in Uzbekistan. In the present study, to determine which genotypes exist and are transmitted, we isolated Echinococcus sp. from definitive hosts (one isolate each from jackal and dog) and intermediate hosts (52 isolates from humans and 6 isolates from sheep) in Uzbekistan and analyzed the isolates by sequencing 2 mitochondrial DNA components (cox1 and nad1). The results showed that all of isolates except one belonged to the E. granulosus sensu stricto (s.s.) G1 and G3 genotypes. Phylogenetic analysis based on cox1 sequences showed that 42 isolates from humans, 6 isolates from sheep, and one isolate from jackal were the G1 genotype, whereas the remaining 8 isolates from human and the one isolate from dog were the G3 genotype. These results suggest that the G1 and G3 genotypes of E. granulosus s.s. are predominant in Uzbekistan, and both wild animals and domestic animals are important for maintaining their life cycle. Only one isolate from human sample was confirmed to be E. eqiinus (G4 genotype), which is known to be for the first time.

Occurrence of Metalaxyl-Resistant Isolates of Pythium spp. Isolated from Turfgrasses of Golf Courses in Korea (우리나라 골프장 잔디에서 분리한 Metalaxyl 저항성 Pythium spp.의 발생)

  • Kim, Jin-Won;Park, Eun-Woo
    • The Korean Journal of Mycology
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    • v.30 no.2
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    • pp.157-165
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    • 2002
  • Of 125 isolates collected from 35 golf courses, sensitivity of 44 isolates of Pythium species to metalaxyl was determined on corn meal agar with various concentrations of metalaxyl (0.1, 1.0, 10.0, 50.0, 100.0, and $250.0{\mu}g\;a.\;i/ml$). The isolates were able to be categorized into the sensitive and resistant groups based on hyphal growth measured in terms of colony diameters on the medium with $1.0\;and\;10.0{\mu}g\;a.\;i./ml$. When compared with hyphal growth on the medium without metalaxyl, hyphal growth of the sensitive group which included 31 isolates was inhibited by $66{\sim}98%$ on the medium with $1.0{\mu}g\;a.\;i./ml$, whereas that of the resistant group which included 13 isolates grew well and the hyphal growth was inhibited only by $6{\sim}26%$. When $10.0{\mu}g\;a.\;i./ml$ of metalaxyl was included in the medium, hyphal growth of the sensitive and resistant groups was inhibited by $82{\sim}99%\;and\;27{\sim}47%$, respectively. Occurrence of metalaxyl-resistant isolates of Pythium spp. not only from turfgrasses on golf courses but also from other crops was observed for the first time in Korea. Metalaxyl-resistant isolates occurred most frequently in P. graminicola. Control effects of metalaxyl were determined by applying metalaxyl after and before inoculation of 4 and 3 isolates of sensitive and resistant isolates of P. graminicola, respectively, to creeping bentgrass in pots. The minimum concentration of metalaxyl to control metalaxyl-sensitive isolates was $6.25{\mu}g\;a.\;i./ml$, whereas the disease caused by the metalaxyl-resistant isolates could not be controlled with $12.50{\mu}g\;a.\;i./ml$ of metalaxyl. The disease was controlled more effectively by an application of metalaxyl prior to inoculation than after occurrence of the disease.

Identification of Mycobacteria by Comparative Sequence Apalysis and PCR-Restriction Fragment Length Polymorphism Analysis (염기서열과 PCR-Restriction Fragment Length Polymorphism 분석에 의한 Mycobacteria 동정)

  • Kook, Yoon-Hoh
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.6
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    • pp.561-571
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    • 1999
  • Diagnosis of mycobacterial infection is dependent upon the isolation and identification of causative agents. The procedures involved are time consuming and technically demanding. To improve the laborious identification process mycobacterial systematics supported by gene analysis is feasible, being particularly useful for slowly growing or uncultivable mycobacteria. To complement genetic analysis for the differentiation and identification of mycobacterial species, an alternative marker gene, rpoB encoding the ${\beta}$ subunit of RNA polymerase, was investigated. rpoB DNAs (342 bp) were amplified from 52 reference strains of mycobacteria including Mycobacterium tuberculosis H37Rv (ATCC 27294) and clinical isolates by the PCR. The nucleotide sequences were directly determined (306 bp) and aligned using the multiple alignment algorithm in the MegAlign package (DNASTAR) and MEGA program. A phylogenetic tree was constructed with a neighborhood joining method. Comparative sequence analysis of rpoB DNA provided the basis for species differentiation. By being grouped into species-specific clusters with low sequence divergence among strains belonging to same species, all the clinical isolates could be easily identified. Furthermore RFLP analysis enabled rapid identification of clinical isolates.

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Shot Hole of Peach and Japanese Plum Caused by Xanthomonas campestris pv. Pruni and Erwinia nigriflens in Korea (Xanthomonascampestris pv. pruni와 Erwinia nigrifluens에 의한 복숭아 및 자두의 세균성구멍병)

  • 최재을;이은정;박영섭
    • Research in Plant Disease
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    • v.6 no.1
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    • pp.10-14
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    • 2000
  • In 1998, bacterial shot hole of peach (Prunus persica) and Japanese plum(Prunus salicina) was found in Naju and Milyang. Five isolates of bacteria isolates from the diseased leaves and fruits of peach and Japanese plum were classified into genus Erwinia and Xanthomonas on diagnostic characteristics. Of five isolates, two were identified as X. campestris pv. pruni, three as E. nigrifluens. E.nigrifluens is the first description of bacteria which causes the disease on peach and Japanese plum in Korea. the symptoms caused by E. nigrifluens were hardly distinguished from those caused by X. campestris pv. pruni. In addition, it was observed that two pathogenic bacteria were isolated from most of naturally infected plants at the same time. from the reason mentioned above, we proposed to use a single common name \"bacterial shot hole of peach and Japanese plum\" for the both bacterial diseases, hereafter.

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Antagonistic and Plant Growth Promoting Activity of Bacillus species Isolated from Brackish Environment

  • Kamala-Kannan, Seralathan;Lee, Kui-Jae;Oh, Byung-Taek
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.05a
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    • pp.6-6
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    • 2010
  • Bacteria of the Bacillus sp. are well known to possess antagonistic activity against numerous plant pathogens. In the present study, 11 Bacillus sp. were isolated from the brackish environment and assayed for antagonistic activity under in vitro and in vivo conditions. Among the 11 isolates tested, 9 isolates effectively inhibited the growth of various plant pathogens, namely Phytophthora capsici, Phytophthora citropthora, Phytophthora citricola, Phytophthora sojae, Colletotricum coccodes, Colletotricum gloeosporioides, Colletotricum acutatum, Rhizoctonia solani, Fusarium solani, Fusarium graminearum, Pyricularia sp. and Monilina sp. The effective isolates were further screened for Phytophthora blight suppression in Capsicum annuum L. under green house conditions. The isolate SB10 exhibited the maximum (72.2%) reduction in disease severity. The antifungal compounds from the isolate were isolated and characterized. The isolated compounds exhibited high thermo stability ($100^{\circ}C$ for 30 min). Matrix-Assisted Laser Desorption Ionization-Time of Flight investigation of the antifungal compounds revealed three lipopeptide complexes, the surfactins, the iturins, and the fengycins.

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Evolution of Genetic Polymorphisms of Plasmodium falciparum Merozoite Surface Protein (PfMSP) in Thailand

  • Kuesap, Jiraporn;Chaijaroenkul, Wanna;Ketprathum, Kanchanok;Tattiyapong, Puntanat;Na-Bangchang, Kesara
    • Parasites, Hosts and Diseases
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    • v.52 no.1
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    • pp.105-109
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    • 2014
  • Plasmodium falciparum malaria is a major public health problem in Thailand due to the emergence of multidrug resistance. The understanding of genetic diversity of malaria parasites is essential for developing effective drugs and vaccines. The genetic diversity of the merozoite surface protein-1 (PfMSP-1) and merozoite surface protein-2 (PfMSP-2) genes was investigated in a total of 145 P. falciparum isolates collected from Mae Sot District, Tak Province, Thailand during 3 different periods (1997-1999, 2005-2007, and 2009-2010). Analysis of genetic polymorphisms was performed to track the evolution of genetic change of P. falciparum using PCR. Both individual genes and their combination patterns showed marked genetic diversity during the 3 study periods. The results strongly support that P. falciparum isolates in Thailand are markedly diverse and patterns changed with time. These 2 polymorphic genes could be used as molecular markers to detect multiple clone infections and differentiate recrudescence from reinfection in P. falciparum isolates in Thailand.

Rapid Detection of Methicillin Resistant Staphylococcus aureus Based on Surface Enhanced Raman Scattering

  • Han, Dae Jong;Kim, Hyuncheol
    • Korean Journal of Clinical Laboratory Science
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    • v.46 no.4
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    • pp.136-139
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    • 2014
  • Methicillin-resistant Staphylococcus aureus (MRSA) is one of the severe nosocomial infectious agents. The traditional diagnostic methods including biochemical test, antibiotic susceptibility test and PCR amplification are time consuming and require much work. The Surface enhanced Raman spectroscopy (SERS) biosensor is a rapid and powerful tool for analyzing the chemical composition within a single living cell. To identify the biochemical and genetic characterization of clinical MRSA, all isolates from patients were performed with VITEK2 gram positive (GP) bacterial identification and Antibiotic Susceptibility Testing (AST). Virulence genes of MRSA also were identified by DNA based PCR using specific primers. All isolates, which were placed on a gold coated nanochip, were analyzed by a confocal Raman microscopy system. All isolates were identified as S. aureus by biochemical tests. MRSA, which exhibited antibiotic resistance, demonstrated to be positive gene expression of both femA and mecA. Furthermore, Raman shift of S. aureus and MRSA (n=20) was perfectly distinguished by a confocal Raman microscopy system. This novel technique explained that a SERS based confocal Raman microscopy system can selectively isolate MRSA from non-MRSA. The study recommends the SERS technique as a rapid and sensitive method to detect antibiotic resistant S. aureus in a single cell level.

Detection of Extended-Spectrum β-Lactamase Producing Klebsiella pneumoniae by Multiplex Polymerase Chain Reaction (Multiplex Polymerase Chain Reaction을 이용한 Extended-Spectrum β-Lactamase 생성 Klebsiella pneumoniae 균주의 검출)

  • Yang, Byoung-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.173-178
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    • 2006
  • The production of extended-spectrum ${\beta}$-lactamases ($ESBL_S$) is the main mechanism of bacterial resistance to third-generation cephalosporins and monobactams, whose prevalence varies depending on the different geographical areas. In the last years it has increased notably to the point of being considered a health problem of great importance. The characterization of the ESBLs producing Klebsiella penumoniae strains present in clinical isolates is time-consuming. I describe here the development of a new system, which consists of a multiplex PCR. I found 51 K. pneumoniae strains to be presumptive strains ESBLs producers by clinical and laboratory standards institute (CLSI) guidelines. The double disc synergy test showed 47 positive K. pneumoniae, which were K. pneumoniae isolates. All ESBLs producing K. pneumoniae strains were resistant to antibiotic amikacin, gentamicin and ciprofloxacin. By multiplex PCR analysis, $bla_{TEM}$ gene in 17 strains 44 $bla_{SHV}$ genes and $bla_{CTX}$ genes in 33 strains were identified. In this study, the multiplex polymerase chain reaction (PCR) assay was a good method to detect and differentiate ESBLs producing K. penumoniae strains in clinical isolates.

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Screening of Trichoderma Isolates as a Biological Control Agent against Ceratocystis paradoxa Causing Pineapple Disease of Sugarcane

  • Rahman, M.A.;Begum, M.F.;Alam, M.F.
    • Mycobiology
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    • v.37 no.4
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    • pp.277-285
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    • 2009
  • In this study, dual culture, poison agar, and direct methods were used to assess the ability of Trichoderma virens IMI-392430, T. pseudokoningii IMI-392431, T. harzianum IMI-392432, T. harzianum IMI-392433, and T. harzianum IMI-392434 to control Ceratocystis paradoxa, which causes the pineapple disease of sugarcane. The highest percentage inhibition of radial growth (PIRG) values were observed with T. harzianum IMI-392432 using two dual culture methods, 63.80% in Method I and 80.82% in Method II. The minimum colony overgrowth time was observed with T. harzianum IMI-392432 and the maximum was observed with T. pseudokoningii IMI-392431. Different concentrations of different day-old metabolites of Trichoderma isolates were tested against mycelial growth of C. paradoxa. The highest PIRG (84.685%) exhibited at 80% concentration of 30-day-old metabolites of T. harzianum IMI-392432 using the modified bilayer poison agar method. In the direct assay method the maximum mycelial growth weight (PIGW) was observed at the same concentration and the same day-old metabolites of T. harzianum IMI-392432. This study showed that Trichoderma isolates have a good antagonistic effect on C. paradoxa mycelial growth and T. harzianum IMI-392432 has the most potential to control the pineapple disease pathogen.