• Title/Summary/Keyword: polymerase chain reaction-sequence specific primers

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A Study of Genetic Polymonhisms of HLA-class I and II Genes Using Polymerase Chain Reaction (중합효소연쇄반응을 이용한 HLA-class I, II 유전자군의 유전적 다형성에 관한 연구)

  • Kyung-Ok Lee
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.11-25
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    • 1998
  • The HLA genes located in the short arm of chromosome 6 specify heterodimeric glycoproteins involved in the regulation of the immune response. Recently, in the elucidation of HLA polymorphism, serological and cellular typing methods have been replaced by DNA typing using polymerase chain reaction (PCR). The purpose of this study was to establish the HLA DNA typing methods and determine gene frequencies of HLA molecules in Koreans. PCR-SSP (sequence specific primers) and PCR-RFLP (restriction fragment length polymorphism) techniques were used for the analysis of HLA-A, -B, -C, DRBl genes and HLA-DQAl, DQBl, DPBl genes, respectively. The results of B-lymphoblastoid cells used for control experiment were consistent with the previous data identified in the 11th International Histocompatibility Workshop. Seventeen, 23, 16, 8, 16, 13 and 37 types of HLA-A, B, C, DQAl, DQBl, DPBl and DRBl alleles were found, respectively, in a total of unrelated 120 Korean individuals. The most frequent HLA alleles were $A^*$02 (27.0%), B$^*$40 (17.6%), Cw$^*$01 (19.2%), DQAl$^*$0301 (32.1%), DQBl$^*$0303 (12.9%), DPBl$^*$0501 (31.3%) and DRBl$^*$1501 (9.2%) among Koreans. This study shows that DNA typing method using PCR technique is a relatively simple, fast and practical tool for the determination of the HLA-class I and II genes. Moreover, the data of HLA gene frequencies could be useful for the Korean database before clinical applications, including organ and unrelated bone marrow transplantation, anthropological study, disease association and individual identification.

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Paprika Damping-off Outbreak Caused by Fusarium oxysporum Contaminated Seeds in Cheorwon Province in 2023 (2023 파프리카 종자의 Fusarium oxysporum 오염에 의한 철원지역 파프리카 모잘록병 대발생)

  • Miah Bae;Namsuk Kim;Sang Woo Kim;Sangyeon Ju;Byungyeon Kim;Soo Man Hwang;MeeKyoung Kim;Mi-Ri Park
    • Research in Plant Disease
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    • v.30 no.1
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    • pp.26-33
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    • 2024
  • In 2023, symptoms like damping-off disease were observed in 74 paprika growing in greenhouses in Cheorwon-gun, Gangwon-do, Korea. In this study, we tried to find the cause of the damping-off disease outbreak. We collected symptomatic seedlings and observed the typical crescent-shaped conidia of Fusarium oxysporum by microscope. To confirm the presence of F. oxysporum in the samples, polymerase chain reaction was performed using primers specific for F. oxysporum; the resulting sequence showed 99.11% identity with F. oxysporum. To confirm the pathogenicity of the F. oxysporum (CW) isolated from the samples, healthy paprika plants were inoculated with F. oxysporum CW and damping-off symptoms were observed 2 weeks later. To investigate whether the damping-off disease outbreak in Cheorwon-gun was caused by F. oxysporum-contaminated seeds, 100 paprika seeds were disinfected and placed in Murashige and Skoog medium. Typical pink F. oxysporum hyphae were found only in control non-disinfected seeds. An 18S rRNA-based and a TEF genebased phylogenetic analysis showed that the F. oxysporum CW isolate was not grouped with a F. oxysporum isolate reported from Cheorwon-gun in 2019. This study is the first report that an outbreak of damping-off disease in paprika in Cheorwon-gun, Gangwon-do, Korea, was caused by contamination of F. oxysporum seeds.

MHC Class II Allele Association in Korean Children With IgA Nephropathy and its Role as a Prognostic Factor (한국인 IgA 신병증 환아에서 MHC Class II유전자형과 예후와의 관계 분석)

  • Kim Pyung Kil;Yook Jinwon;Kim Ji Hong;Jang Yoon Soo;Shin Jeon-Soo;Choi In-Hong
    • Childhood Kidney Diseases
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    • v.4 no.1
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    • pp.33-39
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    • 2000
  • Purpose: Our study was designed to investigate the association of MHC Class II (DR, DQ) allele with IgA nephropathy and its significance as a prognostic factor for progression to ESRD Material and Methods: 69 children with IgA nephropathy with normal renal function(serum creatinine $\leq$ 1.5mg/dL) was classified as group A and 70 patients who received renal transplantation due to IgA nephropathy were selected as group B. The HLA-DQB1 and HLA-DRB1 alleles were studied by polymerase chain reaction using sequence specific primers. We have compared the difference in alleles between these two groups and with normal control and also examined any possible effect of the MHC class II genes on the histopathological severity and prognosis of IgAN. Results: Mean age was $8.8{\pm}2.9$ years in group A and $35.0{\pm}15.5$ years in group B. Male to female ratio was 2.8:1 in group A and 2.5:1 in group B. There was a significantly higher frequency of HLA-$DQB1^*03\;and\;DQB1^*05$ in Group B. The frequency of HLA-$DQB1^*0302\;and\;^*05031$ allele had increasing tendency in Group B(P<0.05). HLA-$DRB1^*03\;and\;^*05$ were more common in Group B(P<0.05). HLA-$DRB1^*04$ allele was the most common DR alleles in both group, but there was no statistical significance. There were no significant correlation with MHC class 13 genes on the hjstopathological severity in Group A. Conclusion: In conclusion, $HLA-DQB1^*0302\;and\;HLA-DQB1^*05031 $ allele seemed to be more common in transplanted patients compared to group with normal renal function suggesting that this allele is associated with poor prognosis in IgAN. However larger studies and follow up are required to confirm this due to uncharacterized heterogeneity in etiopathogenesis of IgA nephropathy and possibly one or more than one gene may exert influence in determining susceptibility to the diseases.

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