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Development of Nested PCR Primer Set for the Specific and Highly Sensitive Detection of Human Parvovirus B19

  • Cho, Kyu-Bong
    • Biomedical Science Letters
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    • v.24 no.4
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    • pp.390-397
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    • 2018
  • For the specific detection of human Parvovirus B19 (HuPaV-B19), we designed ten specific PCR primers from 3,800~4,500 nucleotides of HuPaV-B19 complete genome (NC_000883.2). Seventeen candidate PCR primer sets for specific detecting HuPaV-B19 were constructed. In specific reaction of HuPaV-B19, seventeen PCR primer sets showed specific band, however five PCR primer sets were selected basis of band intensity, amplicon size and location. In non-specific reaction with seven reference viruses, four PCR primer sets showed non-specific band, however one PCR primer set is not. Detection sensitivity of final selective PCR primer set was $100fg/{\mu}L$ for 112 minute, and PCR amplicon is 539 base pairs (bp). In addition, nested PCR primer set was developed, for detection HuPaV-B19 from a low concentration of contaminated samples. Selection of nested PCR primer set was basis of sensitivity and groundwater sample tests. Detection sensitivity of final selective PCR and nested PCR primer sets for the detection of HuPaV-B19 were $100fg/{\mu}L$ and $100ag/{\mu}L$ basis of HuPaV-B19 plasmid, it was able to rapid and highly sensitive detection of HuPaV-B19 than previous reports. We expect developed PCR primer set in this study will used for detection of HuPaV-B19 in various samples.

E1B-19k does not Localize in Mitochondria nor Dimerize Bax even with the Staurosporine (E1B-19k의 세포내 위치와 Bax와의 Dimerization에 관한 연구)

  • Yoon, Soo Han;Kim, Jin Young;Park, Seung Woo;Ahn, Young Hwan;Ahn, Young Min;Cho, Ki Hong;Cho, Kyung Gi
    • Journal of Korean Neurosurgical Society
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    • v.29 no.6
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    • pp.725-730
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    • 2000
  • Purpose : The subcellular localization of E1B-19k has been known cytosol or nuclear membrane by immunohistochemical staining and could dimerize with Bax to regulate cell death also known by the in-vitro immunoprecipitation. We planed to confirm this dimerization of E1B-19k with Bax in vivo in Cos-7 cells by using green fluorescent protein. Material and Method : We cloned E1B-19k and Bax into C3-EGFP. C3-EGFP-E1B-19k, C3-EGFP-Bax, and C3-EGFP-E1B-19k and pcDNA3-Bax were transfected into Cos-7 cells. We explored location of E1B-19k and Bax, and confirmed its dimerization with Bax in transfected living healthy Cos-7 cells by following green fluorescent protein of E1B-19k on the confocal microscope. Results : E1B-19k was located diffusely in cytoplasm and in nucleus but not in mitochondria. It prevented cell death from the apoptosis by staurosporine but its location was not changed. GFP-E1B-19k is not changed its intracellular location with Bax even with staurosporine. Conclusion : These results support that E1B-19k does not localize in mitochondria nor dimerize with Bax even with staurosporine. We could anticipate E1B-19k prevent cell death via the other dimerizing partner or pathways.

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Presence of Foxp3-expressing CD19(+)CD5(+) B Cells in Human Peripheral Blood Mononuclear Cells: Human CD19(+)CD5(+)Foxp3(+) Regulatory B Cell (Breg)

  • Noh, Joon-Yong;Choi, Wahn-Soo;Noh, Geun-Woong;Lee, Jae-Ho
    • IMMUNE NETWORK
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    • v.10 no.6
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    • pp.247-249
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    • 2010
  • Foxp3 is a transcript factor for regulatory T cell development. Interestingly, Foxp3-expressing cells were identified in B cells, especially in CD19(+)CD5(+) B cells, while those were not examined in CD19(+)CD5(-) B cells. Foxp3-expressing CD5(+) B cells in this study were identified in human PBMCs and were found to consist of $8.5{\pm}3.5%$ of CD19(+)CD5(+) B cells. CD19(+)CD5(+)Foxp3(+) B cells showed spontaneous apoptosis. Rare CD19(+)CD5(+) Foxp3(+) regulatory B cell (Breg) population was unveiled in human peripheral blood mononuclear cells and suggested as possible regulatory B cells (Breg) as regulatory T cells (Treg). The immunologic and the clinical relevant of Breg needs to be further investigated.

Aplastic Crisis Secondary to Parvovirus B19 Infection (Parvovirus B19 감염으로 발생된 Aplastic Crisis 3례)

  • Park, Yang Joon;Koh, Dae Kyun;Oh, Jin Hee
    • Clinical and Experimental Pediatrics
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    • v.46 no.11
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    • pp.1139-1142
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    • 2003
  • Human parvovirus(HPV) B19 infection causes erythema infectiosum in children, sometimes red cell aplastic crisis with hemolytic anemia and chronic bone marrow failure in immunocompromised hosts. HPV B19 is directly cytotoxic for erythroid progenitor cells and inhibits erythropoiesis. Infrequently, HPV B19 inhibits hematopoiesis of three cell lineages and causes transient pancytopenia in patients with hemolytic disorders. We report three patients with hereditary spherocytosis who developed transient aplastic crisis. A HPV B19 infection was confirmed by IgM anti-B19 parvovirus titers and characteristic findings of bone marrow examination as the causative agent associated with severe pancytopenia. Three patients recovered spontaneously after a short period of supportive care with red cell transfusions and intravenous immunoglobulin.

Phase Transformation and Reversible Shape Memory Effect of Ti-Ni-Cu Alloys (Ti-Ni-Cu 합금의 상변태 및 가역형상기억효과)

  • Hong, S.W.;Lee, O.Y.;Kim, D.K.
    • Journal of the Korean Society for Heat Treatment
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    • v.5 no.3
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    • pp.149-156
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    • 1992
  • Transformation behavior and reversible shape memory effct of Ti-Ni-Cu alloys with various Cu content has been investigated by means of electrical resistivity measurement, differential scanning calorimetry. X-ray diffraction and strain gage sensor. The transformation sequence in Ti-Ni-Cu alloys substituted by Cu for Ni up to 5at.% occurs to $B2{\leftrightarrow}B19^{\prime}$ and it proceeds in two stages by addition of 10 at.%Cu. i.e. $B2{\leftrightarrow}B19{\leftrightarrow}B19^{\prime}$. But the content of Cu increases up to 20at.%, it has been transformed in one stage ; $B2{\leftrightarrow}B19$. The shape change of Ti-40Ni-10Cu alloy which was constrain aged in circular form bended in $B2{\leftrightarrow}B19$ transformation but it spreaded out in $B19{\leftrightarrow}B19^{\prime}$ transformation. The amount of reversible shape change (${\Delta}{\varepsilon}$) of Ti-47Ni-3Cu alloy constrain aged at $400^{\circ}C$ after solution treatment has a maximum value of about $5.6{\times}10^{-3}$, but that of cold rolled and constrain aged specimens exhibits a little value independent of Cu concentrations.

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Clinical Characteristics of Human Parvovirus B19 Infection in Children

  • Jo, Kyo Jin;Lee, Yeoun Joo;Park, Kyung Mi;Yang, Eu Jeen;Yoo, Sukdong;Lim, Taek Jin;Park, Su Eun
    • Pediatric Infection and Vaccine
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    • v.27 no.2
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    • pp.111-116
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    • 2020
  • Purpose: Human parvovirus B19 infection is widespread and has a heterogeneous clinical spectrum, ranging from asymptomatic infection to potentially life-threatening complications. We investigated the various clinical features of human parvovirus B19 infection during an outbreak of the virus in our community. Methods: A retrospective chart review study was conducted at the Pusan National University Children's Hospital from December 2017 to April 2019. We investigated the clinical features of children with parvovirus B19 immunoglobulin M or parvovirus B19 DNA detected using polymerase chain reaction. Results: A total of 24 children were diagnosed with parvovirus B19 infection. Twelve (50%) had lace form rashes, and four (16.7%) had petechial rashes. Two (8.3%) were diagnosed with fever without a focus. Six (25%) developed aplastic crisis as a complication of infection, of whom three were previously diagnosed with hereditary spherocytosis and three with acute lymphoblastic leukemia. Conclusions: In addition to erythema infectiosum, the parvovirus B19 infection can present clinically with various types of rashes and fever without a focus. Furthermore, hematologic manifestations such as neutropenia and aplastic crisis can occur during infection.

Simultaneous Detection of Cytomegalovirus, Epstein-Barr Virus, Hepatitis B Virus, and Parvovirus by a Multiplex PCR (다중 중합효소 연쇄반응을 이용한 DNA 바이러스의 동시검출)

  • Sung, Hye-Ran;Joo, Jin-Young;Lee, Chong-Kil;Chung, Yeon-Bok;Song, Suk-Gil
    • Korean Journal of Microbiology
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    • v.43 no.1
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    • pp.1-6
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    • 2007
  • We describe a multiplex PCR method that can detect and differentiate simultaneously four different kinds of DNA viruses, Epstein-Barr virus (EBV), cytomegalovirus (CMV), hepatitis B virus (HBV) and parvovirus B19 (B19). Primers for the multiplex PCR reaction were designed to amplify specific regions of the EBV (pol), CMV (pol), HBV (pol) and B19 (ns) viral genomes and used to simultaneously detect individual viruses. In order to achieve optimal sensitivity and specificity for multiplex PCR, the thermo-cycling parameters, primer sequences, and concentration of each reaction components were optimized systematically. The sensitivity of the detection method ranged between 5 and 10 copies of viral genome with a mixture of multiple primer pairs. Furthermore, this highly sensitive test showed no cross-reactivity among the four viruses. Thus, the results obtained in this study provide evidence that the assay system is a good tool for supporting the diagnosis of viral infection and contamination.

Complete genome sequence of Streptococcus hyointestinalis B19, a strain producing bacteriocin, isolated from chicken feces

  • Lee, Ju-Eun;Heo, Sunhak;Kim, Geun-Bae
    • Journal of Animal Science and Technology
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    • v.62 no.3
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    • pp.420-422
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    • 2020
  • Streptococcus hyointestinalis B19 was isolated from chicken feces collected from local farm in Anseong, Korea. S. hyointestinalis B19 was shown to produce bacteriocin-like compounds exhibiting inhibitory activities against several pathogens including strains of Clostridium perfringens and Listeria monocytogenes. The whole genome of S. hyointestinalis B19 strain was sequenced using PacBio RS II platform. The genome comprised four contigs with a size of 2,217,061 bp. The DNA G + C content was found to be 42.95 mol%. Annotation results revealed 2,266 coding sequences (CDSs), 18 rRNAs, and 61 tRNA genes. Based on genome analysis, we found that the strain B19 possessed various genes associated with bacteriocin synthesis, modification, and transport.

Selection of Beneficial Microbial Agents for Control of Fungal Diseases in the Phyllosphere of Cucumber Plant (오이 지상부의 주요 곰팡이 병해의 생물적 방제용 유용미생물의 선발)

  • Lee, Sang-Yeob;Lee, Young-Kee;Park, Kyung-Seok;Kim, Yong-Ki
    • The Korean Journal of Pesticide Science
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    • v.14 no.4
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    • pp.326-331
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    • 2010
  • Bacillus subtilis B29, B. subtilis M10 and Streptomyces sp. CC19 obtained from phyllosphere of cucumber plants were selected for biological control of fungal air-borne diseases. For the downy mildew, diseased area of B. subtilis B29, B. subtilis M10 and Streptomyces sp. CC19 showed 0.5%, 20.2% and 42.0%, but that of control was 82.0% respectively, in cucumber seedling test. Incidence of powdery mildew by once application of B. subtilis B29, B. subtilis M10 and Streptomyces sp. CC19 was 2.8%, 3.6% and 12.3%, respectively, whereas that of control was 65.6%. On the gray mold, diseased area of B. subtilis B29, B. subtilis M10 and Streptomyces sp. CC19 was 8.0%, 30.8% and 5.2%, respectively, compared to 81.2% for the control. Therefore, B. subtilis B29 could be a prospective antagonist for biological control of powdery mildew, downy mildew and gray mold of cucumber plant.

Parvovirus B19 Infection in Two Korean Pediatric Kidney Transplant Patients (소아에서 신장 이식 후 발생한 Parvovirus Bl9 감염 2례)

  • Koo, So-Eun;Lee, Joo-Hoon;Hahn, Hye-Won;Han, Duck-Jong;Park, Young-Seo
    • Childhood Kidney Diseases
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    • v.9 no.2
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    • pp.275-281
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    • 2005
  • Parvovirus B19(PV B19) is a nonenveloped single-stranded DNA virus that causes a wide variety of diseases ranging from benign childhood infection such as slapped-cheek rash(fifth disease) to life threatening diseases such as hydrous fetalis in fetuses or aplastic anemic crises in patients with hemolytic anemia. In immunocompromised hosts including organ transplant recipients, this infection can cause chronic anemia. Recently, the reports of cases of PV B19 infection have been increasing in transplant recipients and most reported cases of PV B19 infection-associated anemia in renal transplant recipients were successfully treated with intravenous immunoglobulin infusion. We experienced two cases of aplastic anemia caused by PV B19 infection in pediatric renal transplant recipients. The patients were an 8 year-old boy and 12-year-old boy who received allograft kidneys from their mothers. Anemia developed 2 weeks after transplantation and their serum was positive for PV B19 PCR. They were treated with 400 mg/kg of intravenous immunoglobulin(IVIG) for 5 consecutive days. In one of the case, anemia was corrected promptly after the first 5-day course of IVIG therapy but in the other, anemia persisted but responded to the second course of IVIG therapy. One year later, the patients have normal hematocrit levels and stable renal function These are the first cases of PV B19 infection treated successfully with IVIG in pediatric renal transplant recipients in Korea. (J Korean Soc Pediatr Nephrol 2005;9:275-281)

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