• Title/Summary/Keyword: M13 Virus

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First Report of Tomato Spotted Wilt Virus in Oxypetalum coeruleum in Korea (옥시페탈룸에서 발생한 토마토반점위조바이러스 국내 첫 보고)

  • Eseul, Baek;Peter, Palukaitis;Ju-Yeon, Yoon
    • Research in Plant Disease
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    • v.28 no.4
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    • pp.231-236
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    • 2022
  • Oxypetalum coeruleum, commonly known as Tweedia, is a perennial herbaceous plant of the Apocynaceae family native to southern Brazil and Uruguay. Tweedia plants are grown as one of the most popular ornamental flowers for floral arrangement in Korea. In May 2021, several tweedia plants in a single greenhouse in Gimje, Jeollabuk-do were found to show virus-like symptoms including necrotic rings, vein-clearing, chlorotic mottle, and mosaic on the leaves, and necrosis on the stems. Here, we have identified tomato spotted wilt virus (TSWV) in symptomatic tweedia leaves by applying high-throughput RNA sequencing. In the result, a single infection by TSWV was verified without mixed infections of different virus species. To confirm the presence of TSWV, a reverse transcription polymerase chain reaction was performed with a specific primer set to the N gene of TSWV. The complete genomic sequence of L, M, and S segments of TSWV 'Oxy' isolate were determined and deposited in GenBank under accession numbers LC671525, LC671638, and LC671639, respectively. In the phylogenetic tree analysis by maximum likelihood method, 'Oxy' isolate showed a high relationship with TSWV 'Gumi' isolate from Gerbera jamesonii in Gyeongsangbuk-do, Korea; for all three RNA segments. To our knowledge, this is the first report of TSWV infection of O. coeruleum in Korea.

Characterization of Immunogens of Infectious Hematopoietic Necrosis Virus Isolated in Korea (전염성 조혈기 괴사 바이러스(IHNV)의 항원 유도 단백질 특성)

  • Park, Myoung-Ae;Sohn, Sang-Gyu;Park, Jeong-Woo;Jeong, Young-Kee
    • Journal of fish pathology
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    • v.7 no.1
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    • pp.13-22
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    • 1994
  • To identify the immunogens of a PRT strain of Infectious Hematopoietci Necrosis Virus (IHNV) isolated from cultrued fish in Korea (Park et al, 1993). a panel of 4 monoclonal antibodies (MAbs) against IHNV-PRT strain and two polyclonal antisera from rainbow trout survived IHN disease were prepared. Proteins of purified IHNV-PRT strain were analysed on 10% SDS-PAGE and transferred onto NC paper and were incubated with the antibody solutions. With the polyclonal antibodies, four bands ($M_1$, $M_2$, G and 90Kd) were detected and the band density was in the order of $M_2$ > 90Kd > $M_1$ > G. However, with the MAbs, only two bands(G and 90Kd) were detected. The origin of 90Kd protein was not clear but maybe cell. All the results represented that among the five proteins of IHNV-PRT strain (Park et al., 1993), $M_2$, $M_1$ and G proteins were immunogens and $M_2$ protein was the strongest one.

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Plasma Real Time-Quantitative Polymerase Chain Reaction of Epstein-Barr Virus in Immunocompetent Patients with Hepatitis

  • Hong, Ji-Hye;Bae, Yon-Jung;Sohn, Joon-Hyung;Ye, Byung-Il;Chun, Jin-Kyong;Kim, Hwang-Min
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.15 no.1
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    • pp.38-43
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    • 2012
  • Purpose: Epstein-Barr virus (EBV) hepatitis is a usually asymptomatic and self-limiting disease in immunocompetent patients. However, the range of severity is wide, and the serological diagnosis is typically difficult until the convalescent phase. Thus, we examined the value of plasma EBV DNA real-time quantitative polymerase chain reaction (RT-qPCR) in EBV hepatitis for the timely diagnosis and the relationship between EBV viral load and clinical severity. Methods: Sixty samples were confirmed as having EBV infection by RT-qPCR with the EBV BALF5 gene sequence. We examined the clinical characteristics of EBV hepatitis by reviewing medical records. Results: The median total duration of fever was 8 days (range: 0-13 days). The mean peak value of aspartate aminotransferase (AST) was $241{\pm}214$ U/L, and the mean peak value of alanine aminotransferase (ALT) was $298{\pm}312$ U/L. There was no correlation between the serum levels of liver enzyme and plasma EBV DNA titer ($p$=0.1) or between median total duration of fever and EBV DNA titer ($p$=0.056). The median age of the EBV VCA IgM-negative group was lower compared with the EBV VCA IgM-positive group in EBV hepatitis (2 years vs. 6 years, $p$=0.0009). Conclusion: The severity of EBV hepatitis does not correlate with circulating EBV DNA load according to our data. Furthermore, we suggest that plasma EBV PCR may be valuable in young infants in whom the results of serology test for EBV infection commonly are negative.

Effect of Timing of IPTG Addition on Expression of Turnip Mosaic Virus Coat Protein Gene in Escherichia Coli (IPTG의 첨가 시간이 대장균(Escherichia coli)에서 순무 모자이크 바이러스(TuMV)의 외피단백질 발현에 미치는 영향)

  • Kim, Su-Joong;Park, Won-Mok;Ryu, Ki-Hyun;Lee, Sang-Seon;Lee, Se-Yong
    • Korean Journal Plant Pathology
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    • v.13 no.4
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    • pp.248-254
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    • 1997
  • Expression vector (pGEX-Tu) for the coat protein (CP) gene of turnip mosaic virus Ca strain (TuMV-Ca) was constructed by incorporation of TuMV CP gene into pGEX-KG vector which had ${\beta}$-galactosidase gene and IPTG (isopropylthio-${\beta}$-D-galactoside) induction site. The results of ELISA and western hybridization indicated that optimal condition of the expression were when IPTG and western hybridization indicated that optimal condition of the expression were when IPTG induction was carried out on YTA medium with ampicillin in 2 hours after the E. coli seed inoculation ($A_{595}$=0.1/ml). TuMV CP gene was expressed with GST (Glutathion S-Transferase) gene fusion system, and the size of fusion protein was estimated to be 59kDa, for TuMV CP was 33 kDa and GST was 26 kDa.

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Selection and identification of single-domain antibody against Peste des Petits Ruminants virus

  • Liu, Dan;Li, Lingxia;Cao, Xiaoan;Wu, Jinyan;Du, Guoyu;Shang, Youjun
    • Journal of Veterinary Science
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    • v.22 no.4
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    • pp.45.1-45.13
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    • 2021
  • Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Single-domain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 106 cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions: The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.

White Sport Syndrome Virus Disease of Shirmp and Diagnostic Methods

  • Zhan, Wen-Bin
    • Journal of Aquaculture
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    • v.15 no.1
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    • pp.7-13
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    • 2002
  • Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.

Analysis of HBeAg and HBV DNA Detection in Hepatitis B Patients Treated with Antiviral Therapy (항 바이러스 치료중인 B형 간염환자에서 HBeAg 및 HBV DNA 검출에 관한 분석)

  • Cheon, Jun Hong;Chae, Hong Ju;Park, Mi Sun;Lim, Soo Yeon;Yoo, Seon Hee;Lee, Sun Ho
    • The Korean Journal of Nuclear Medicine Technology
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    • v.23 no.1
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    • pp.35-39
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    • 2019
  • Purpose Hepatitis B virus (hepatitis B virus, HBV) infection is a worldwide major public health problem and it is known as a major cause of chronic hepatitis, liver cirrhosis and liver cancer. And serologic tests of hepatitis B virus is essential for diagnosing and treating these diseases. In addition, with the development of molecular diagnostics, the detection of HBV DNA in serum diagnoses HBV infection and is recognized as an important indicator for the antiviral agent treatment response assessment. We performed HBeAg assay using Immunoradiometric assay (IRMA) and Chemiluminescent Microparticle Immunoassay (CMIA) in hepatitis B patients treated with antiviral agents. The detection rate of HBV DNA in serum was measured and compared by RT-PCR (Real Time - Polymerase Chain Reaction) method Materials and Methods HBeAg serum examination and HBV DNA quantification test were conducted on 270 hepatitis B patients undergoing anti-virus treatment after diagnosis of hepatitis B virus infection. Two serologic tests (IRMA, CMIA) with different detection principles were applied for the HBeAg serum test. Serum HBV DNA was quantitatively measured by real-time polymerase chain reaction (RT-PCR) using the Abbott m2000 System. Results The detection rate of HBeAg was 24.1% (65/270) for IRMA and 82.2% (222/270) for CMIA. Detection rate of serum HBV DNA by real-time RT-PCR is 29.3% (79/270). The measured amount of serum HBV DNA concentration is $4.8{\times}10^7{\pm}1.9{\times}10^8IU/mL$($mean{\pm}SD$). The minimum value is 16IU/mL, the maximum value is $1.0{\times}10^9IU/mL$, and the reference value for quantitative detection limit is 15IU/mL. The detection rates and concentrations of HBV DNA by group according to the results of HBeAg serological (IRMA, CMIA)tests were as follows. 1) Group I (IRMA negative, CMIA positive, N = 169), HBV DNA detection rate of 17.7% (30/169), $6.8{\times}10^5{\pm}1.9{\times}10^6IU/mL$ 2) Group II (IRMA positive, CMIA positive, N = 53), HBV DNA detection rate 62.3% (33/53), $1.1{\times}10^8{\pm}2.8{\times}10^8IU/mL$ 3) Group III (IRMA negative, CMIA negative, N = 36), HBV DNA detection rate 36.1% (13/36), $3.0{\times}10^5{\pm}1.1{\times}10^6IU/mL$ 4) Group IV(IRMA positive, CMIA negative, N = 12), HBV DNA detection rate 25% (3/12), $1.3{\times}10^3{\pm}1.1{\times}10^3IU/mL$ Conclusion HBeAg detection rate according to the serological test showed a large difference. This difference is considered for a number of reasons such as characteristics of the Ab used for assay kit and epitope, HBV of genotype. Detection rate and the concentration of the group-specific HBV DNA classified serologic results confirmed the high detection rate and the concentration in Group II (IRMA-positive, CMIA positive, N = 53).

Analysis of the Factors Involved in the Occurrence of Rice Stripe virus in Chungcheongbukdo in 2008 and 2009 (2008-2009년 충북지역 벼 줄무늬잎마름병 발생 요인 분석)

  • Kang, Hyo-Jung;Ahn, Ki-Su;Han, Chong-U;Jeong, Kyeong-Heon;Park, See-Jung;Ji, Jae-Jun;Kim, Jeong-Soo
    • Research in Plant Disease
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    • v.16 no.2
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    • pp.109-114
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    • 2010
  • The occurrence factors of Rice stripe virus (RSV) in Chungcheongbukdo were analyzed by investigating the viruliferous insect rate (VIR) of overwintered small brown plant hopper (SBPH), the population density of SBPH, the infection rate of natural host plants, and the occurrence rate of RSV on rice paddy fields at the 3 areas of Cheongwon, Jincheon, and Boeun in 2008 and 2009. The average VIR of overwintered SBPH was 0.0% in 2008 and 1.1% in 2009. From SBPH collected on early June in 2009, VIR was higher as 1.4% at Jincheon and 4.2% at Boeun than those of overwintered SBPH, and this higher VIR might relate stronlgy with the adult population of SBPH immigrated from China. The populations of SBPH at Cheongwon, Jincheon and Boeun in 2008 were 3.8, 7.5 and 20.8 Head/$m^2$, respectively. However, those of Cheongwon and Jincheon increased up to about two folds as 8.4 and 13.1 in 2009. No RSV was detected on the natural host plants including barley. The factors involved in RSV occurrence were affected negatively by the low VIR of overwintered SBPH, the low population of overwintered SBPH, the low infection rate of RSV on the natural host plants, and the clean cultivation environment in Chungcheongbukdo.

Purification and Biological Activities of MT 1155 Inducing Morphological Change of Rous Sarcoma Virus-Transformed Normal Rat Kidney Cell (Rous Sarcoma Virus에 의해 형질전환된 NRK 세포의 형태변화를 유도하는 활성물질 MT 1154의 분리와 생물학적 활성)

  • 안종석;박문수;박찬선;윤병대;민태익;안순철;오원근;이현선;윤병대
    • Microbiology and Biotechnology Letters
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    • v.21 no.1
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    • pp.59-65
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    • 1993
  • We isolated Actinomycetes strain GMT 1155 and purified the active compound, MT 1155, on the morphological reversion of ts/NRK cell from the isolate. MT 1155 was identified as toyocamycin having antifungal and antitumor activities from physico-chemical properties and UV, IR, $^1H$-NMR, $^13C$-NMR and mass spectrum. MT 1155 showed the morphologically reversional activity on ts/NRK cell and the cytotoxicity on CTLL cell at the final concentrations of 1.7 JlM and 0.2 11M, respectively and its $IC_{50}$ value on protein kinase A enzyme was 2.3 $\mu$M. Also it had strong antifungal activity against several pathogenic fungi but not antibacterial activity. And it did not inhibit both protein kinase C activity and the bleb-formation of K562 cell induced by phorbol esters.

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Clinical and Serologic Study of Herpes Zoster in Children and Adolescents in Gwangju-Jeonnam Province (광주·전남 지역의 소아·청소년 대상포진에 대한 임상적 비교 및 혈청학적 의의에 대한 연구)

  • Na, Chan Ho;Kim, Choong Jae;Shin, Bong Seok;Choi, Hoon;Kim, Min Sung
    • Korean journal of dermatology
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    • v.56 no.9
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    • pp.531-538
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    • 2018
  • Background: Herpes zoster is a common dermatologic disorder. However, it rarely occurs in children and adolescents in Gwangju-Jeonnam province. Objective: This study aimed to analyze the epidemiological features and clinical characteristics of herpes zoster in children and adolescents. Methods: A retrospective survey was conducted on patients who visited the department of dermatology of Chosun University Hospital in Gwangju-Jeonnam province within the past 8 years. The medical records of 103 patients aged <18 years were reviewed. We analyzed for age, gender, accompanying symptoms, dermatomal distribution, underlying disease, treatment, and complications with serologic test. Results: The male-to-female ratio of the participants was 1.08:1, and their mean age was 13.0 years. Underlying diseases were observed in 3% (3/102) of the patients. The most common dermatomal distribution was thoracic dermatome (34%), followed by trigeminal (26%), cervical (20%), lumbar (15%), and non-skin (6%). The most common accompanying symptoms were headache (10%), fever (3%), and myalgia (3%). No difference was observed between patients who were varicella-zoster virus (VZV) IgM-positive and those who were VZV IgM-negative in terms of dermatome, visual analogue scale (VAS), severity, and body mass index (BMI). Conclusion: In children and adolescents with herpes zoster, the gender ratio and dermatomal distribution were similar to those previously reported, except for the low rate of underlying diseases. The incidence of herpes zoster in children was not significantly associated with immunosuppression and underlying diseases. Higher VZV IgM titer was not associated with dermatomal distribution, higher VAS score, or BMI. This study first compared the serological test results of children.