• Title/Summary/Keyword: virus assay

Search Result 687, Processing Time 0.024 seconds

The concentration of serum amyloid A and haptoglobin of pigs infected with Aujeszky′s disease virus (오제스키병에 감염된 돼지의 serum amyloid A와 haptoglobin의 농도 변화)

  • 오윤택;조정곤
    • Korean Journal of Veterinary Service
    • /
    • v.25 no.1
    • /
    • pp.45-52
    • /
    • 2002
  • The acute phase serum protein response is a well-known general indicator of inflammation, trauma or other pathological conditions and its relevance for the monitoring of the health status of domestic animals is being increasingly realized. The changes in serum protein composition which occur after tissue damage represent a part of the systemic response of the injured animals which is mediated by pro-inflammatory cytokines such as TNF-$\alpha$, IL-6 and IL-1. These responses play a vital role in containing the tissue damage and enhancing the processes of repair and resolution. From a clinical perspective, the assay of acute phase proteins can provide a method for detecting inflammation. In animals, the most sensitive acute phase proteins are haptoglogin, serum amyloid A and at-acid glycoprotein in response to inflammatory condition. The aim of this study was to assess the diagnostic value of the concentrations of serum amyloid A(SAA) and haptoglobin(HP) in serum of pigs infected with Aujeszky's disease virus(ADV). Fifty pigs infected with ADV and 5 normal pigs were used in this experiment. The mean serum concentration of Shh of pigs infected with ADV was 96.8 $\pm$ 7.1 $\mu\textrm{g}$/㎖(range, 36.0∼187.5 $\mu\textrm{g}$/㎖) and that of normal pigs was 42.9$\pm$3.3 $\mu\textrm{g}$/㎖(range, 17.3∼127.8 $\mu\textrm{g}$/㎖). The mean serum concentration of HP of pigs infected with ADV was 1,164.4 $\pm$ 96.9 $\mu\textrm{g}$/㎖ (range, 790.2∼l,769.2 $\mu\textrm{g}$/㎖) and that of normal pigs was 675.4 $\pm$ 56.3 $\mu\textrm{g}$/㎖ (range, 650.0-690.4 $\mu\textrm{g}$/㎖). The mean concentrations of SAA and HP in serum of pigs infected with ADV compared with those of normal pigs showed approximately a two-fold. It was concluded that the concentrations of Shh and HP in serum may proved to be diagnostic marker of Aujeszky's disease.

Ribavirin Does Not Impair the Suppressive Activity of $Foxp3^+$ $ CD4^+$ $CD25^+$ Regulatory T Cells

  • Lee, Jeewon;Choi, Yoon Seok;Shin, Eui-Cheol
    • IMMUNE NETWORK
    • /
    • v.13 no.1
    • /
    • pp.25-29
    • /
    • 2013
  • Ribavirin is an antiviral drug used in combination with pegylated interferon-${\alpha}$ (IFN-${\alpha}$) for the treatment of hepatitis C virus (HCV) infection. Recently, ribavirin was reported to inhibit the suppressive activity of regulatory T (Treg) cells. In the present study, we re-evaluated the effect of ribavirin on $CD4^+$ $CD4^+$ $CD25^+$ Treg cells from normal donors. First, we examined the expression of CTLA-4 and CD39, which are known to play a role in the suppressive function of Treg cells. We found that ribavirin treatment did not modulate the expression of CTLA-4 and CD39 in Treg cells. We also studied the effect of ribavirin on Treg cells in the presence of IFN-${\alpha}$; however, the expression of CTLA-4 and CD39 in Treg cells was not changed by ribavirin in the presence of IFN-${\alpha}$. Next, we directly evaluated the effect of ribavirin on the suppressive activity of Treg cells in the standard Treg suppression assay, by co-culturing CFSE-labeled non-Treg $CD4^+$ T cells with purified Treg cells. We found that ribavirin did not attenuate the suppressive activity of Treg cells. Taken together, while ribavirin reversed Treg cell-mediated suppression of effector T cells in the previous study, we herein demonstrate that ribavirin does not impair the suppressive activity of Treg cells.

Azasugar-Containing Phosphorothioate Oligonucleotide (AZPSON) DBM-2198 Inhibits Human Immunodeficiency Virus Type 1 (HIV-1) Replication by Blocking HIV-1 gp120 without Affecting the V3 Region

  • Lee, Jinjoo;Byeon, Se Eun;Jung, Ju Yeol;Kang, Myeong-Ho;Park, Yu-Jin;Jung, Kyeong-Eun;Bae, Yong-Soo
    • Molecules and Cells
    • /
    • v.38 no.2
    • /
    • pp.122-129
    • /
    • 2015
  • DBM-2198, a six-membered azasugar nucleotide (6-AZN)-containing phosphorothioate (P = S) oligonucleotide (AZPSON), was described in our previous publication [Lee et al. (2005)] with regard to its antiviral activity against a broad spectrum of HIV-1 variants. This report describes the mechanisms underlying the anti-HIV-1 properties of DBM-2198. The LTR-mediated reporter assay indicated that the anti-HIV-1 activity of DBM-2198 is attributed to an extracellular mode of action rather than intracellular sequence-specific antisense activity. Nevertheless, the antiviral properties of DBM-2198 and other AZPSONs were highly restricted to HIV-1. Unlike other P = S oligonucleotides, DBM-2198 caused no host cell activation upon administration to cultures. HIV-1 that was pre-incubated with DBM-2198 did not show any infectivity towards host cells whereas host cells pre-incubated with DBM-2198 remained susceptible to HIV-1 infection, suggesting that DBM-2198 acts on the virus particle rather than cell surface molecules in the inhibition of HIV-1 infection. Competition assays for binding to HIV-1 envelope protein with anti-gp120 and anti-V3 antibodies revealed that DBM-2198 acts on the viral attachment site of HIV-1 gp120, but not on the V3 region. This report provides a better understanding of the antiviral mechanism of DBM-2198 and may contribute to the development of a potential therapeutic drug against a broad spectrum of HIV-1 variants.

Detection of foot-and-mouth disease virus (FMDV) and avian influenza virus (AIV) from animal carcass disposal sites using real-time RT-PCR

  • Miguel, Michelle;Kim, Seon-Ho;Lee, Sang-Suk;Cho, Yong-Il
    • Korean Journal of Veterinary Service
    • /
    • v.43 no.2
    • /
    • pp.107-112
    • /
    • 2020
  • Foot-and-mouth disease (FMD) and avian influenza (AI) are highly pathogenic viral disease which affects the livestock industry worldwide. Outbreak of these viruses causes great impact in the livestock industry; thus, disease infected animals were immediately disposed. Burial is the commonly used disposal method for deceased animals. However, there is potential for secondary environmental contamination, as well as the risk that infectious agents persisting in the environment due to the limited environmental controls in livestock burial sites during the decomposition of the carcasses. Therefore, this study aimed to investigate the detection of FMD and AI viruses from animal carcass disposal sites using real-time reverse transcription PCR. Soil samples of more than three years post-burial from livestock carcass disposal sites were collected and processed RNA isolation using a commercial extraction kit. The isolated RNA of the samples was used for the detection of FMDV and AIV using qRT-PCR. Based on the qPCR assay result, no viral particle was detected in the soil samples collected from the animal disposal sites. This indicates that 3 years of burial and their carcass disposal method is efficient for the control or at least reduction of spread infections in the surrounding environment.

Diagnosis and gI antibody dynamics of pseudorabies virus in an intensive pig farm in Hei Longjiang Province

  • Wang, Jintao;Han, Huansheng;Liu, Wanning;Li, Shinian;Guo, Donghua
    • Journal of Veterinary Science
    • /
    • v.22 no.2
    • /
    • pp.23.1-23.10
    • /
    • 2021
  • Background: Pseudorabies (PR), caused by the pseudorabies virus (PRV), is an endemic disease in some regions of China. Although there are many reports on epidemiological investigations into pseudorabies, information on PRV gI antibody dynamics in one pig farm is sparse. Objectives: To diagnose PR and analyze the course of PR eradication in one pig farm. Methods: Ten brains and 1,513 serum samples from different groups of pigs in a pig farm were collected to detect PRV gE gene and PRV gI antibody presence using real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Results: The July 2015 results indicated that almost all brain samples were PRV gE gene positive, but PRV gI antibody results in the serum samples of the same piglets were all negative. In the boar herd, from October 2015 to July 2018 three positive individuals were culled in October 2015, and the negative status of the remaining boars was maintained in the following tests. In the sow herd, the PRV gI antibody positive rate was always more than 70% from October 2015 to October 2017; however, it decreased to 27% in January 2018 but increased to 40% and 52% in April and July 2018, respectively. The PRV gI antibody positive rate in 100-day pigs markedly decreased in October 2016 and was maintained at less than 30% in the following tests. For 150-day pigs, the PRV gI antibody positive rate decreased notably to 10% in April 2017 and maintained a negative status from July 2017. The positive trend of PRV gI antibody with an increase in pig age remarkably decreased in three tests in 2018. Conclusions: The results indicate that serological testing is not sensitive in the early stage of a PRV infection and that gilt introduction is a risk factor for a PRV-negative pig farm. The data on PRV gI antibody dynamics can provide reference information for pig farms wanting to eradicate PR.

Identification and Expression Analysis of Genes Induced in Response to Tomato chlorosis virus Infection in Tomato

  • Sahin-Cevik, Mehtap;Sivri, Emine Dogus;Cevik, Bayram
    • The Plant Pathology Journal
    • /
    • v.35 no.3
    • /
    • pp.257-273
    • /
    • 2019
  • Tomato (Solanum lycopersicum) is one of the most widely grown and economically important vegetable crops in the world. Tomato chlorosis virus (ToCV) is one of the recently emerged viruses of tomato distributed worldwide. ToCV-tomato interaction was investigated at the molecular level for determining changes in the expression of tomato genes in response to ToCV infection in this study. A cDNA library enriched with genes induced in response to ToCV infection were constructed and 240 cDNAs were sequenced from this library. The macroarray analysis of 108 cDNAs revealed that the expression of 92 non-redundant tomato genes was induced by 1.5-fold or greater in response to ToCV infection. The majority of ToCV-induced genes identified in this study were associated with a variety of cellular functions including transcription, defense and defense signaling, metabolism, energy, transport facilitation, protein synthesis and fate and cellular biogenesis. Twenty ToCV-induced genes from different functional groups were selected and induction of 19 of these genes in response to ToCV infection was validated by RT-qPCR assay. Finally, the expression of 6 selected genes was analyzed in different stages of ToCV infection from 0 to 45 dpi. While the expression of three of these genes was only induced by ToCV infection, others were induced both by ToCV infection and wounding. The result showed that ToCV induced the basic defense response and activated the defense signaling in tomato plants at different stages of the infection. Functions of these defense related genes and their potential roles in disease development and resistance to ToCV are also discussed.

Status of Research and Development of Foot and Mouth Disease Diagnosis (Review) (구제역 진단법 연구개발 현황 (총설))

  • Kwak, Kyeongrok;Choi, So-Young;Kim, Eunyoung;Hwang, Choon Hong;Lee, Sung-Jin
    • ANNALS OF ANIMAL RESOURCE SCIENCES
    • /
    • v.28 no.2
    • /
    • pp.78-96
    • /
    • 2017
  • Foot-and-mouth disease (FMD) is a infection that can easily spread when it occurs and causes serious economic damage because of the existence of multiple serotypes of the virus and extreme contagiousness. The most effective method in preventing the transmission of FMD virus (FMDV) is the culling of livestock and additional vaccination in the other areas depending on the spreading rate and situation. Diagnostic methods are utilized not only for the definite diagnosis of FMD but also for identification of serotype, and confirmation of antibody production after vaccination. Although many methods have been developed to diagnose, they are not still enough to detect accurately the disease in a short time. Therefore, it has been needed new diagnostic methods improved from existing methods. Previous methods were based on the enzyme-linked immunosorbent assay (ELISA) as a serological diagnostic method, or polymerase chain reaction (PCR), which is a molecular genetic method. The recent technology has been performing about the combination of both methods and how to make it faster, less costly, more sensitive and accurate way.

Development of reverse transcription loop-mediated isothermal amplification assays for point-of-care testing of avian influenza virus subtype H5 and H9

  • Zhang, Songzi;Shin, Juyoun;Shin, Sun;Chung, Yeun-Jun
    • Genomics & Informatics
    • /
    • v.18 no.4
    • /
    • pp.40.1-40.8
    • /
    • 2020
  • Avian influenza (AIV) outbreaks can induce fatal human pulmonary infections in addition to economic losses to the poultry industry. In this study, we aimed to develop a rapid and sensitive point-of-care AIV test using loop-mediated isothermal amplification (LAMP) technology. We designed three sets of reverse transcription LAMP (RT-LAMP) primers targeting the matrix (M) and hemagglutinin (HA) genes of the H5 and H9 subtypes. RT-LAMP targeting the universal M gene was designed to screen for the presence of AIV and RT-LAMP assays targeting H5-HA and H9-HA were designed to discriminate between the H5 and H9 subtypes. All three RT-LAMP assays showed specific amplification results without nonspecific reactions. In terms of sensitivity, the detection limits of our RT-LAMP assays were 100 to 1,000 RNA copies per reaction, which were 10 times more sensitive than the detection limits of the reference reverse-transcription polymerase chain reaction (RT-PCR) (1,000 to 10,000 RNA copies per reaction). The reaction time of our RT-LAMP assays was less than 30 min, which was approximately four times quicker than that of conventional RT-PCR. Altogether, these assays successfully detected the existence of AIV and discriminated between the H5 or H9 subtypes with higher sensitivity and less time than the conventional RT-PCR assay.

Development of indirect ELISA for the detection of canine adenovirus type 2 antibodies in dog sera

  • Yang, Dong-Kun;Kim, Ha-Hyun;Lee, Siu;Oh, Dongryul;Yoo, Jae Young;Hyun, Bang-Hun
    • Journal of Veterinary Science
    • /
    • v.21 no.4
    • /
    • pp.63.1-63.9
    • /
    • 2020
  • Background: Canine adenovirus type 2 (CAV-2) induces infectious laryngotracheitis in members of the family Canidae, including dogs. To date, no ELISA kits specific for CAV-2 antibody have been commercialized for dogs in Korea. Objectives: We aimed to develop new indirect enzyme-linked immunosorbent assay (I-ELISA) to perform rapid, accurate serological surveys of CAV-2 in dog serum samples. Methods: In total, 165 serum samples were collected from dogs residing in Chungbuk and Gyeongbuk provinces between 2016 and 2018. The Korean CAV-2, named the APQA1701-40P strain, was propagated in Madin-Darby canine kidney cells and purified in an anion-exchange chromatography column for use as an antigen for I-ELISA. The virus-neutralizing antibody titers of CAV-2 in the dog sera were measured by virus neutralization (VN) test. Results: We compared the results obtained between the VN and new I-ELISA tests. The sensitivity, specificity, and accuracy of new I-ELISA were 98.6%, 86.4% and 97.0% compared with VN test, respectively. New I-ELISA was significantly correlated with VN (r = 0.91). Conclusions: These results indicate that new I-ELISA is useful for sero-surveillance of CAV-2 in dog serum.

Biological and Molecular Characterization of a Korean Isolate of Orthotospovirus chrysanthinecrocaulis (Formerly Chrysanthemum Stem Necrosis Virus) Isolated from Chrysanthemum morifolium

  • Seong Hyeon Yoon;Su Bin Lee;Eseul Baek;Ho-Jong Ju;Ju-Yeon Yoon
    • Research in Plant Disease
    • /
    • v.29 no.3
    • /
    • pp.286-294
    • /
    • 2023
  • Biological and molecular characterization of a Korean isolate of Orthotospovirus chrysanthinecrocaulis (formerly known as chrysanthemum stem necrosis virus, CSNV) isolated from Chrysanthemum morifolium was determined using host range and sequence analysis in this study. Twenty-three species of indicator plants inoculated mechanically CSNV-Kr was investigated for determination of host range. CSNV-Kr induced various local and systemic symptoms in the inoculated plant species. CSNV-Kr could not infect three plant species and induced symptomless in systemic leaves in Nicotiana tabacum cultivars, though the plant samples reacted positively with the antiserum to CSNV by double-antibody sandwich-enzyme-linked immunosorbent assay. The complete genome sequence of CSNV-Kr was determined. The L RNA of CSNV-Kr consists of 8,959 nucleotides (nt) and encodes a putative RNA-dependent RNA polymerase. The M RNA of CSNV-Kr consists of 4,835 nt and encodes the movement protein (NSm) and the glycoprotein precursor (Gn/Gc protein). The S RNA of CNSV-Kr consists of 2,836 nt and encodes NSs protein and N protein. The Gn/Gc and N sequence of CSNV-Kr were compared with those of previously published CSNV isolates originating from different countries at nucleotide and amino acid levels. The Gn/GC sequence of CSNV-Kr shared 98.8-99.5% identity with CSNV isolated from other countries and the N sequence of CSNV-Kr shared 98.8-99.6% identity. No particular region of variability could be found in either grouping of viruses. All of the CSNV isolates did not show any relationship according to geographical origins and isolation hosts, suggesting no distinct segregation of the CSNV isolates.