• Title/Summary/Keyword: Influenza A viruses

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Sero-prevalence of swine influenza virus (H1N1, H3N2) In Korea (국내의 돼지 인플루엔자 바이러스(H1N1, H3N2)의 혈청학적 조사)

  • Yoon, Jai-soon;Park, Bong-kyun;Han, Jeong-hee
    • Korean Journal of Veterinary Research
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    • v.47 no.3
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    • pp.273-279
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    • 2007
  • Swine influenza is an acute, infectious respiratory disease caused by type A influenza viruses in pigs. In the previous studies, serological surveys have indicated the presence of H3N2 swine influenza virus (SIV) since 1995 in Korea. And the percentage of the antibody-positive rate was 39.12% in the survey determining the prevalence of H1N1 SIV antibodies in 2002. The purpose of this study was therefore to investigate the sero-prevalence of SIV regard to the age of the pig and the season between June 2004 and May 2005. In this study, a total of 932 sera were used. These sera were randomly selected from blood samples, which were submitted to Department of Veterinary Pathology, Kangwon National University and Department of Veterinary Virology, Seoul National University from June 2004 to May 2005. These sera have been tested by ELISA test kit (IDEXX Lab, USA) for the SIV H3N2, H1N1 respectively. SAS version 9.1 was used for the statistical analysis based on the age of the pig and the season. The overall sero-prevalence of the antibody against H3N2 SIV was 20.82% (194/932). The overall sero-prevalence of the antibody against H1N1 SIV was 37.23% (347/932). The overall dual sero-prevalence of the antibody against H3N2 and H1N1 SIV was 10.62% (99/932). H3N2 has significant difference in statistically regarding the age of the pig and the season (p<0.0001). H1N1 has significant difference in statistically regarding the age of the pig (p<0.0001) but has not significant difference in statistically regarding the season (p=0.5882).

Integrated Rotary Genetic Analysis Microsystem for Influenza A Virus Detection

  • Jung, Jae Hwan;Park, Byung Hyun;Choi, Seok Jin;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.88-89
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    • 2013
  • A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

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Ginseng, the natural effectual antiviral: Protective effects of Korean Red Ginseng against viral infection

  • Im, Kyungtaek;Kim, Jisu;Min, Hyeyoung
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.309-314
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    • 2016
  • Korean Red Ginseng (KRG) is a heat-processed ginseng developed by the repeated steaming and air-drying of fresh ginseng. Compared with fresh ginseng, KRG has been shown to possess greater pharmacological activities and stability because of changes that occur in its chemical constituents during the steaming process. In addition to anticancer, anti-inflammatory, and immune-modulatory activities, KRG and its purified components have also been shown to possess protective effects against microbial infections. Here, we summarize the current knowledge on the properties of KRG and its components on infections with human pathogenic viruses such as respiratory syncytial virus, rhinovirus, influenza virus, human immunodeficiency virus, human herpes virus, hepatitis virus, norovirus, rotavirus, enterovirus, and coxsackievirus. Additionally, the therapeutic potential of KRG as an antiviral and vaccine adjuvant is discussed.

Needs of Biosecurity and Protocols for the Environmental Management of Carcasses Burial (가축매몰지 환경관리에 있어 차단방역의 필요 및 절차)

  • Cho, Ho-Seong;Kim, Geonha
    • Journal of Korean Society on Water Environment
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    • v.28 no.2
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    • pp.305-312
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    • 2012
  • Avian influenza (AI) and foot and mouth disease (FMD) are two main contagious pathogenic viruses causing massive mortality burial, as burial is a primary measure to quaranteen the causative viruse(s). Biosecurity is a set of preventive measures designed to reduce the risk of propagation of infectious diseases. Main objectives of this paper were to discuss the needs of biosecurity and develop protocol outlines for environmental management of burial sites. Pathological characteristics of contagious viruses should be considered during environmental management practices. Current practice prescribes to minimize the potential for on-farm pollution and the spread of infectious diseases, policy makers should understand robust knowledge regarding biosecurity to make informed decisions on future legislation.

Establishment of optimal disinfection condition of weak acid hypochlorous solution for prevention of avian influenza and foot-and-mouth disease virus transmission (조류 인플루엔자와 구제역 바이러스 차단방역을 위한 미산성 차아염소산수의 소독 조건)

  • Kim, Jin-Yoon;Yun, Dong-Sik;Lee, Haw-Yong;Jeong, Woo-Seog;Park, Seung-Chun
    • Korean Journal of Veterinary Research
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    • v.59 no.2
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    • pp.101-104
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    • 2019
  • This study examined the disinfection conditions (exposure time, 0-30 min; exposure temperature, $4^{\circ}C-65^{\circ}C$) of hypochlorous acid water (HOCl) in automobile disinfection equipment. The study tested poliovirus type 1 (PV1), low pathogenic avian influenza virus (AIV, H9N2), and foot and mouth disease virus (FMDV, O type). As a result, the PV1 and FMD viruses were inactivated easily (virus titer 4 log value) by HOCl (> 100 ppm) but the AIV required higher exposure temperatures (> $55^{\circ}C$). In conclusion, the exposure temperature and time are important factors in deactivating AIV and FMDV.

Improving siRNA design targeting nucleoprotein gene as antiviral against the Indonesian H5N1 virus

  • Hartawan, Risza;Pujianto, Dwi Ari;Dharmayanti, Ni Luh Putu Indi;Soebandrio, Amin
    • Journal of Veterinary Science
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    • v.23 no.2
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    • pp.24.1-24.10
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    • 2022
  • Background: Small interfering RNA technology has been considered a prospective alternative antiviral treatment using gene silencing against influenza viruses with high mutations rates. On the other hand, there are no reports on its effectiveness against the highly pathogenic avian influenza H5N1 virus isolated from Indonesia. Objectives: The main objective of this study was to improve the siRNA design based on the nucleoprotein gene (siRNA-NP) for the Indonesian H5N1 virus. Methods: The effectiveness of these siRNA-NPs (NP672, NP1433, and NP1469) was analyzed in vitro in Marbin-Darby canine kidney cells. Results: The siRNA-NP672 caused the largest decrease in viral production and gene expression at 24, 48, and 72 h post-infection compared to the other siRNA-NPs. Moreover, three serial passages of the H5N1 virus in the presence of siRNA-NP672 did not induce any mutations within the nucleoprotein gene. Conclusions: These findings suggest that siRNA-NP672 can provide better protection against the Indonesian strain of the H5N1 virus.

One step multiplex RT-PCR preventing DNA carryover contamination for differential diagnosis of swine influenza viruses (DNA 교차 오염 방지 기능을 가진 돼지 인플루엔자바이러스 감별진단용 one-step multiplex RT-PCR 진단법)

  • Kim, Hee-Jung;Kim, Eun-Mi;Shin, Yeun-Kyung;Song, Jae-Young;Kim, Seong-Hee;Lee, Kyoung-Ki;Lee, Myoung-Heon;Kim, Young-Hwa;Park, Jun-Cheol;Yeo, Sang-Geon;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.37 no.4
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    • pp.263-271
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    • 2014
  • In this study, we developed a cost and time saving one-step multiplex RT-PCR for the simultaneous detection and differentiation of swine influenza viruses (SIV) and 2009 pandemic influenza H1N1 virus (pH1N1). The one-step multiplex RT-PCR using four sets of primer was confirmed to be capable of detection of all SIV subtypes and differential diagnosis of major SIV subtype H1, H3 and pH1N1 on individual or mixed viral culture samples. The sensitivity of the multiplex RT-PCR was determined to be at least $2^{-6}$ $HA/25{\mu}L$ of the presented SIVs, providing sufficient efficacy for a routine SIV monitoring in diagnostic laboratories. In addition, compared with the conventional RT-PCR methods that cannot avoid the carryover DNA contamination, the developed RT-PCR applied with the uracil DNA glycosylase (UNG) system was proven to prevent a false positive reaction by carryover contamination of the pre-amplified DNA. In conclusion, the one-step RT-PCR with UNG system could be applicable to detect and differentiate of SIV from the viral cultures without worry of carryover DNA contamination in clinical laboratories.

The Efficacy of Rapid Antigen Tests for Detection of Seasonal Influenza Virus (계절 인플루엔자 진단에서 신속항원검사의 유용성)

  • Lee, Jun Yeol;Lee, Sol;Kim, Han Sung;Kim, Kwang Nam
    • Pediatric Infection and Vaccine
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    • v.24 no.1
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    • pp.31-36
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    • 2017
  • Purpose: This study aimed to examine the accuracy of rapid influenza diagnostic tests (RIDT) in children with an influenza-like illness and to evaluate factors associated with greater accuracy. Methods: Pediatric patients, who visited Hallym University Sacred Heart Hospital with an influenza-like illness between June 2011 and May 2016, were enrolled in this study. We tested 798 samples using a real-time polymerase chain reaction (PCR) for respiratory viruses and compared the results with rapid influenza tests. Results: In comparison with the results of the multiplex PCR, the positive agreement rates of RIDT for influenza A and B virus were 75.7% and 60.0%, respectively. The performance of RIDT varied according to days after fever onset. The positive agreement rates of RIDT for influenza A and B tests, performed within 4 days of fever onset, were 77.6% and 73.2%, but the rates for tests performed more than 5 days after fever onset were 66.7% and 21.4%, respectively. Conclusions: The RIDT is a quick and simple aid to diagnosis, but is less sensitive than the labeled sensitivity. Moreover, test performance varied according to days after fever onset. Test specimens for RIDT should be collected as soon as possible after the onset of symptoms (less than 4 days).

Paper-Based Neuraminidase Assay Sensor for Detection of Influenza Viruses (인플루엔자 바이러스 검출을 위한 종이 기반 neuraminidase 효소 활성 평가 센서 개발)

  • Hwang, Cheol-hwan;Jeong, Seong-Geun;Park, Han-Kyu;Lee, Chang-Soo;Kim, Yun-Gon
    • Korean Chemical Engineering Research
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    • v.54 no.3
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    • pp.380-386
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    • 2016
  • In this study, we described a paper-based neuraminidase assay sensor (PNAS) which can be applied to detect the infection by influenza viruses. The PNAS was designed and manufactured to quantitatively identify the levels of neuraminidase in the sample, which is based on colorimetric analysis using the X-Neu5Ac substrate. The limit of detection of the PNAS was determined as 0.004 U/mL of neuraminidase. According to the amount of neuraminidase in human serum, the PNAS could monitor the enzyme activity with a good linearity ($R^2$ > 0.99). In addition, the initial performance of the PNAS has been maintained up to 70 days in the $4^{\circ}C$. Finally, we demonstrated whether the Michaelis-Menten kinetics is applied to the PNAS, which can show the reliability of the enzyme reactions. The kinetic studies indicated that the PNAS provides the good condition for enzyme reactions ($K_m=8.327{\times}10^{-3}M$), but they were performed on paper chip nonetheless. The paper-based neuraminidase assay sensor may be useful in a wide range of rapid and safe detection of influenza virus.

Novel Phage Display-Derived H5N1-Specific scFvs with Potential Use in Rapid Avian Flu Diagnosis

  • Wu, Jie;Zeng, Xian-Qiao;Zhang, Hong-Bin;Ni, Han-Zhong;Pei, Lei;Zou, Li-Rong;Liang, Li-Jun;Zhang, Xin;Lin, Jin-Yan;Ke, Chang-Wen
    • Journal of Microbiology and Biotechnology
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    • v.24 no.5
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    • pp.704-713
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    • 2014
  • The highly pathogenic avian influenza A (HPAI) viruses of the H5N1 subtype infect poultry and have also been spreading to humans. Although new antiviral drugs and vaccinations can be effective, rapid detection would be more efficient to control the outbreak of infections. In this study, a phage-display library was applied to select antibody fragments for HPAI strain A/Hubei/1/2010. As a result, three clones were selected and sequenced. A hemagglutinin inhibition assay of the three scFvs revealed that none exhibited hemagglutination inhibition activity towards the H5N1 virus, yet they showed a higher binding affinity for several HPAI H5N1 strains compared with other influenza viruses. An ELISA confirmed that the HA protein was the target of the scFvs, and the results of a protein structure simulation showed that all the selected scFvs bound to the HA2 subunit of the HA protein. In conclusion, the three selected scFVs could be useful for developing a specific detection tool for the surveillance of HPAI epidemic strains.