• Journal of Microbiology and Biotechnology
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• 제28권10호
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• pp.1683-1690
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• 2018

Accurate and rapid diagnosis of influenza infection is essential to enable early antiviral treatment and reduce the mortality associated with seasonal and epidemic infections. Immunochromatography is one of the most common methods used for the diagnosis of seasonal human influenza; however, it is less effective in diagnosing pandemic influenza virus. Currently, rapid diagnostic kits for pandemic influenza virus rely on the detection of nucleoprotein (NP) or hemagglutinin (HA). NP detection shows higher specificity and is more sensitive than HA detection. In this study, we time-dependently screened expression conditions, and herein report optimal conditions for the expression of recombinant nucleoprotein (rNP), which was 48 h after infection. In addition, we report the use of the expressed rNP in a rapid influenza diagnostic test (SGT i-flex Influenza A&B Test). We constructed expression vectors that synthesized rNP (antigen) of influenza A and B in insect cells (Sf9 cells), employed the purified rNP to the immunoassay test kit, and clearly distinguished NPs of influenza A and influenza B using this rapid influenza diagnostic kit. This approach may improve the development of rapid test kits for influenza using NP.

• Journal of Veterinary Science
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• 제21권2호
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• pp.24.1-24.11
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• 2020

The pandemic of avian influenza viruses (AIVs) in Asia has caused enormous economic loss in poultry industry and human health threat, especially clade 2.3.4.4 H5 and H7 subtypes in recent years. The endemic chicken H6 virus in Taiwan has also brought about human and dog infections. Since wild waterfowls is the major AIV reservoir, it is important to monitor the diversified subtypes in wildfowl flocks in early stage to prevent viral reassortment and transmission. To develop a more efficient and sensitive approach is a key issue in epidemic control. In this study, we integrate multiplex reverse transcription recombinase polymerase amplification (RT-RPA) and capillary electrophoresis (CE) for high-throughput detection and differentiation of AIVs in wild waterfowls in Taiwan. Four viral genes were detected simultaneously, including nucleoprotein (NP) gene of all AIVs, hemagglutinin (HA) gene of clade 2.3.4.4 H5, H6 and H7 subtypes. The detection limit of the developed detection system could achieve as low as one copy number for each of the four viral gene targets. Sixty wild waterfowl field samples were tested and all of the four gene signals were unambiguously identified within 6 h, including the initial sample processing and the final CE data analysis. The results indicated that multiplex RT-RPA combined with CE was an excellent alternative for instant simultaneous AIV detection and subtype differentiation. The high efficiency and sensitivity of the proposed method could greatly assist in wild bird monitoring and epidemic control of poultry.

• Journal of the Korean Data and Information Science Society
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• 제27권4호
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• pp.1075-1081
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• 2016

The big data analysis has received much attention from the researchers working in various fields because the big data has a great potential in detecting or predicting future events such as epidemic outbreaks and changes in stock prices. Reflecting the current popularity of big data analysis, many authors have proposed methods tracking influenza epidemics based on internet-based information. The recently proposed 'autoregressive model using Google (ARGO) model' (Yang et al., 2015) is one of those influenza tracking models that harness search queries from Google as well as the reports from the Centers for Disease Control (CDC), and appears to outperform the existing method such as 'Google Flu Trends (GFT)'. Although the ARGO predicts well the outbreaks of influenza, this study demonstrates that a classical seasonal autoregressive integrated moving average (SARIMA) model can outperform the ARGO. The SARIMA model incorporates more accurate seasonality of the past influenza activities and takes less input variables into account. Our findings show that the SARIMA model is a functional tool for monitoring influenza epidemics.

• 한국시뮬레이션학회논문지
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• 제23권4호
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• pp.31-39
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• 2014

계절적 유행성 독감은 매년 전 세계적으로 300만-500만 명이 감염되어 25만-50만 명이 사망에 이르는 무서운 질병이다. 유행성 독감에 대한 통제를 강화하기 위해 독감의 유행을 실시간으로 감시하기 위한 연구들이 제안되고 있다. 우리나라의 질병관리본부는(CDC)는 인플루엔자에 대한 임상 자료를 1주 단위로 발표하고 있으며 질병의 유행과 1-2주 정도의 보고 시차가 존재한다. 조기에 독감의 유행을 감지하기 위해 비임상적 자료(뉴스 리포트, 소셜 미디어)의 검색 정보를 활용하여 유행성 독감 발생을 효과적으로 대비하기 위한 연구들이 최근 이루어지고 있다. 비임상적 자료의 수집은 적은 비용으로 거의 실시간으로 이루어질 수 있는 이점이 있다. 본 연구는 네이버 검색엔진이 제공하는 PC와 모바일 키워드 정보를 활용하여 우리나라의 유행성 독감 활동을 감지하는 회귀모형을 개발하고자 한다. 이를 위해 문헌연구를 통하여 인플루엔자 의사분율(ILI)과 높은 상관성을 가질 것으로 예상되는 키워드를 20개 선정하고 키워드와 ILI와의 관계를 로지스틱 회귀모형과 다중회귀모형으로 가정하고 ILI를 예측하였다. 모형적합성 측면에서 다중회귀모형이 로지스틱모형보다 우수하였으며 모바일-기반 회귀모형이 PC-기반 회귀모형보다 ILI 퍼센티지를 추정하는데 우월한 결과를 보이고 있다.

• Journal of Microbiology and Biotechnology
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• 제24권5호
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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.

• Clinical and Experimental Pediatrics
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• 제55권12호
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• pp.470-473
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• 2012

Purpose: The purpose of this prospective case-control study was to survey the detection rate of respiratory viruses in children with Kawasaki disease (KD) by using multiplex reverse transcriptase-polymerase chain reaction (RT-PCR), and to investigate the clinical implications of the prevalence of respiratory viruses during the acute phase of KD. Methods: RT-PCR assays were carried out to screen for the presence of respiratory syncytial virus A and B, adenovirus, rhinovirus, parainfluenza viruses 1 to 4, influenza virus A and B, metapneumovirus, bocavirus, coronavirus OC43/229E and NL63, and enterovirus in nasopharyngeal secretions of 55 KD patients and 78 control subjects. Results: Virus detection rates in KD patients and control subjects were 32.7% and 30.8%, respectively (P=0.811). However, there was no significant association between the presence of any of the 15 viruses and the incidence of KD. Comparisons between the 18 patients with positive RT-PCR results and the other 37 KD patients revealed no significant differences in terms of clinical findings (including the prevalence of incomplete presentation of the disease) and coronary artery diameter. Conclusion: A positive RT-PCR for currently epidemic respiratory viruses should not be used as an evidence against the diagnosis of KD. These viruses were not associated with the incomplete presentation of KD and coronary artery dilatation.