• Title/Summary/Keyword: Influenza virus A

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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
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    • v.18 no.4
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    • pp.40.1-40.8
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    • 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.

Investigation of Antibody Titers after Inoculation with Commercial Equine Influenza Vaccines in Thoroughbred Yearlings (Thoroughbred 1세말에서 상업용 말 인플루엔자 백신접종 후 항체역가 추적)

  • Yang, J.H.;Park, Y.S.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.20 no.1
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    • pp.89-96
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    • 2018
  • The object of this study was to evaluate the change of antibody titers on virus strains after inoculation with commercial killed equine influenza (EI) vaccines in horses. Serum antibodies of 20 Thoroughbred yearlings were detected using hemagglutination inhibition test for 41 weeks. Second vaccination is inoculated 4 weeks after the initial vaccination. Most of antibody titers were not increased until 4 weeks after first vaccination. The highest titers were detected 6-10 weeks after vaccination. The titers were decreased slowly and maintained for 16 weeks after inoculation. We could barely detect the antibody 41 weeks after vaccination in most cases. Vaccine anergia were appeared in 3 horses (15%) but it depended on virus strains. A/Equine/La Plata/93(H3N8) strain that induce high and durable antibody responses was the most effective among three strains. This study presents the first comprehensive data on the endurance of antibody titers against EI. Our data also suggests that yearlings should be inoculated three times in order to maintaining optimal antibody titers against EI. We speculate the causes of anergia were vaccine break down or individual specificity. Further research is needed to investigate immunological unresponsiveness. This was the first study on strain of equine vaccine in Korea.

Detection of Avian Influenza-DNA Hybridization Using Wavelength-scanning Surface Plasmon Resonance Biosensor

  • Kim, Shin-Ae;Kim, Sung-June;Lee, Sang-Hun;Park, Tai-Hyun;Byun, Kyung-Min;Kim, Sung-Guk;Shuler, Michael L.
    • Journal of the Optical Society of Korea
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    • v.13 no.3
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    • pp.392-397
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    • 2009
  • We designed a wavelength interrogation-based surface plasmon resonance (SPR) biosensor to detect avian influenza DNA (AI-DNA). Hybridization reactions between target AI-DNA probes and capture probes immobilized on a gold surface were monitored quantitatively by measuring the resonance wavelength in the visible waveband. The experimental results were consistent with numerical calculations. Although the SPR detection technique does not require the DNA to be labeled, we also evaluated fluorescently-labeled targets to verify the hybridization behavior of the AI-DNA. Changes in resonance were found to be linearly proportional to the amount of bound analyte. A wavelength interrogation-type SPR biosensor can be used for rapid measurement and high-throughput detection of highly pathogenic AI viruses.

Etiology and Clinical Features of Viral Lower-respiratory Tract Infections in Children in Winter, 2003 (2003년 겨울철 소아의 바이러스성 하기도 감염증에 대한 원인 및 바이러스별 임상양상)

  • Yun, Byung Ho;Lee, Hee Chul;Chun, Jung Mi;Yoon, So Young;Lee, Woo Gill;Shin, Son Moon
    • Clinical and Experimental Pediatrics
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    • v.48 no.7
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    • pp.723-730
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    • 2005
  • Purpose : This study was performed to characterize the etiology and clinical features of acute viral lower-respiratory tract infections(LRI). Methods : Etiologic agents and clinical features of acute viral LRI were studied from October. 2003 through March. 2004 in hospitalized children with LRI(253 cases) at Samsung Cheil Hospital. The viruses were identified by indirect immunofluorescent method. Medical records of patients with proven viral LRI were reviewed retrospectively. Results : Ninety two cases(36.4%) were confirmed as viral infections. The identified pathogens were respiratory syncytial virus(RSV, 76.0%), adenovirus(ADV, 12.0%), influenza virus type A(INFA, 9.8 %), influenza virus type B(INFB, 1.1%) and parainfluenza virus(PIV, 1.1%). Eight four point eight% of patients were younger than 2 years of age. Clinical diagnosis of LRI were pneumonia(56.5%), bronchiolitis(35.9%), tracheobronchitis(4.3%) and croup(3.3%). The clinical symptoms and signs were cough(98.8%), rhinorrhea(82.6%), fever(70.7%), rale(67.4%), wheezing(29.3%), chest retraction(28.3%) and cyanosis(4.3%). The severe respiratory symptoms and signs were more common in RSV-infected patients, even cyanosis could be observed. Seventeen point four percent of patient had fever of $38.5^{\circ}C$ or higher and their most common etiologic agent was INFA(66.7%). Twenty three point nine percent had fever more than 5 days and common etiologic agent was INFA(77.8%). The elevated WBC count($>14{\times}10^3/{\mu}L$) was in 14.1%, and common etiologic agents were INFA(22.2%) and ADV(18.2%). C-reactive protein(CRP >4.0 mg/dL) was increased in 13.0%, and common in ADV(63.6 %). Increased aspartate aminotransferase(AST)/alanine aminotransferase(ALT) was detected in 10.9%, and the most common etiologic agent was RSV(12.9%). Conclusion : The common agents of acute viral LRI were RSV, ADV and INF, respectively. Because the etiologic agents present variable clinical features, it may be helpful to treat and to evaluate acute viral LRI that we should understand their etiologic variability.

Original Antigenic Sin Response to RNA Viruses and Antiviral Immunity

  • Mee Sook Park;Jin Il Kim;Sehee Park;Ilseob Lee;Man-Seong Park
    • IMMUNE NETWORK
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    • v.16 no.5
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    • pp.261-270
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    • 2016
  • The human immune system has evolved to fight against foreign pathogens. It plays a central role in the body's defense mechanism. However, the immune memory geared to fight off a previously recognized pathogen, tends to remember an original form of the pathogen when a variant form subsequently invades. This has been termed 'original antigenic sin'. This adverse immunological effect can alter vaccine effectiveness and sometimes cause enhanced pathogenicity or additional inflammatory responses, according to the type of pathogen and the circumstances of infection. Here we aim to give a simplified conceptual understanding of virus infection and original antigenic sin by comparing and contrasting the two examples of recurring infections such as influenza and dengue viruses in humans.

Evolution and international transmission of H3N2 canine influenza A viruses from Korea during 2014-2017

  • Chung-Young Lee
    • Journal of Veterinary Science
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    • v.24 no.6
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    • pp.78.1-78.6
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    • 2023
  • Avian-origin H3N2 canine influenza A viruses (CIVs) have become enzootic in China and Korea and have sporadically transmitted to North America, causing multiple epidemics. We isolated six CIVs in Korea from CIV-infected patients during 2014-2017 and conducted whole genome sequencing and phylogenetic analyses. Results revealed that CIVs have circulated and evolved in Korea since the early 2000s and then diversified into a new clade, probably contributing to multiple epidemics in China, the USA, and Canada. Our findings bridge an evolutionary gap for understanding the global transmission of CIVs, emphasizing the significance of continuous monitoring of CIVs.

Detecting of Periodic Fasciculations of Avian Muscles Using Magnetic and Other Multimedia Devices

  • Nakajima, Isao;Tanaka, Sachie;Mitsuhashi, Kokuryo;Hata, Jun-ichi;Nakajima, Tomo
    • Journal of Multimedia Information System
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    • v.6 no.4
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    • pp.293-302
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    • 2019
  • In the past, there was a theory that influenza wasn't transmitted directly from birds but was infected to humans via swains. Recently, molecular level research has progressed, and it was confirmed that the avian influenza virus can directly infected to human lung and intestinal epithelial cells. Three pandemicsin the past 100 years were also infected to humans directly from birds. In view of such scientific background, we are developing a method for screening sick birds by monitoring the physiological characteristics of birds in a contactless manner with sensors. Here, the movement of respiratory muscles and abdominal muscles under autonomic innervation was monitored using a magnet and Hall sensor sewn on the thoracic wall, and other multimedia devices. This paper presents and discusses the results of experiments involving continuous periodic noise discovered during flight experiments with a data logger mounted on a Japanese pheasant from 2012 to 2015. A brief summary is given as the below: 1. Magnet and Hall sensor sewn to the left and right chest walls, bipolar electrocardiograms between the thoracic walls, posterior thoracic air sac pressure, angular velocity sensors sewn on the back and hips, and optical reflection of LEDs (blue and green) from the skin of the hips allow observation of periodic vibrations(fasciculations) in the waves. No such analysis has been reported before. 2. These fasciculations are presumed to be derived from muscle to maintain and control air sac pressure. 3. Since each muscle fiber is spatially Gaussian distributed from the sympathetic nerve, the envelope is assumed to plot a Gaussian curve. 4. Since avian trunk muscles contract periodically at all time, we assume that the sympathetic nerve dominates in their control. 5. The technique of sewing a magnet to the thoracic wall and measuring the strength of the magnetic field with a Hall sensor can be applied to screen for early stage of avian influenza, with a sensor attached to the chicken enclosure.

Critically Ill Patients with Pandemic Influenza A/H1N1 2009 at a Medical Center in Korea

  • Choi, Eun-Young;Huh, Jin-Won;Lim, Chae-Man;Koh, Youn-Suck;Kim, Sung-Han;Choi, Sang-Ho;Kim, Won-Young;Kim, Won;Kim, Mi-Na;Hong, Sang-Bum
    • Tuberculosis and Respiratory Diseases
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    • v.70 no.1
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    • pp.28-35
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    • 2011
  • Background: The aim of the study was to describe the characteristics, treatments, and outcomes of critically ill patients with pandemic Influenza A/H1N1 2009 at a major medical center in Korea. Methods: This retrospective observational study examined critically ill adult patients with pandemic Influenza A/H1N1 2009, who were admitted to the AMC between August and December 2009. Results: 27 patients with confirmed pandemic Influenza A/H1N1 2009 were admitted to the intensive care unit (ICU) at the Asan Medical Center (AMC). The median age (IQR) was 59 years (41~67), and 66.7% of the patients were older than 51 years. A total of 81.5% of the patients had 2 or more co-morbidities. The median time (IQR) from symptom onset to presentation was 2 days (1~4), and the median time from presentation to ICU admission was 0 days (0~1.5). All patients received oseltamivir (300 mg/day) and 13 patients received triple combination therapy (oseltamivir, amantadine, ribavirin). Twelve patients required mechanical ventilation on the first day of ICU admission. A total of 6 patients (22.2%) died within 28 days of admission. The patients who died had significantly higher acute physiology and chronic health evaluation (APACHE) II and sequential organ failure assessment (SOFA) scores at presentation. There were no significant differences in age, co-morbidities, or antiviral regimens between survivors and non-survivors. Conclusion: Critical illness related to pandemic Influenza A/H1N1 2009 was common in elderly patients with chronic co-morbidities. All patients were given high-dose oseltamivir or triple combination antiviral therapy. Nonetheless, patients with critical illnesses associated with pandemic Influenza A/H1N1 2009 had a death rate of 22.2%.

A Study on the Genomic Patterns of SARS coronavirus using Bioinformtaics Techniques (바이오인포매틱스 기법을 활용한 SARS 코로나바이러스의 유전정보 연구)

  • Ahn, Insung;Jeong, Byeong-Jin;Son, Hyeon S.
    • Proceedings of the Korea Contents Association Conference
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    • 2007.11a
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    • pp.522-526
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    • 2007
  • Since newly emerged disease, the Severe Acute Respiratory Syndrome (SARS), spread from Asia to North America and Europe rapidly in 2003, many researchers have tried to determine where the virus came from. In the phylogenetic point of view, SARS virus has been known to be one of the genus Coronavirus, but, the overall conservation of SARS virus sequence was not highly similar to that of known coronaviruses. The natural reservoirs of SARS-CoV are not clearly determined, yet. In the present study, the genomic sequences of SARS-CoV were analyzed by bioinformatics techniques such as multiple sequence alignment and phylogenetic analysis methods as well multivariate statistical analysis. All the calculating processes, including calculations of the relative synonymous codon usage (RSCU) and other genomic parameters using 30,305 coding sequences from the two genera, Coronavirus, and Lentivirus, and one family, Orthomyxoviridae, were performed on SMP cluster in KISTI, Supercomputing Center. As a result, SARS_CoV showed very similar RSCU patterns with feline coronavirus on the both axes of the correspondence analysis, and this result showed more agreeable results with serological results for SARS_CoV than that of phylogenetic result itself. In addition, SARS_CoV, human immunodeficiency virus, and influenza A virus commonly showed the very low RSCU differences among each synonymous codon group, and this low RSCU bias might provide some advantages for them to be transmitted from other species into human beings more successfully. Large-scale genomic analysis using bioinformatics techniques may be useful in genetic epidemiology field effectively.

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Development of Ultra-rapid Multiplex Real-time PCR for the Detection of Genes from Avian Influenza Virus subtype H5N1 (조류인플루엔자 H5N1 바이러스 유전자의 신속 검출을 위한 초고속 다중 실시간 PCR법의 개발)

  • Kim, Eul-Hwan;Lee, Dong-Woo;Han, Sang-Hoon;Lim, Yoon-Kyu;Yoon, Byoung-Su
    • Korean Journal of Veterinary Research
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    • v.47 no.4
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    • pp.399-407
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    • 2007
  • Cause of high lethality and dissemination to human being, new development of rapid method for the detection of highly pathogenic Avian Influenza Virus (AIV) is still necessary. For the detection of AIV subtype H5N1, typical pathogenic AIV, new method to confirm sub-typing of this virus is also needed. For the purpose of ultra-rapid detection and sub-typing of hemagglutinin and neuraminidase of AIV, this study was planned. As the results we could demonstrate an ultra-rapid multiplex real-time PCR (URMRT PCR) for the detection of AIV In this study, the URMRT PCR were optimized with synthesized AIV H5- and AIV Nl-specific DNA templates and GenSpector TMC, which is a semiconductor process technology based real-time PCR system with high frequencies of temperature monitoring. Under eight minutes, the amplifications of two AIV subtype-specific PCR products were successfully and independently detected by 30 cycled ultra-rapid PCR, including melting point analysis, from $1{\times}10^3$ copies of mixed template DNA. The URMRT PCR for the detection of AIV H5N 1 developed in this study could be expected to apply not only detections of different AIVs, but also various pathogens. It was also discussed that this kind of the fastest PCR based detection method could be improved by advance of related technology in near future.