• Journal of Veterinary Science
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• 제21권4호
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• pp.56.1-56.13
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• 2020

Background: The live bird market (LBM) plays an important role in the dynamic evolution of the avian influenza H5N1 virus. Objectives: The main objective of this study was to monitor the genetic diversity of the H5N1 viruses in LBMs in Indonesia. Methods: Therefore, the disease surveillance was conducted in the area of Banten, West Java, Central Java, East Java, and Jakarta Province, Indonesia from 2014 to 2019. Subsequently, the genetic characterization of the H5N1 viruses was performed by sequencing all 8 segments of the viral genome. Results: As a result, the H5N1 viruses were detected in most of LBMs in both bird' cloacal and environmental samples, in which about 35% of all samples were positive for influenza A and, subsequently, about 52% of these samples were positive for H5 subtyping. Based on the genetic analyses of 14 viruses isolated from LBMs, genetic diversities of the H5N1 viruses were identified including clades 2.1.3 and 2.3.2 as typical predominant groups as well as reassortant viruses between these 2 clades. Conclusions: As a consequence, zoonotic transmission to humans in the market could be occurred from the exposure of infected birds and/or contaminated environments. Moreover, new virus variants could emerge from the LBM environment. Therefore, improving pandemic preparedness raised great concerns related to the zoonotic aspect of new influenza variants because of its high adaptivity and efficiency for human infection.

• Clinical and Experimental Pediatrics
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• 제56권4호
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• pp.165-175
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• 2013

Purpose: There was a global increase in the prevalence of oseltamivir-resistant influenza viruses during the 2007-2008 influenza season. This study was conducted to investigate the occurrence and characteristics of oseltamivir-resistant influenza viruses during the 2007-2008 and 2008-2009 influenza seasons among patients who were treated with oseltamivir (group A) and those that did not receive oseltamivir (group B). Methods: A prospective study was conducted on 321 pediatric patients who were hospitalized because of influenza during the 2007-2008 and 2008-2009 influenza seasons. Drug resistance tests were conducted on influenza viruses isolated from 91 patients. Results: There was no significant difference between the clinical characteristics of groups A and B during both seasons. Influenza A/H1N1, isolated from both groups A and B during the 2007-2008 and 2008-2009 periods, was not resistant to zanamivir. However, phenotypic analysis of the virus revealed a high oseltamivir $IC_{50}$ range and that H275Y substitution of the neuraminidase (NA) gene and partial variation of the hemagglutinin (HA) gene did not affect its antigenicity to the HA vaccine even though group A had a shorter hospitalization duration and fewer lower respiratory tract complications than group B. In addition, there was no significant difference in the clinical manifestations between oseltamivir-susceptible and oseltamivir-resistant strains of influenza A/H1N1. Conclusion: Establishment of guidelines to efficiently treat influenza with oseltamivir, a commonly used drug for treating influenza in Korean pediatric patients, and a treatment strategy with a new therapeutic agent is required.

• Clinical and Experimental Pediatrics
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• 제52권8호
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• pp.862-868
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• 2009

Since its identification in April 2009, a swine-origin H1N1 influenza A virus (S-OIV) which is a reassortment of gene segments from both North American triple-reassortant and Eurasian swine influenza has been widely spread among humans in unexpected rapidity. To date, each gene segment of the 2009 influenza A (H1N1) outbreak viruses have shown high (99.9%) neucleotide sequence identity. As of July 6, 94,512 people have been infected in 122 countries, of whom 429 have died with an overall case-fatality rate of <0.5%. Most confirmed cases of S-OIV infection have been characterized by self-limited, uncomplicated febrile respiratory illness and 38% of cases have also included vomiting or diarrhea. Standard plus droplet precautions should be adhered to at all times. Tests on S-OIV have indicated that current new H1N1 viruses are sensitive to neuraminidase inhibitors (oseltamivir). However, current less virulent S-OIV may evolve into a pathogenic strain or acquire antiviral resistance, potentially with more severe clinical consequences. Efforts to control these outbreaks would be based on our understanding of novel S-OIV and previous influenza pandemics.

• 한국환경보건학회지
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• 제37권4호
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• pp.306-314
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• 2011

Objectives: Respiratory virus infections are the most common disease among all ages in all parts of the world and occur through airborne transmission. The purpose of this study was to detect and quantitate human respiratory viruses in residential environments. Methods: Air samples were collected from the residential space of apartments in the Seoul/Gyeonggi-do area. The samples were collected from indoor and outdoor air. Among respiratory viruses, influenza A virus, influenza B virus, parainfluenza virus, metapneumovirus, respiratory syncytial virus, and adenovirus were investigated by multiplex polymerase chain reaction. Among the virus-positive samples, we performed adenovirus quantification by real-time polymerase chain reaction. Results: Virus detection rates were 44.0%, 3.8%, 3.4%, and 17.3% in spring, summer, autumn, and winter, respectively. The virus detection rate was higher in winter and spring than in summer and autumn. Adenovirus was most commonly detected, followed by influenza A virus and parainfluenza virus. Virus distribution was not significantly different between indoor and outdoor environments. Conclusions: Although virus concentrations were not high in residential environments, residents in houses with detected viruses may have an increased risk of exposure to airborne respiratory viruses, especially in winter and spring.

• 한국가금학회지
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• 제46권3호
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• pp.173-183
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• 2019

조류인플루엔자 바이러스 H7 subtype에 속하는 바이러스 중 일부는 가금류에 감염할 경우 고병원성이 발휘된다. 또 H7 아형 AIV중 일부는 사람에 감염하여 사망 등을 유발할 수도 있다. 본 연구는 야생조류로부터 분리된 H7 아형 조류인플루엔자 바이러스 6주(H7N7 아형 4주, H7N1 아형 2주)를 대상으로 8개 유전자 분절 전체의 염기서열을 분석하여 병원성, 사람 감염 가능성 등 그 특성을 조사하였다. 계통유전학적 분석결과, 국내에서 분리된 H7 아형 분리주들은 8개 유전자(HA, NA, PB2, PB1, PA, NP, M, NS) 모두 Eurasian lineage로 분류되었으나, Eurasian lineage 내에서도 각기 다른 sublineage로 분류되어 유전적 다양성이 있는 것으로 분석되었다. 한국 분리주 6주는 HA 단백질 분절부위 아미노산은 두 종류(PEIPKGR 및 PELPKGR)의 motif를 가지고 있었으나, 모두 저병원성 바이러스 특성을 가지고 있었다. 숙주세포 결합 특이성과 관련 있는 HA 단백질 receptor-binding site를 분석한 결과, 한국 분리주 모두는 사람 세포 수용체 결합특이성보다는 조류 세포 수용체 결합 특이성을 가지는 것으로 나타났다. 사람 감염 가능성을 높게 하는 부위에서의 아미노산 치환(PB2 단백질의 E627K 및 PB1단백질의 I368V)도 나타나지 않았고, 또한 NA stalk region에서의 결손도 관찰되지 않았다. 이상의 결과를 미루어 볼 때 한국 야생조류에서 분리된 H7 아형 6주 모두는 저병원성 바이러스로 최근 중국에서 사람 감염이 나타나고 있는 H7N9 바이러스와는 유전적으로 다른 계열의 바이러스인 것으로 판단된다.

• Genomics & Informatics
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• 제18권1호
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• pp.5.1-5.5
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• 2020

Highly pathogenic avian influenza (HPAI) viruses have caused severe respiratory disease and death in poultry and human beings. Although most of the avian influenza viruses (AIVs) are of low pathogenicity and cause mild infections in birds, some subtypes including hemagglutinin H5 and H7 subtype cause HPAI. Therefore, sensitive and accurate subtyping of AIV is important to prepare and prevent for the spread of HPAI. Next-generation sequencing (NGS) can analyze the full-length sequence information of entire AIV genome at once, so this technology is becoming a more common in detecting AIVs and predicting subtypes. However, an analysis pipeline of NGS-based AIV sequencing data, including AIV subtyping, has not yet been established. Here, in order to support the pre-processing of NGS data and its interpretation, we developed a user-friendly tool, named prediction of avian influenza virus subtype (PAIVS). PAIVS has multiple functions that support the pre-processing of NGS data, reference-guided AIV subtyping, de novo assembly, variant calling and identifying the closest full-length sequences by BLAST, and provide the graphical summary to the end users.

• Genomics & Informatics
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• 제20권4호
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• pp.46.1-46.7
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• 2022

Influenza A virus (IAV) is the most widespread pathogen causing human respiratory infections. Although polymerase chain reaction (PCR)-based methods are currently the most commonly used tools for IAV detection, PCR is not ideal for point-of-care testing. In this study, we aimed to develop a more rapid and sensitive method than PCR-based tools to detect IAV using loop-mediated isothermal amplification (LAMP) technology. We designed reverse-transcriptional (RT)-LAMP primers targeting the hemagglutinin gene. RNAs from reference H1N1 and H3N2 showed specific RT-LAMP signals with the designed primers. We optimized the reaction conditions and developed universal reaction conditions for both LAMP assays. Under these conditions, the detection limit was 50 copies for both RT-LAMP assays. There was no non-specific signal to 19 non-IAV respiratory viruses, such as influenza B virus, coronaviruses, and respiratory syncytial viruses. Regarding the reaction time, a positive signal was detected within 25 min after starting the reaction. In conclusion, our RT-LAMP assay has high sensitivity and specificity for the detection of the H1 and H3 subtypes, making it suitable for point-of-care IAV testing.

• IMMUNE NETWORK
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• 제20권4호
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• pp.32.1-32.15
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• 2020

Influenza virus is the major cause of seasonal and pandemic flu. Currently, oseltamivir, a potent and selective inhibitor of neuraminidase of influenza A and B viruses, is the drug of choice for treating patients with influenza virus infection. However, recent emergence of oseltamivir-resistant influenza viruses has limited its efficacy. Morin hydrate (3,5,7,2',4'-pentahydroxyflavone) is a flavonoid isolated from Morus alba L. It has antioxidant, anti-inflammatory, neuroprotective, and anticancer effects partly by the inhibition of the NF-κB signaling pathway. However, its effects on influenza virus have not been studied. We evaluated the antiviral activity of morin hydrate against influenza A/Puerto Rico/8/1934 (A/PR/8; H1N1) and oseltamivir-resistant A/PR/8 influenza viruses in vitro. To determine its mode of action, we carried out time course experiments, and time of addition, hemolysis inhibition, and hemagglutination assays. The effects of the co-administration of morin hydrate and oseltamivir were assessed using the murine model of A/PR/8 infection. We found that morin hydrate reduced hemagglutination by A/PR/8 in vitro. It alleviated the symptoms of A/PR/8-infection, and reduced the levels of pro-inflammatory cytokines and chemokines, such as TNF-α and CCL2, in infected mice. Co-administration of morin hydrate and oseltamivir phosphate reduced the virus titers and attenuated pulmonary inflammation. Our results suggest that morin hydrate exhibits antiviral activity by inhibiting the entry of the virus.

• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.997-1006
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• 2018

As shown during the 2009 pandemic H1N1 (A(H1N1)pdm09) outbreak, egg-based influenza vaccine production technology is insufficient to meet global demands during an influenza pandemic. Therefore, there is a need to adapt cell culture-derived vaccine technology using suspended cell lines for more rapid and larger-scale vaccine production. In this study, we attempted to generate a high-growth influenza vaccine strain in MDCK cells using an A/Puerto/8/1934 (H1N1) vaccine seed strain. Following 48 serial passages with four rounds of virus plaque purification in MDCK cells, we were able to select several MDCK-adapted plaques that could grow over $10^8PFU/ml$. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells ($PB1_{D153N}$, $M1_{A137T}$, and $NS1_{N176S}$). In addition, we confirmed that in the context of the high-growth A/PR/8/34 backbone, A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), and A/environment/Korea/deltaW150/2006 (H5N1) also showed significantly enhanced growth properties (more than $10^7PFU/ml$) in both attached- and suspended-MDCK cells compared with each representative virus and the original PR8 vaccine strain. Taken together, this study demonstrates the feasibility of a cell culture-derived approach to produce seed viruses for influenza vaccines that are cheap and can be grown promptly and vigorously as a substitute for egg-based vaccines. Thus, our results suggest that MDCK cell-based vaccine production is a feasible option for producing large-scale vaccines in case of pandemic outbreaks.

• 한국축산식품학회지
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• 제28권4호
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• pp.512-516
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• 2008

To search for anit-H5N1 influenza virus agent, the anti-viral activity of methanol and aqueous extracts from thirty medicinal plants were examined in this study. The plant material (30 g) was extracted with methanol (300 mL) for 24 hr at room temperature. Methanol extracts were filtered and evaporated, then freeze-dried. Aqueous extracts were prepared with dried plant material (30 g) and hot distilled water (300 mL). After 3 hr, the aqueous extracts were filtered and evaporated, then lyophilized. Extracts prepared from different plants were tested the antiviral activity against influenza viruses [A/vietnam/1194/04 (H5N1)-NIBRG-14] using the hemagglutination (HA) assay. Among the test plants, Asarum sieboldii was found to be a potent inhibitor of H5N1 influenza virus in MDCK cell culture. Virus titers were 7 log, whereas with methanol extract of Asarum sieboldii for 48 hr titers were 3 log, indicating that methanol extract of Asarum sieboldii inhibited the H5N1 influenza viruses from the infected cells.