• 대한수의학회지
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• 제59권2호
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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.

• Journal of Veterinary Science
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• 제24권5호
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• pp.73.1-73.16
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• 2023

Background: Highly pathogenic avian influenza virus (HPAIV) is considered a global threat to both human health and the poultry industry. MicroRNAs (miRNA) can modulate the immune system by affecting gene expression patterns in HPAIV-infected chickens. Objectives: To gain further insights into the role of miRNAs in immune responses against H5N1 infection, as well as the development of strategies for breeding disease-resistant chickens, we characterized miRNA expression patterns in tracheal tissues from H5N1-infected Ri chickens. Methods: miRNAs expression was analyzed from two H5N1-infected Ri chicken lines using small RNA sequencing. The target genes of differentially expressed (DE) miRNAs were predicted using miRDB. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis were then conducted. Furthermore, using quantitative real-time polymerase chain reaction, we validated the expression levels of DE miRNAs (miR-22-3p, miR-146b-3p, miR27b-3p, miR-128-3p, miR-2188-5p, miR-451, miR-205a, miR-203a, miR-21-3p, and miR-200a3p) from all comparisons and their immune-related target genes. Results: A total of 53 miRNAs were significantly expressed in the infection samples of the resistant compared to the susceptible line. Network analyses between the DE miRNAs and target genes revealed that DE miRNAs may regulate the expression of target genes involved in the transforming growth factor-beta, mitogen-activated protein kinase, and Toll-like receptor signaling pathways, all of which are related to influenza A virus progression. Conclusions: Collectively, our results provided novel insights into the miRNA expression patterns of tracheal tissues from H5N1-infected Ri chickens. More importantly, our findings offer insights into the relationship between miRNA and immune-related target genes and the role of miRNA in HPAIV infections in chickens.

• 미생물학회지
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• 제47권1호
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• pp.30-37
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• 2011

조류 인플루엔자 바이러스(AIV) 아형을 ultra-time PCR법(UPCR)을 이용하여 초스피드로 진단할 수 있는 방법을 고안하였다. 표적 대상의 프라이머는 AIV H5N1 아형의 hemagglutinin(HA) 유전자 중 가장 상보성이 높은 133 bp의 부위를 선택하였고, 실험의 안전을 위하여 인공합성의 방법으로 제작하였다. 압타머와 결합한 molecular beacon 기반 Mini-Opticon Q-PCR 기기를 사용한 UPCR법으로, 총 UPCR 반응액의 양을 10 ${\mu}l$으로, UPCR과 용융온도 분석시간을 15분 이내로 매우 짧게 단축시켰다. 민감도 측정에서 최소의 주형인 5분자의 HA 유전자만으로 정확히 AIV의 특이적 133 bp를 합성하였다. UPCR로 디자인된 이 PCR은 AIV 아형의 진단에 적용될 수 있을 뿐 아니라, UPCR이 기반되는 진단을 이용하여 다른 병원체에도 널리 적용 될 수 있을 것으로 기대된다.

• 대한수의학회지
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• 제42권1호
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• pp.73-80
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• 2002

An enzyme-linked immunosorbent assay (ELISA) using purified hemagglutinin of swine influenza virus (H1N1) as antigen was developed for detection of antibody to avian influenza virus (AIV). The sensitivity and specificity of a developed and commercial available ELISA kits were compared with those of agar gel precipitation (AGP) test and hemagglutination inhibition (HI) test using sera collected from chickens under condition of field exposure. The concentration of antigen, serum dilution and concentration of enzyme-conjugated secondary antibody in developed ELISA (S-ELISA) were 0.5ug/100ul, 1:200 and 0.03ug/100ul, respectively. The correlation coefficients between S-ELISA and commercial ELISA and HI titers were 0.419 and 0.533, respectively. A significant correlation (p < 0.01) was not found between HI and ELISA titers. The S-ELISA was found to be as more sensitive and specific than the AGP test, showing 86.8% sensitivity and 85.3% specificity. It is suggested that the ELISA using the SIV as antigen may be useful method as an investigating tool for AIV serological surveillance.

• 대한수의학회지
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• 제47권4호
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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.

• 한국가금학회지
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• 제31권2호
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• pp.109-118
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• 2004

Avian influenza(AI) is an epizootic disease of variable severity caused by type A influenza viruses of the orthomyxovirus group. Chickens were the most frequently affected avian species with AI viruses. There were many outbreaks of fowl plague, now known as highly pathogenic AI(HP AI), throughout the world since Perroncito described the fowl plague in 1978 in Italy. In recent years HPAI viruses of different serotypes such as H5, H7 and H9 has been isolated from humans on several occasions either related with outbreak of HPAI in birds or not. In 1997, one of the most noteworthy events in AI history was the human mortality with H5N1 HPAI virus infection in Hong Kong. Six persons of total 18 persons with clinical signs of influenza were died. Recently the human cases with mortality related with HP AI outbreaks in poultry industry has been increased such as outbreaks of HP AI throughout Asia countries including Korea, Japan, China, Vietnam, Thailand and others in 2003. Although these outbreaks revealed the capable of spreading from birds to human, the capability for transmission between people was not clear. Therefore, this report will review the possibility of HP AI infection in human associated with HPAI outbreak in poultry industry.

• 한국동물위생학회지
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• 제40권1호
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• pp.15-20
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• 2017

A two-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was designed for the rapid visual detection of the M gene of all subtypes of avian influenza virus (AIV) and the H5 gene of the H5 subtype of highly pathogenic AIV (HPAIV). The reaction carried out in two tubes in a single step at $58^{\circ}C$ for 40 min, and the assay results could be visually detected by using hydroxynaphthol blue dye. Using M or H5 gene-specific primers, the assay successfully detected all subtypes or H5 subtypes of AIVs, including the Korean representative H5N1 and H5N8 HPAIVs. The detection limit of the assay was approximately $10^{2.0}$ $EID_{50}/reaction$ for the M and H5 genes of H5N1 HPAIV, respectively, and was more sensitive than that of previously reported RT-LAMP and comparable to that of real-time RT-PCR. These results suggest that the present RT-LAMP assay, with its high specificity, sensitivity, and simplicity, will be a useful diagnostic tool for surveillance of currently circulating H5 HPAIVs and other subtypes of AIV in bird population, even in under-equipped laboratories.

• Tuberculosis and Respiratory Diseases
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• 제70권1호
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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%.

• Journal of Microbiology and Biotechnology
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• 제26권11호
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• pp.1972-1982
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• 2016

Influenza A virus is a single-stranded RNA virus with a genome of negative polarity. Owing to the antigenic diversity and cross concrete shift, an immense number of novel strains have developed astronomically over the years. The present work deals with the codon utilization partialness among five different influenza A viruses isolated from human hosts. All the subtypes showed the homogeneous pattern of nucleotide utilization with a little variation in their utilization frequencies. A lower bias in codon utilization was observed in all the subtypes as reflected by higher magnitudes of an efficacious number of codons. Dinucleotide analysis showed very low CpG utilization and a high predilection of A/T-ending codons. The H5N1 subtype showed noticeable deviation from the rest. Codon pair context analysis showed remarkable depletion of NNC-GNN and NNT-ANN contexts. The findings alluded towards GC-compositional partialness playing a vital role, which is reflected in the consequential positive correlation between the GC contents at different codon positions. Untangling the codon utilization profile would significantly contribute to identifying novel drug targets that will pacify the search for antivirals against this virus.

• Journal of Veterinary Science
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• 제19권6호
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• pp.817-826
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• 2018

The bursa of Fabricius (BF) is a central humoral immune organ unique to birds. Four bursal peptides (BP-I, BP-II, BP-III, and BP-IV) have been isolated and identified from the BF. In this study, the immunoadjuvant activities of BPs I to IV were examined in mice immunized with H9N2 avian influenza virus (AIV) vaccine. The results suggested that BP-I effectively enhanced cell-mediated immune responses, increased the secretion of Th1 (interferon gamma)- and Th2 (interleukin-4)-type cytokines, and induced an improved cytotoxic T-lymphocyte (CTL) response to the H9N2 virus. BP-II mainly elevated specific antibody production, especially neutralizing antibodies, and increased Th1- and Th2-type cytokine secretion. BP-III had no significant effect on antibody production or cell-mediated immune responses compared to those in the control group. A strong immune response at both the humoral and cellular levels was induced by BP-IV. Furthermore, a virus challenge experiment followed by H&E staining revealed that BP-I and BP-II promoted removal of the virus and conferred protection in mouse lungs. BP-IV significantly reduced viral titers and histopathological changes and contributed to protection against H9N2 AIV challenge in mouse lungs. This study further elucidated the immunoadjuvant activities of BPs I to IV, providing a novel insight into immunoadjuvants for use in vaccine design.