• Journal of Microbiology and Biotechnology
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• 제28권5호
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• pp.809-815
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• 2018

Influenza, which is a highly contagious disease caused by the influenza A virus, continues to be a major health concern worldwide. Although the accurate and early diagnosis of influenza virus infection is important for controlling the spread of this disease and rapidly initiating antiviral therapy, the current influenza diagnostic kits are limited by their low sensitivity. In this study, we developed several new influenza nucleoprotein (NP)-specific monoclonal antibodies (mAbs) and compared their sensitivity and specificity of those with commercially available anti-NP mAbs. Three mAbs, designated M24.11, M34.3, and M34.33, exhibited higher reactivities to recombinant NPs and A/Puerto Rico/8/1934 (H1N1) viral lysates compared with the commercial mAbs, as assessed using enzyme-linked immunosorbent assays. M34.3 and M34.33 showed higher reactivities with A/California/04/09 (pandemic H1N1) and A/Philippines/2/82 (H3N2) viral lysates than the commercial mAbs. In contrast, M24.11 had marked reactivity with H3N2 but not with pandemic H1N1. Immunofluorescent confocal microscopy showed that the three mAbs effectively detected the presence of influenza virus in lung tissues of mice infected with A/Puerto Rico/8/1934. These results indicate that the newly developed M34.3 and M34.33 mAbs could be useful for the development of influenza diagnostics.

• 생명과학회지
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• 제11권3호
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• pp.248-253
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• 2001

The outbreak patterns of the internal and external and external patients in the 20 designated hospitals and in 16 health centers were monitored to investigate and the characteristics of the virus isolates were as follows. Two hundreds and thirteen strains of influenza virus were isolated from the oral specimens of 1,686 patients with respiratory disease in Pusan. 1999. Among these isolates, 203 strains were A-type and the rest were B-type. The outbreak patterns for sex and age group were as follows. The male outbreak was similar to the female outbreak: male outbreak, 45.5% and female outbreak, 54.5%. Most of the patients were less than 10 days old. The monthly influenza outbreak was consistent from Jan. to Dec in 1999. The 96 strains from the A-type isolates were A/Sydney/05/97(H3N2)-like, the 107 strains were A/Beijing/262/95(H1N)-like, and all of the 10 B-type isolates were B/Harbin/07/94-like.

• 미생물학회지
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• 제43권3호
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• pp.173-178
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• 2007

2004년과 2005년 동안 부산지역에서 급성호흡기환자로부터 인플루엔자바이러스를 분리 동정하여 분식한 결과, 인플루엔자바이러스분리율은 2004년도는 1869건의 호흡기 검체 중 인플루엔자바이러스 154건 중, A/H3N2형은 77.3%에 해당하는 119건, B형의 경우 35건(22.7%)으로 나타났고 A/H1N1형은 검출되지 않았다. 분리된 인플루엔자바이러스의, 2005년의 경우 1579건의 호흡기 검체 중에서 분리된 인플루엔자바이러스 19건 중, A/H1N1형은 6건(31.6%) 검출되었으며, A/H3N2형은 52.6%에 해당하는 10건, B형의 경우는 3건(15.8%)으로 나타났다. 2005년의 경우 전체적인 인플루엔자바이러스 분리율은 2004년에 비해 떨어졌으나, A/H3N2의 경우 여전히 높은 분리율을 보였으며 2004년에는 전혀 검출되지 않았던 A/H1N1이 B형보다 많이 분리되었다. 부산지역에서 분리된 바이러스의 항원형을 분석한 결과 당해연도 백신주와 동일하거나 유사하였다. 연령별 발생 분포는 0-10세 이하에서 80-90%이상을 차지하였고, 남성에 비해 여성에서 약간 높은 분리율을 나타내었다. 월별 분리율은 2004년도는 4월, 2005년도는 2월이 가장 많이 분리되었다. 인플루엔자바이러스의 대유행 주기에 임박한 현 시점에서 지속적으로 인플루엔자유행예측조사로서 조기 분리한 인플루엔자바이러스 주를 유전자 염기서열을 분석함으로써 신종 인플루엔자바이러스가 출현되는지 적극적인 감시가 필요하다.

• Journal of Microbiology and Biotechnology
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• 제34권3호
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• pp.735-745
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• 2024

Avian influenza is a serious threat to both public health and the poultry industry worldwide. This respiratory virus can be combated by eliciting robust immune responses at the site of infection through mucosal immunization. Recombinant probiotics, specifically lactic acid bacteria, are safe and effective carriers for mucosal vaccines. In this study, we engineered recombinant fusion protein by fusing the hemagglutinin 1 (HA1) subunit of the A/Aquatic bird/Korea/W81/2005 (H5N2) with the Bacillus subtilis poly γ-glutamic acid synthetase A (pgsA) at the surface of Lactobacillus casei (pgsA-HA1/L. casei). Using subcellular fractionation and flow cytometry we confirmed the surface localization of this fusion protein. Mucosal administration of pgsA-HA1/L. casei in mice resulted in significant levels of HA1-specific serum IgG, mucosal IgA and neutralizing antibodies against the H5N2 virus. Additionally, pgsA-HA1/L. casei-induced systemic and local cell-mediated immune responses specific to HA1, as evidenced by an increased number of IFN-γ and IL-4 secreting cells in the spleens and higher levels of IL-4 in the local lymphocyte supernatants. Finally, mice inoculated with pgsA-HA1/L. casei were protected against a 10LD50 dose of the homologous mouse-adapted H5N2 virus. These results suggest that mucosal immunization with L. casei displaying HA1 on its surface could be a potential strategy for developing a mucosal vaccine against other H5 subtype viruses.

• 생명과학회지
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• 제17권3호통권83호
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• pp.340-344
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• 2007

조류인플루엔자바이러스는 사람 및 동물에 상당한 위협을 가하고 있다. 이 연구는 알칼리성 소독용액이 H5N1, H3N2, H6N1, H9N2형 조류인플루엔자바이러스를 불활성화시킬 수 있는지를 조사했다. 알칼리성 소독용액을 생리식염수에 100배 희석한 경우 조류인플루엔자가 MDCK 세포에서 증식하는 것을 완전히 억압하였다. 형광항체법을 이용한 실험에서도 알칼리성소독액을 처리한 H9N2 조류인플루엔자바이러스는 MDCK세포에서 증식이 억제되었고, 알칼리성 소독액을 처리하지 않은 H9N2조류인플루엔자바이러스는 증식이 억제되지 않았다. 이 결과는 알칼리성 소독액은 고병원성 H5N1을 포함을 조류인플루엔자바이러스를 방역하는데 도움이 될 수 있음을 시사한다.

• 대한수의학회지
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• 제38권3호
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• pp.581-588
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• 1998

Eight monoclonal antibodies(MAbs) against bovine coronavirus(BCV) were produced and characterized. Three MAbs(1G9, 4H12, 5C1) specific to the S glycoprotein and two HE glycoprotein-specific MAbs(2A5, 5G4) were found to neutralize the BCV in fluorescence focus neutralization(FFN) test. Two HE-specific MAbs from the neutralizing MAbs inhibited the hemagglutinating activity of the BCV. None of the N protein-specific MAbs(1C1, 5A12, 6H1) neutralized the virus infectivity. Bovine coronavirus and mouse hepatitis virus, which belong to group II coronaviruses, were differentiated from other groups of coronaviruses(porcine transmissible gastroenteritis virus, porcine epidemic diarrhea virus, canine coronavirus) by all MAbs in fluorescence antibody test(FA), but not in FFN test.

• 생약학회지
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• 제31권1호
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• pp.82-86
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• 2000

To evaluate anti-influenza virus activity of 113 specimens of Korean traditional medicine both water and methanol extracts were examined using haemagglutination inhibition test. The water extract from Citrus junos was found to inhibit influenza virus A/Taiwan/l/86(H1N1). The survival rates of virus were determined by in situ cellular enzyme-linked immunosorbent assay. The water extract of Citrus junos was fractionated by chromatographic separating using Amberlite XAD-4, 40% MeOH and 60% MeOH layer had antiviral activity. The half inhibition concentration $(IC_{50})$ of 40% MeOH layer on survival of influenza virus was $MIC>361.5{\mu}g/ml$ and $IC_{50}$ value of fr. 40-4 fractionated from 40% MeOH layer was $677.19{\mu}g/ml$. These results suggested that the fractions of Citus junos have potent anti-influenza A virus activity.

• Animal Bioscience
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• 제35권3호
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• pp.367-376
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• 2022

Objective: The highly pathogenic avian influenza virus (HPAIV) is a threat to the poultry industry as well as the economy and remains a potential source of pandemic infection in humans. Antiviral genes are considered a potential factor for HPAIV resistance. Therefore, in this study, we investigated gene expression related to cytokine-cytokine receptor interactions by comparing resistant and susceptible Ri chicken lines for avian influenza virus infection. Methods: Ri chickens of resistant (Mx/A; BF2/B21) and susceptible (Mx/G; BF2/B13) lines were selected by genotyping the Mx dynamin like GTPase (Mx) and major histocompatibility complex class I antigen BF2 genes. These chickens were then infected with influenza A virus subtype H5N1, and their lung tissues were collected for RNA sequencing. Results: In total, 972 differentially expressed genes (DEGs) were observed between resistant and susceptible Ri chickens, according to the gene ontology and Kyoto encyclopedia of genes and genomes pathways. In particular, DEGs associated with cytokine-cytokine receptor interactions were most abundant. The expression levels of cytokines (interleukin-1β [IL-1β], IL-6, IL-8, and IL-18), chemokines (C-C Motif chemokine ligand 4 [CCL4] and CCL17), interferons (IFN-γ), and IFN-stimulated genes (Mx1, CCL19, 2'-5'-oligoadenylate synthase-like, and protein kinase R) were higher in H5N1-resistant chickens than in H5N1-susceptible chickens. Conclusion: Resistant chickens show stronger immune responses and antiviral activity (cytokines, chemokines, and IFN-stimulated genes) than those of susceptible chickens against HPAIV infection.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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• pp.1719-1727
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• 2014

In the present study, whispovirus immediate early 1 promoter (ie-1) was used to initiate surface expression of the hemagglutinin (HA) protein of Egyptian H5N1 avian influenza virus (AIV) by using the baculovirus expression vector system. The HA gene and whispovirus ie-1 promoter sequence were synthesized as a fused expression cassette (ie1-HA) and successfully cloned into the pFastBac-1 transfer vector. The recombinant vector was transformed into DH10Bac competent cells, and the recombinant bacmid was generated via site-specific transposition. The recombinant bacmid was used for transfection of Spodoptera frugiperda (Sf-9) insect cells to construct the recombinant baculovirus and to induce expression of the HA protein of H5N1 AIV. The recombinant glycoprotein expressed in Sf-9 cells showed hemadsorption activity. Hemagglutination activity was also detected in both extra- and intracellular recombinant HAs. Both the HA and hemadsorption activities were inhibited by reference polyclonal anti-H5 sera. Significant expression of the recombinant protein was observed on the surface of infected insect cells by using immunofluorescence. SDS-PAGE analysis of the expressed protein revealed the presence of a visually distinguishable band of ~63 kDa in size, which was absent in the non-infected cell control. Western blot analysis confirmed that the distinct 63 kDa band corresponded to the recombinant HA glycoprotein of H5N1 AIV. This study reports the successful expression of the HA protein of H5N1 AIV. The expressed protein was displayed on the plasma membrane of infected insect cells under the control of whispovirus ie-1 promoter by using the baculovirus expression vector system.

• Current Optics and Photonics
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• 제4권6호
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• pp.566-570
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• 2020

The present paper computes the refractive indices of different corona viruses (H5N1, H5N2, H9N2, H4N6, FAdV and IBV) through reflectance analysis of a virus solution. The computational analysis indicates that the refractive indices of all viruses are negative at the signal of 412 nm. Further the numerical output shows that the infectious bronchitis viruses (family of novel corona viruses, COVID-19) have higher negative refractive indices as compared to other corona viruses. Finally refractive indices of the family of COVID-19 are investigated with respect to the EID (Electronic infusion Device) concentration of the viruses, showing that the refractive index which ranges from "-0.96725 to -0.999998" corresponds to '0.01 to 10000' EID virus concentration.