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Isolation of subgroup J avian leukosis virus in Korea
Haan-Woo Sung,Jae-Hong Kim,Sanjay Reddy,Aly Fadly 대한수의학회 2002 Journal of Veterinary Science Vol.3 No.2
Two subgroup J avian leukosis viurses (ALVs) wereisolated from broiler breeder flocks,in which myeloidleukosis had occurred.The isolates could be classifiedas subgroup JALV by the positive reaction in polymerasechain reaction (PCR) with primers specific for subgroupJ ALV. Two isolates replicated in chicken embryofibroblast (CEF) cells from the alv6 chicken line inwhich cells are resistant to subgroup A and E ALVs.In in vitro serum neutralization tests with othersubgroup ALVs including ADOL-Hc1,the prototype ofsubgroup J ALVs isolated in the United States ofAmerica, two isolates were partially neutralized byantibody to ADOL-Hc1, indicating that Korean isolatesand ADOL-Hc1 may be antigenically related, but notidentical.When the PCR was done with a primer pairdesigned to amplify genes of E element and longterminal repeat of proviral DNA, the PCR productsize of one isolate (KOAL-PET) was smaller than thatof ADOL-Hc1, suggesting that some sequences inthese regions are deleted.
성환우,김재홍,Sung, Haan-woo,Kim, Jae-hong 대한수의학회 2000 大韓獸醫學會誌 Vol.40 No.1
Duck viral hepatitis is an acute, highly infectious viral disease of young dacklings aged from two days to three weeks. The significant lesion associated with the disease was enlarged liver including necrotic foci and numerous hemorrhagic spots. We have isolated five strains of duck hepatitis virus (DHV) from field cases showing about 20% mortality with a sign of opisthotonos. When a-day-old ducklings were intramuscularly inoculated with one of the isolates, 92% of the birds were died within 5 days. We attempted to develop an attenuated strain of duck hepatitis virus (DHV) using one of the isolates by serial chicken embryo passages. The propagation of DHV in chicken embryos was carried 140 passages. The virus titer increased gradually from the $21^{st}$ through the $50^{th}$ passage, but there was no significant increase of virus titer in subsequent passages after then. Through the serial passages, the virulence of the virus for chicken embryos was gradually increased but decreased for ducklings. The pathogenicity of the virus for ducklings was preserved up to the $21^{st}$ passage but disappeared at the $50^{th}$passage. An attenuated Korean isolate which was passaged 140 times in chicken embryos gave good protection in ducklings against both challenge infection to a Korean virulent strain and to a DHV-DRL strain, a type 1 reference strain of DHV, which indicated that the Korean isolates could be classified as DHV type 1. And the above results suggest that an attenuated Korean isolate can be used for developing a live DHV vaccine.
성환우,이수정,Sung, Haan Woo,Lee, Su Jeong 대한수의학회 2005 大韓獸醫學會誌 Vol.45 No.4
Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.
Moon, Hyun-Woo,Sung, Haan Woo,Park, Jeongho,Kwon, Hyuk Moo The Korean Society of Veterinary Science 2021 大韓獸醫學會誌 Vol.61 No.2
To evaluate avian hepatitis E virus (aHEV) as an RNA vaccine platform, ORF2 of aHEV was replaced by heterologous genes, such as eGFP and HA-tag, in aHEV infectious cDNA clones. eGFP and HA-tag replicons were expressed in LMH cells. To confirm expression of the heterologous protein, ORF2 was replaced with the antigenic S1 gene of IBV. The IBVS1 replicon was expressed in LMH cells. To our knowledge, this is the first investigation showing the potential as a RNA vaccine platform using an aHEV. In the future, it may be used in the development of RNA vaccines against various pathogens.