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한타바이러스 혈청형 특이 Primer를 이용한 Nested RT - PCR 방법으로 5가지 혈청형 한타바이러스에 감염된 햄스터 조직에서 바이러스 검출
주용규,이호왕 대한바이러스학회 1997 Journal of Bacteriology and Virology Vol.27 No.1
We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.
$1996{\sim}1998$년 유고슬라비아에서 실시한 한탄바이러스 불활화 백신($Hantavax^{TM}$)의 면역효과에 대한 야외시험
주용규,우영대,안창남,김훈,장양석,박승철,김민자,이은실,이호왕,Chu, Yong-Kyu,Gligic, Anna,Tomanovic, Snezana,Bozovjc, Boyana,Obradovic, Mirceta,Woo, Young-Dae,An, Chang-Nam,Kim, Hun,Jiang, Yang-Seok,Park, Seung-Chul,Kim, Min-Ja 대한미생물학회 1999 Journal of Bacteriology and Virology Vol.28 No.1
In Yugoslavia, hemorrhagic fever with renal syndrome (HFRS) is one of the important national health problem, but no vaccine has been used to prevent HFRS. Since first HFRS case in 1952, sporadic cases of HFRS occurred every year and over 4,000 registered cases with $1{\sim}16%$ mortality so far. We performed a prospective, randomized double-blind placebo-controlled trial to evaluate the effectiveness of $Hantavax^{TM}$ against HFRS in 3,900 healthy adults living in the endemic areas of Yugoslavia. 1,900 people were given 0.5 ml of Hantavax subcutaneously twice at one month interval and a booster shot at one year after. For controls other 2,000 healthy people were given 0.5 ml of physiolosical saline as a placebo. We investigated HFRS cases in both the vaccinated and nonvaccinated groups by monitoring the program for patient registration in the areas from 1996 to 1998, and the effect of vaccine was analyzed epidemiologically. No confirmed case of HFRS was observed among 1,900 Hantavax vaccinees, while 20 confirmed cases were observed among 2,000 nonvaccinated control group. There were no remarkable side effects among the vaccinees either locally or in general after inoculation of the vaccine. The Hantavax vaccine showed statistically significant protective efficacy against HFRS among Yugoslavian people.
한타바이러스 호왕균주의 M , S 유전자 절편의 염기서열 및 분자생물학적 특성
주용규,이호왕 대한바이러스학회 1997 Journal of Bacteriology and Virology Vol.27 No.1
Hantaan virus Howang strain which isolated from the blood of severe case of Korean hemorrhagic fever is more virulent than HTN 76/118 and showed different RFLP from partial PCR amplifed M genome segment to established Hantaan serotype viruses. We have determined the nucleotide sequence of the M and S genome segments and compared to HTN 76/118. The M and S segment of Howang strain has 3615 and 1696 nucleotides long, respectively. The M segment sequence of Howang strain is one mucleotide shorter than HTN 76/118. The sequence data of Howang strain shows 93.5% homology to HTN 76/118. One long open reading frame, which strats from 41nt. To 3448nt. Of the M segment and from 37nt. To 1326nt. Of the S segment, exist to on complementary sense of the virus genome. There are no significant difference between HTN 76/118 and Howang strain on hydrophobicity of deduced polypeptides, but has slight difference on secondary structure.
한탄바이러스 면역혈청에 대한 각종 동물 FITC-conjugated Anti-immunoglobulin G의 교차반응
주용규,이광래,이호왕 대한바이러스학회 1985 Journal of Bacteriology and Virology Vol.15 No.1
After primary reaction of Hantaan virus antigen and antiserum, different FITC-conjugated anti-IgG were applied to antigen-antibody complex for observation of cross reactivity between different origin of IgG. Immune sera preparated by immunizing S.D. rats, Balb/c mice and rabbits with Hantaan virus and human convalescent serum from Hemorrhagic fever with renal syndrome (HFRS) patients were used to test the cross reaction to different animal FITC-conjugated anti-immunoglobulin G, using the indirect immuno- fluorescent antibody different (IFA) technique. The results were as follows: 1. Human convalescent serum from HFRS patient which was combined with Hantaan virus antigen cross reacted to eight FITC-conjugated animal anti-immunoglobulin G sera. 2. Rat immune serum which was combined with Hantaan virus antigen did not cross react to FITC- goat anti-rabbit IgG for 1 hour reaction, but for 4 hour reaction it cross reacted to FITC-goat anti- rabbit IgG serum. 3. Mouse immune serum which was combined with Hantaan virus antigen did not cross react to FITC- goat anti-rahbit IgG. 4. Rabbit immune serum which was combined with Hantaan virus antigen did not cross react to FITC rabbit anti-human IgG, anti-rat IgG and anti-mouse IgG.
韓國型 出血熱 IX . 병원체 (病原體) 한탄 (漢灘)바이러스를 접종한 (接種) Albino마우스에서의 免疫글로불린 形成에 關한 硏究
주용규(朱容奎),백락주(白樂柱),성인화(成仁華),이용주(李容周),이호왕(李鎬汪) 대한바이러스학회 1982 Journal of Bacteriology and Virology Vol.12 No.1
In this study albino mice were inoculated with different strains of Hantaan virus intramuscularly which were propagated in different host cells and found that strain of the virus propagated in Vero cells produced good antibody response in the mice. 1. Immunoglobulin G against Hantaan virus antigen was started to appear on 5th day after inoculation of the virus, reached maximun on 2nd to 3rd week and kept for 2 weeks and then declined slowly. 2. Immunoglobulin M to the virus was started to appear on 9th day after inoculation and reached maximum on 2nd to 3rd week and kept for 2 weeks and then declined slowly and disappeared around 5th week after inaculation of the virus. 3. Immunoglobulin A to the virus was not demonstrated at all in the mice.
주용규(Yong Gyu Joo),송대용(Dae Yong Song),구형모(Hyung Mo Goo),이호왕(Ho Wang Lee) 대한바이러스학회 1999 Journal of Bacteriology and Virology Vol.29 No.3
Hantaan virus (HTNV), the etiologic agent of hemorrhagic fever with renal syndrome (HFRS), belongs to the genus Hantavirus, and has three single negative stranded RNA genome segments. HTNV strain Howang isolated from the blood of severe case of Korean HFRS is more virulent than HTNV 76/118 and the M and S genome segments' nucleotide sequence of Howang strain showed 93.5% and 94% homology to each segment of HTNV 76/118. We have obtained 6533 nucleotides long sequence of the L genome segment of Howang strain using reverse transcriptase in conjunction with PCR amplification and compared to other hantaviruses. The messenger sense of the L segment contains one long single long open reading frame of 2151 amino acids, which encodes a deduced RNA dependent RNA polymerase of 246.4 kDa caculated molecular weight protein. The nucleotide sequence of the L segment of Howang strain shows 93%, 74%, 66%, 65% homology to HTNV 76/118, Seoul virus 80/39, Puumala virus hallnas B1 and Sin Nombre virus, respectively. The amino acid sequence of the L segment of Howang strain shows 99%, 85%, 68%, 68% homology to HTNV 76/118, Seoul virus 80/39, Puumala virus Hallnas B1 and Sin Nombre virus, respectively.