인도네시아와 태국에서 채집된 Bandicota indica 폐장조직에서 한타바이러스 분리

우영대,주용규,이호왕 대한바이러스학회 1998 Journal of Bacteriology and Virology Vol.28 No.2

Various hantaviruses were isolated from HFRS patients and various rodent species, in many parts of the world. Bandicotas were captured at Yogyakarta, east region of Sumatura island, Indonesia; and 4 rodents species including Bandicotas were captured at Chiang Rai in Thailand during 1995. Sera were collected from captured bandicotas and other rodent spicies were screened for antibody test against Hantaan (HTN), Seoul (SEO), Puumala (PUU) and Sin Nombre (SN) viruses by immunofluoresence antibody assay (IFA). Hantavirus antigen in lung tissues were tested by IFA. Among 55 captured Bandicota indica in Indonesia, 14 (25.5%) were antibody positive against HTN, SEO, PUU and SN virus. Hantavirus antigen were detected from 5 (9.0%) out of 55 lungs tested. Among 34 captured Bandicota indica in Thailand, 9 (26.5%) were antibody positive against HTN, SEO, PUU and SN virus. Among 34 lungs tissues of Bandicota indica examined, 3 (8.8%) were antigen positive. In other rodent species, antibody positive against Hantaviruses of Rattus rattus, Rattus losea and Mus cervicolor were 4/62 (6.5%), 5/25 (20%), 1/1 (100%), respectively. But no one has antigen in their lung tissues. Antigen positive lungs suspension were inoculated into vero E6 cells for virus isolation and 4 viruses were isolated from Indonesian Badicota and 3 viruses from Thailand.

신증후출혈열 환자의 혈청학적 및 분자생물학적 진단 검사법 비교

우영대,문희주,배형준 대한의생명과학회 2000 Journal of biomedical laboratory sciences Vol.6 No.2

우리나라에서 발생하고 있는 급성 출혈성 질환인 신증후출혈열의 원인 바이러스는 Family Bunyaviridae의 Genus Hantavirus에 속하는 한탄과 서울바이러스에 의하여 발생되고 있다. 본연구에서는 신증후출혈열로 의뢰된 환자에서 한탄바이러스에 대한 항체가를 간접면역형광항체법(indirect immunofluorescent antibody technique, IFAT), 면역효소측정법(enzyme-linked immunosorbent assay, ELISA) (IgG, IgM), 고비중입자응집반응(high density composite particle agglutination, HDPA) 및 플라크감소중화시험(plaque reduction neutrafization test, PRNT)등으로 비교 측정하였고, 신증후출혈열환자로 확진된 15명의 한타바이러스 혈청형을 PRNT와 혈청형 특이 역전사 효소 중합효소연쇄반응(nested reverse transcriptase polymerase chain reaction, nested RT-PCR)으로 확인하였다. 신증후출혈열로 의뢰된 환자에서의 한탄바이러스에 대한 IFAT, ELISA(IgG,IgM),HDPA 그리고 PRNT 비교에서 형광항체, ELISA IgG, 응집항체 및 중화항체는 8명 모두 높게 나타났으며, ELISA IgM은 5명에서는 현저히 높은 항체를 보유하고 있었다. 신증후출혈열 환자 15명에서는 높은 형광항체와 중화항체 역가를 나타내었고, 15명 중 12명은 한탄바이러스, 2명은 서울바이러스에 대한 높은 중화항체를 갖고 있었으며, 1명은 두 바이러스에 대하여 동일한 항체 역가를 나타내었으며, 혈청형 특이 primer를 사용한 nested RT-PCR에서는 15명 중 3명과 1명만이 한탄바이러스와 서울바이러스 primer에 대해 RNA가 검출되었다. The etiologic agents of haemorrhagic fever with renal syndrome(HFRS) in Korea are Hantaan and Seoul virus in the genus Hantavirus, family Bunyaviridae. Antibody titers of sera from HFRS patients against Hantaan virus were measured by immunofluorescent antibody technique(IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA) and plaque reduction neutralization test (PRNT). PRNT and nested reverse transcriptase polymerase chain reaction (nested RT-PCR) was used for serotypic differentiation of Hantaviruses against Hantaan and Seoul virus. Eight doubtful HFRS patients showed higher fluorescent, IgG ELISA, agglutination and neutralizing antibody titer by IFAT, ELISA IgG, HDPA and PRNT, respectively. Five out of them showed high IgM antibody titer by IgM capture ELISA against Hantaan virus, remarkably. Fifteen HFRS patients showed higher fluorescent antibody titer by IFAT. In PRNT, 12 out of them showed high neutralizing antibody titer aginst HTNV, 2 against SEOV and 1 against both viruses. In nested RT-PCR using serotype specific-primer, 3 out of them showed positive against HTNV and 1 against SEOV.

Survery of Seroconversion Rates against Hantavirus in Laboratory Rodents

우영대,--,--,--,-- THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 2003 Journal of biomedical laboratory sciences Vol.9 No.2

Hantavirus ale rodent-borne RNA virus that belongs to the family Bunyaviridae. Those viruses persistently infect a variety of rodents, and are transmitted by aerosols of their urine, feces and saliva. Antibody titers of sera obtained from normal laboratory rodents against hantavirus were investigated by indirect immunofluorscence antibody technique (IFA). Seroconversion rates of normal laboratory rodents showed higher in rats than that from hamster and mongolian (M), gerbil. Theses rates of normal laboratory rodents also showed higher in titers against puumala virus (PUUV) than in hantaan (HTNV) and seoul virus (SEOV). We are concerned about infections caused by hantaviruses, especially by PUUV, occurred in laboratory rodents.

A Study on Protection of Maternal Antibody against Hantavirus in Rats

우영대,--,--,--,--,-- 대한의생명과학회 2005 Biomedical Science Letters Vol.11 No.1

The etiologic agents of haemorragic fever with ranal syndrom (HFRS) in Korea are Hantaan and Seoul virus in the genus Hantavirus, family Bunyaviridae. In order to elucidate the role of maternal immunity to Hantavirus infection in rats, the protective effect of the maternal antibody were studies by using rats experimentally infected with Seoul virus strain HR80-39. Antibody titers of sera and viral antigen against Seoul virus were investigated by indirect immunofluorscence antibody technique (IFA). The dam sera had IFA antibody titers ranging from 1:128 to 1:1,024 after parturition. In fetuses, IFA antibody titers ranged from 1:16 to 1:64 just after birth, increased to peak titers ranged from 1:256 to 1:1,024 in the 2nd week after birth. Challenged newborn rats had IFA antibody titers ranging from 1:64 to 1:1,024 after inoculation. No viral antigen was detected in lungs or other organs of the newborn rats. The maternal antibody to Seoul virus was transferred prenatally through placenta and postnatally via colostrum from immune dams to their offspring. These results demonstrated that maternal antibody to Seoul virus was quite effective in protecting newborn rats against same virus infection.

1994년~2000년 사이에 혈청학적으로 진단된 신증후출혈열의 혈청 역학적 조사

우영대(Young-Dae Woo),박상욱(Sang-Wook Park),강재명(Jae-Myung Kang),우준희(Jun-Hee Woo),이호왕(Ho-Wang Lee) 대한미생물학회 2001 Journal of Bacteriology and Virology Vol.31 No.2

신증후출혈열 백신 한타박스 접종자에서의 면역반응 및 항체지속 기간에 관한 연구

우영대,주용규,이호왕 대한감염학회 1998 감염 Vol.30 No.4

목 적 : 우리나라에서 한탄바이러스와 서울 바이러스에 의하여 발생하는 신증후출혈열을 예방하기 위하여 1990년 개발된 한타박스는 현재 상용화되어 있다. 한타박스가 시중에서 상용화되어 사용된 역사가 짧으므로, 예방접종자에서 항체지속기간에 관한 연구는 미흡한 실정이다. 본 연구에서는 신증후출혈열 예방백신인 한타박스를 접종한 사람들에서의 항체 특히 중화항체양전율과 항체지속기간을 조사하고자 하였다. 방 법 : 아산생명과학연구소에 근무하는 61명의 연구원들을 대상으로 신증후출혈열 예방백신인 한타박스를 한달간격으로 2회 기본접종하였다. 예방백신을 접종하기 전 대상자들이 혈청을 채취하여 한탄바이러스와 서울바이러스에 대한 항체음성임을 형광항체법과 중화항체법으로 확인하였다. 한타박스를 기본접종한 한달 후 및 일년 후 그리고 기본 접종한지 일년이 경과하여 1차 부스터 접종을 한 한달 후 및 2년여가 경과한 후와 2차 부스터 접종 한달 후의 백신접종자에서의 항체양성율 및 항체가를 간접면역형광항체법(IFA), 고비중입자응집반응(HDPA) 및 플라크감소 중화시험(PRNT) 등으로 비교 측정하여 배신접종자에서의 항체양성율, 형광항체가 및 중화항체가와 면역지속기간을 조사하였다. 결 과 : 1) 신증후출혈열 백신인 한타박스를 2회 기본접종을 하고 한달 후 혈청을 채취한 21명에서의 IFA, HDPA와 PRNT방법에 의한 항체양성율은 각각 20/21 (95.2%), 19/21 (90.5%), 14/21 (66.7%)이었으며, 평균 항체역가는 각각 262,248,120이었다. 2) 기본접종 1면 후 혈청을 채취한 40명에서의 IFA, HDPA와 PRNT방법에 의한 항체양성율은 저하되어 각각 25/40 (62.5%), 18/40 (45.0%) 그리고 9/40 (22.5%)이었으며 평균 항체역가는 각각 92,41,24.5 이었다. 3) 부스터접종이 효과를 조사하여 본 결과 기본접종 1년후 채혈한 14명에서의 IFA와 PRNT방법에 의한 항체양성율은 7/14 (50.0%), 3/14 (21.4%)이었으며 평균 항체역가는 각각 100.6, 47.1 이었다. 1차 부스터 접종 한 14명중 1개월후 채혈한 8명의 IFA와 PRNT방법에 의한 항체양성율은 8/8(100%), 8/8(100%)이었으며 평균 항체역가는 각각 852,182.5 이었다. 1차 부스터 접종한 20개월후 채혈된 12명의 항체양성율은 11/12 (91.7%), 9/12 (75.0%)이었으며 평균 항체역가는 각각 296, 33.3 이었고, 2차 부스터 접종한 3개월 후 채혈한 7명의 항체양성을율은 7/7 (100%), 6/7(85.7%)으로 항체 평균역가는 각각 548,46 이었따. 기본접종 1년후 1차부스터접종을 실시한 1개월후 채혈한 8명의 항체역가와 2차부스터접종을 실시한 한달후 채혈한 7명의 항체역가를 형광항체법과 중화항체법으로 조사한 결과 뚜렷한 항체역가의 증가를 나타내는 기왕성면역반응을 볼 수 있었다. 결 론 : 백신접종자에서 한탄바이러스에 대한 항체를 높게 유지하기 위해서는 2회의 기본접종과 기본접종 1년후 추가접종이 필요하며, 추가접종후 최소한 2년간은 항체가 지속적으로 유지된다는 사실을 증명하였다. Background : Hantavax™ was developed and has been used for prevention of hemorrhagic fever with renal syndrome caused by Hantaan and Seoul virus since 1990 in Korea. Since Hantavax™ has short usage history, the duration of antibodies persistancy in vaccinees was not well studied. Methods : Hantavax™ was inoculated to 61 people who work at Asan Institue for Sciences twice subcutaneously at one month interval according to the manufacturer's recommendation. Antibody titers of vaccinees were measured at 1∼4 months or 1 year after primary basic vaccination and a month, 1∼2 years after booster vaccination by indirect immunofluorescent assay (IFA), High density particle agglutination assay (HDPA), and plaque reduction neutralization test (PRNT). Results : Serconversion of twentyone vaccinees on 1∼4 months after primary vaccination were 20/21 (95.2%), 19/21 (90.5%) and 14/21 (66.7%) whose geometric mean antibody titers were 262,248,120; Forty vaccinees on one year after primary vaccination were 25/40 (62.5%), 18/40 (45.0%), 9/40 (22.5%) whose geometric mean antibody titers were 90,56,24 by IFA, HDPA, PRNT, respectively. Seroconversion of eight vaccinees on one month after booster vaccination were 11/12 (91.7%), 9/12 (75.0%) whose mean antibody titers were 296,33, and seven vaccinees on 3 months after second booster vaccination were 7/7 (100%), 6/7 (85.7%) whose mean antibody titers were 54,946 by IFA, PRNT, respectively. Conclusion : The boost vaccination is necessary at 12 months after primary basic vaccination of Hantavax™ for maintaining high level of antibodies against of Hantaan virus, and antibodies persist at least two years.

신증후출혈열 백신의 면역혈청학적 연구

우영대(Young Dae Woo),주용규(Yong Gyu Joo),백락주(Rak Joo Baek),이호왕(Ho Wang Lee) 대한바이러스학회 2000 Journal of Bacteriology and Virology Vol.30 No.1

Since Hantavax, formalin inactivated Hantaan virus vaccine (10,240 ELISA units/ml), has been developed in 1990 to prevent against haemorrhagic fever with renal syndrome (HFRS) caused by Hantaan or Seoul virus, it has been commercially available in Korea. Twenty-one healthy people were booster shot once and twice after prirnary basic vaccination with Hantavax. Seroconversion rates were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA), and plaque reduction neutralization test (PRNT). Seroconversion rates of 21 vaccinees at one year after primary basic vaccination were 52.3%, 95.2%, 0.0%, 47.6%, and 28.6%, and 13 vaccinees at one month after 1st booster vaccination were 100%, 100%, 30.7%, 100% and 100% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates declined slightly by twenty months, and they were 84.6%, 92.3%, 0.0%, 84.6% and 69.2% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates of 9 vaccinees at three months after 2nd booster vaccination were 100%, 100%, 0.0%, 100%, and 88.9%, and 16 vaccinees at one year after the 2nd booster vaccination were 87.5%, 93.8%, 0.0%, 87.5% and 81.3% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Based on the above result Hantavax has proved a vigorous anamnestic response after the 1st and the 2nd booster vaccination and has persisted higher fluorescence, agglutination and neutralizing antibody titers in vaccinees.

1994~1998년 한국에서 채집한 설치류의 한타바이러스 감염에 대한 혈청역학적 조사

우영대(Young-Dae Woo),주용규(Yong-Kyu Chu),Longzhu Cui,이호왕(Ho-Wang Lee) 대한미생물학회 2003 Journal of Bacteriology and Virology Vol.33 No.1

신증후출혈열 환자의 혈청학적 및 분자생물학적 진단 검사법 비교

우영대(Young-Dae Woo),문희주(Hi-Joo Moon),배형준(Hyung-Joon Bae) 대한의생명과학회 2000 Biomedical Science Letters Vol.6 No.2

우리 나라에서 발생하고 있는 급성 출혈성 질환인 신증후출혈열의 원인 바이러스는 Family Bunyaviridae의 Genus Hantavirus에 속하는 한탄과 서울바이러스에 의하여 발생되고 있다. 본 연구에서는 신증후출혈열로 의뢰된 환자에서 한탄바이러스에 대한 항체가를 간접면역형광항체법 (indirect immunofluorescent antibody technique, IFAT), 면 역효소측정법 (enzyme-linked immunosorbent assay, ELISA) (IgG, IgM), 고비중입자응집반응 (high density composite particle agglutination, HDPA) 및 플라크감소중화시험 (plaque reduction neutralization test, PRNT) 등으로 비교 측정하였고, 신증후출혈열 환자로 확진된 15명의 한타바이러스 혈청형을 PRNT와 혈청형 특이 역전사 효소 중합효소연쇄반응 (nested reverse transcriptase polymerase chain reaction, nested RT-PCR)으로 확인하였다. 신증후출혈열로 의뢰된 환자에서의 한탄바이러스에 대한 IFAT, ELISA (IgG, IgM), HDPA 그리 고 PRNT 비교에서 형광항체, ELISA IgG, 응집항체 및 중화항체는 8명 모두 높게 나타났으며, ELISA IgM은 5명에서는 현저히 높은 항체를 보유하고 있었다. 신증후출혈열 환자 15명에서는 높은 형광항체와 중화항체 역가를 나타내었고, 15명 중 12명은 한탄바이러스, 2명은 서울바이러스에 대한 높은 중화항체를 갖고 있었으며, 1명은 두 바이러스에 대하여 통일한 항체 역가를 나타내었으며, 혈청형 특이 primer를 사용한 nested RT-PCR에서는 15명 중 3명과 1명만이 한탄바이러스와 서울바이러스 primer에 대해 RNA가 검출되었다. The etiologic agents of haemorrhagic fever with renal syndrome (HFRS) in Korea are Hantaan and Seoul virus in the genus Hantavirus, family Bunyaviridae. Antibody titers of sera from HFRS patients against Hantaan virus were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA) and plaque reduction neutralization test (PRNT). PRNT and nested reverse transcriptase polymerase chain reaction (nested RT-PCR) was used for serotypic differentiation of Hantaviruses against Hantaan and Seoul virus. Eight doubtful HFRS patients showed higher fluorescent, IgG ELISA, agglutination and neutralizing antibody titer by IFAT, ELISA IgG, HDPA and PRNT, respectively. Five out of them showed high IgM antibody titer by IgM capture ELISA against Hantaan virus, remarkably. Fifteen HFRS patients showed higher fluorescent antibody titer by IFAT. In PRNT, 12 out of them showed high neutralizing antibody titer aginst HTNV, 2 against SEOV and 1 against both viruses. In nested RT-PCR using serotype specific-primer, 3 out of them showed positive against HTNV and 1 against SEOV.

Salmonella typhimurium에서 MudJ(Km.lac) 오페론 융합을 이용한 삼투유도유전자의 발현

주성관,우영대,허연주,안정선,박용근 한국미생물학회 1991 미생물학회지 Vol.29 No.4

MudJ(Km.lac) operon fusions were used in the identification of osmotic-inducible genetic(osi) loci in Salmonella typhimurium. Expression of osi::lacZ(osi5001, 5027) genes were dramatically induced 39-189 fold when the osmolarity was increased. Seven osm::lacZ genes were constituvely expressed under both low and high osmotic strength. The osi5001::lacZ fusion strains showed the enhanced osmotolerance and the reduced expression of the osi5001::lacZ in the presence of 1mM proline or betaine as osmoprotectants. Four osmotic inducible genetic loci were mapped into 36 (YK531), 44 (YK504), 57 (YK501) and 84 (YK528) map unit by testing the cotransduction frequency.