보르나병 바이러스 항체검출을 위한 연구 -세 가지 혈청진단법의 비교-

이두식,Lee, Du-sik The Korean Society of Veterinary Science 1992 大韓獸醫學會誌 Vol.32 No.1

Borna disease(BD) virus 특이항체검출에 대한 세 가지 혈청진단법(간접형광항체법, 세포효소면역반응법, 혈청중화시험)의 정확도를 비교하기 위해 BD virus를 실험적으로 감염시킨 273수의 토끼의 혈청으로 시험하였다. 혈청중 간접형광항체법에 의하여 판정된 123혈청들은 모두 세포효소면역반응법에 의해서 양성으로 판정되었으나 혈청중화시험법에 의해서는 단지 27혈청에서만 양성으로 판정되었다. 혈청중화시험법은 간접형광항체법과 세포효소면역반응법에 비하여 민감도가 훨씬 낮게 나타나 BD virus의 혈청학적 연구에는 간접형광항체법 및 세포효소면역반응법의 활용도가 높을 것으로 생각된다. To determin the accuracy of serological methods in detecting Borna-disease(BD) viral antibodies, 273 experimentally infected rabbit sera were compared by using indirect immunofluorescence antibody test(IFA), serum neutralization test(SN) and enzyme-linked immunosorbent assay(ELISA). One hundred twenty-three serum samples had BD viral antibodies detected by IFA. CELISA antibodies to BD virus were also present in the same one hundred twenty-three serum samples. However, neutralization test antibodies to BD virus were present in 27 of the in rabbit serum samples. Neutralization test was sensitive in comparison with KFA and CELISA. In comparison with IFA, CELISA was both sensitive and specific in detecting BD viral antibodies. These results extend observations made with laboratory animals to the diagnosis of naturally infected animals.

Evaluation of a cell enzyme-linked immunosorbent assay for the detection of Borna disease virus antibodies in experimentally infected animals

이두식,Lee, Du-sik The Korean Society of Veterinary Science 1992 大韓獸醫學會誌 Vol.32 No.3

보르나병 바이러스 혈청학적 진단법에 있어서 새로 개발된 세포효소면역법의 항체검출에 대한 평가를 위하여 지금까지 주로 사용되어진 간접형광항체법과 네 종류의 실험동물에서 서로 비교하였다. 모든 동물에서 간접형광항체법의 역가와 세포효소면역반응치 사이에 상관계수가 모두 0.8 이상으로 고도의 유의성이 인정되었고 두 진단법의 일치율은 한 희석단계 이하로서 재현성이 아주 좋았다. 세포효소면역반응법은 보르나병 바이러스의 혈청역학적 조사 및 병인기전 연구에 유용하게 이용될 수 있을 것으로 생각된다. The value of the cell enzyme-linked immunosorbent assay as a possible replacement for the indirect immunofluorescence antibody test for the estimation of antibodies against BD virus was assessed in four animal models. The serum antibody response was measured by both assay systems;the variability of both tests was less than one diluent step, and correlation of the two tests was assessed using regression analysis. The study showed that the all four animal models gave satisfactory correlation of CELISA and IFA. There, CELISA is acceptable for use in mouse, rabbit, chicken and rat models.

Detection of human immunodeficiency virus(HIV) type 1 provial sequences of peripheral blood mononuclear cells by using polymerase chain reaction

이두식(Du Sik Lee),임윤규(Yoon Kyu Lim) 한국예방수의학회 1998 예방수의학회지 Vol.22 No.4

중합효소연쇄반응(PCR)을 이용하여 HIV-1에 대하여 혈청반응 양성 및 음성인 동성연애자들의 비배양된 말초혈액 단핵세포로부터 HIV-1 DNA를 검출하는 것이 시도되어졌다. DNA sequence들의 증폭을 위하여 HIV-1 유전자들중 gag유전자 부위에 위치하는 서로 상보적인 primer쌍인 SK38/39가 사용되어졌고 SK9를 probe로써 이용하였다. 혈청반응 양성인 동성연애자 6명중 3명의 말초혈액 단핵세포에서 HIV-1 sequence를 검출할 수 있었으며, 역시 혈청반응 음성인 동성연애자 6명중 3명의 말초혈액 단핵세포에서 HIV-1 sequence를 증명할 수 있었다. 그러나 혈청반응 음성이면서 정상적인 성관계를 하는 대조군에서는 HIV-1 sequence를 검출할 수 없었다. PCR법은 혈청반응 양성인 환자에서는 혈청반응에 보충하여, 그리고 HIV-1에 감염하였지만 혈청반응이 음성인 환자에 있어서는 확진을 내리는데 이용될 수 있을 것으로 사료된다.

증례보고 : 흑기러기에 있어서 Aspergillus fumigatus 에 의한 감염증

신태균,이헌준,이두식,권오덕,양기천,김옥녀,김우택 (Tae Kyun Shin,Hun Jun Lee,Du Sik Lee,Oh Deog Kwon,Ki Chun Yang,Ok Nyu Kim and Woo Taek Kim) 한국임상수의학회 1996 한국임상수의학회지 Vol.13 No.2

국내 말로부터 비폐렴바이러스의 분리 및 면역원성에 관한 연구 I. 말 비폐렴바이러스(equine rhinopneumonitis virus)에 대한 혈청학적 역학조사

조길재,김봉환,이두식,신태균,양기천,임윤규,조성수,Cho, Gil-jae,Kim, Bong-hwan,Lee, Du-sik,Shin, Tae-kyun,Yang, Ki-chun,Lim, Yoon-kyu,Cho, Sung-soo 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.4

The present study was conducted to investigate the incidence of equine herpesvirus 1(EHV-1) infection in Korean horses by cell EUSA and serum neutralization test A significant level of antibodies to EHV-1 was detected from 539(82.8%) by cell EUSA and from 524 horses(80.5%) by SNT indicating that EHV-1 infection in Korean horses are widely spreaded throughout the country.

국내 말로부터 비폐렴바이러스의 분리 및 면역원성에 관한 연구 II. 국내 분리 말 비폐렴바이러스의 특성 및 면역원성 조사

조길재,김봉환,이두식,오문유,고미희,Cho, Gil-jae,Kim, Bong-hwan,Lee, Du-sik,Oh, Moon-you,Ko, Mi-hee 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.4

The study was carried out to characterize the properties of Korean isolates of EHV from aborted fetuses and determine envelope protein profiles. The results obtained were summarized as follows; 1. Two strains of EHV was isolated from 2 liver samples among 10 aborted fetuses from which the virus isolation was attempted. 2. Morphological and some enzymatic properties of the Korean isolates of EHV which was designated as $LC_1$ and $LC_2$ was identical to those of a reference strain of Australia-N of EHV-1. The Korean isolates of EHV could be propagated on ED cell culture and they formed typical plaques 1 to 2 days after infection in the ED cells from which typical cuboidal particles of 150~170 nm diameter herpesvirus were observed. The virus could be detected specifically from neucleus and cytoplasm of infected cells by flourescent antibody technique using FITC labelled anti-Aust IV(EHV-1) antiserum. The Korean isolates, $LC_1$ and $LC_2$ were specifically neutralized by anti Aust IV antiserum and reacted positively to CELISA. 3. The structural polypeptides of purified enveloped virions of $LC_1$ and $LC_2$ isolates of EHV were determined by SDS-polyacrylamide gel electrophoresis to identify the envelope glycoproteins. $LC_1$ and $LC_2$ strains revealed 14 glycoproteins ranging in molecular weight from 190 kD to 31 kD while 17 structural proteins of Aust IV(EHV-1), of which 14 were identical to those of $LC_1$ and $LC_2$, were identified. Upon immunoblotting by rabbit antiserum against EHV isolates and EHV-1(Aust IV), 4 immunogenic proteins of $LC_1$ and $LC_2$ were 135 kD, 88 kD, 64 kD and 59 kD, of which 135 kD, 88 kD and 64 kD proteins were also found in Aust IV(EHV-1).

Demyelination in natural canine distemper encephalomyelitis : An immunohistochemical study of myelin basic protein, myelin associated glycoprotein and glial fibrillary acidic protein in the lesion of demyelination

신태균,권오덕,이두식,이차수,Shin, Tae-kyun,Kwon, Oh-deog,Lee, Du-sik,Lee, Cha-soo The Korean Society of Veterinary Science 1993 大韓獸醫學會誌 Vol.33 No.2

홍역이환개의 중추신경계에서 발생한 수초탈락을 수반하는 뇌척수염을 병리조직학적으로 관찰하고 조직반응과 수축탈락의 과정을 알아보기 위하여 중추신경계 조직을 파라핀 포매, 절편을 만들고 병소가 빈발하는 소뇌와 시신경로는 수초의 주요 구성단백인 MBP와 MAG 그리고 별아교세포의 Marker인 GFAP의 항혈청을 반응시켜 면역세포학적으로 관찰하였다. 조직학적으로는 대뇌에서 신경세포의 일부괴사, 신경아교세포의 결절형성, 소뇌에서는 4뇌실에 인접하여 백질부에서 수초의 공포변성, 수초탈락과 염증세포의 침윤이 현저하였고 피질부의 과립층에 인접한 부위와 시신경로에서도 수초탈락이 인정되었다. MBP와 MAG를 면역반응시킨 결과 수초의 공포화와 수초탈락이 현저한 부위에서는 MBP와 MAG가 동시에 소실되었고 부위에 따라서는 MBP또는 MAG가 먼저 소실되기도 하였다. 동시에 별아교세포의 반응은 수초탈락 초기에는 GFAP양성의 섬유와 세포가 크게 증가한 반면 수초탈락이 심하게 진행된 부위에서는 오히려 소실되는 경향이었다. 따라서 홍역이환 개에서 발생되는 수초탈락은 일차적으로 수초가 공격을 받아 파괴됨을 알 수 있었고 동시에 부위에 따라 다른 소견을 요인별로 비교하였다. Central nervous system of two dogs with natural canine distemper was investigated histopathologically and immunocytochemically with antisera to MBP, MAG and GFAP. Histopathologically, there were neuronal degeneration and diffuse gliosis in the cerebrum, vacuolar degeneration, hypertrophy of astrocytes and demyelination in cerebellar white matter adjacent to the 4th ventricle and optic tracts showing non-inflammatory demyelinating encephalomyelitis (Summers and Appel, 1987). Immunohistochemically, there was a concurrent disappearance of MBP and MAG in the well developed demyelinating lesion in the cerebellar white matter. At the margin of demyelination, Loss of both MBP and MAG varied on the stage of demyelinating process. GFAP-positive astrocytes were hypertrophied and contained canine distemper virus intranuclear inclusions. GFAP-positive fibers were increased at the early stage of demyelination, and then were not immunoreaeted at the well developed demyelination. Hypertrophic astrocytes with intranuclear inclusions were commonly identified in the interfascular layer without myelin vacuolation and demyelination. This is the first study of primary demyelination and astroglial reactions in natural CDE investigated using immunocytochemistry of two myelin proteins and GFAP. Concurrent loss of MBP and MAG suggest that the myelin sheath is the target in the demyelinating process in CDE.

제주마의 혈액형에 관한 연구 II. 혈액 단백질형

조길재,김봉환,이두식,이경갑,Cho, Gil-jae,Kim, Bong-hwan,Lee, Du-sik,Lee, Kyoung-kap 대한수의학회 2000 大韓獸醫學會誌 Vol.40 No.2

The present study was carried out to investigate the blood markers of Cheju horses. The blood protein types (biochemical polymorphism) were tested from 73 Cheju native horses (CNH) and 118 Cheju racehorses(CRH) by horizontal polyacrylamide gel electrophoresis (HPAGE), isoelectric focusing (IEF) and starch gel electrophoresis (SGE). At the same time, their phenotypes and gene frequencies were studied. The biochemical polymorphism phenotypes observed with high frequency were A1B-KK(97.3%), ALB-AB(49.3%), AP-SS(100%), ES-II(30.1%), GC-FF(87.7%), HB-BIBI(49.3%), TF-F2R(41.1%), TF-EF2(8.2%), PGD-FF(97.3%), PGM-SS(50.7%), GPI-II(74.0%) in CNH, While A1B-KK(99.2%), ALB-BB(50.8%), AP-SS(99.2%), ES-II(42.4%), ES-IS(14.4%), GC-FF(95.8%), HBB-IB II(39.8%), TF-F2R(21.2%), PGD-FF(77.1%), PGD-SS(4.3%), PGM-SS(72.9%), GPI-II(90.7%) in CRH. Alleles observed with high frequency were $AlB^{K}$(0.986), $ALB^{B}$(0.616), $AP^{S}$(1.000), $ES^{I}$(0.479), $ES^{F}$(0.274), $GC^{F}$(0.938), $GPI^{I}$(0.856), $HB^{BI}$(0.685), $PGD^{F}$(0.993), $PGM^{S}$(0.753), $TF^{F2}$(0.404), $TF^{R}$(0.397) in CNH and $AlB^{K}$(0.996), $ALB^{B}$(0.720), $AP^{S}$(0.996), $ES^{I}$(0.661), $ES^{F}$(0.203), $GC^{F}$(0.979), $GPI^{I}$(0.936), $HB^{BI}$(0.534), $PGD^{F}$(0.864), $PGM^{S}$(0.852), $TF^{F2}$(0.428), $TF^{R}$(0.272) in CRH. $TF^{E}$(0.041) allele and silent gene($ES^{I{^*}}$ : 0.014) were observed in CNH. The mean heterozygosity in CNH and CRH was observed 0.2974 and 0.2864, respectively.

SDS 처리한 브루셀라 항원과 Yersinia enterocolitica 0:9주의 혈청학적 교차반응 연구

임윤규,양기천,이경갑,박전홍,이두식,박용호,강승원,목지원,이영순,Lim, Yoon-kyu,Yang, Ki-chun,Lee, Kyung-kap,Park, Jun-hong,Lee, Du-sik,Park, Yong-ho,Kang, Seung-won,Mok, Ji-won,Lee, Yong-soon 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.1

Brucella abortus cell wall antigen was extracted from Brucella abortus 1119-3 by ultrasonication and followed by sodium dodecyl sulfate(SDS) treatment. In order to confirm whether this preparation is serologically cross reactive with Yersinia enterocolitica 0:9, Western blot analysis with mouse anit-Brucella abortus1119-3 and with mouse anti-Yersinia enterocolitica 0:9 was performed. ELISA results from using those Brucella antigen and Yersinia antigen were assessed whether they had correlation. According to the results of western blot analysis and ELISA, there was no evidence of cross reactivity between the Brucella abortus 1119-3 antigen preparation and Yersinia enterocolitica 0:9. Therefore the SDS treated antigen prepared in this study could be suitably used as specific ELISA antigen without confusion in the interpretation of serological tests for brucellosis in cattle.

국내 말로부터 비폐렴바이러스의 분리 및 면역원성에 관한 연구 III. 말 비폐렴바이러스의 실험동물에 대한 면역원성 조사

조길재,김봉환,소신희,이두식,Cho, Gil-jae,Kim, Bong-hwan,So, Sin-hee,Lee, Du-sik 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.4

The study was carried out to investigate the pathogenicity of EHV isolate to hamsters and mice and immunogenicity of experimentally produced. vaccine were evaluated in the horses. Hamsters infected. intranasally with $LC_1$ isolate showed symptoms of nasal discharge, conjunctivitis and body weight loss during the observation period of 12 days after infection, while only slight depression and body weight loss were noticed with mice infected with $LC_1$ indicating that hamsters are more susceptible to the virus. Antibody titer of mice and hamsters were gradually increased to highest level of 1:2560~10240, 1:640~1280, respectively, at 7~12 days post vaccination. Horses immunized against $LC_1$ killed vaccine reached to maximum antibody titer of 1:20480 around 4 weeks after 1st vaccination and declined after 12 weeks post vaccination. No significant antibody increase were detected after 2nd vaccination. Mean body temperature and mean total leukocyte counts remained within normal range and no adverse reaction were noticed after vaccination.