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비교 피내 검사와 감마 인터페론 검사에 의한 우결핵의 진단
조윤상,김종만,정석찬,우승룡,김종염,유한상,박용호,안종삼,Cho, Yun-sang,Kim, Jong-man,Jung, Suk-chan,Woo, Seung-ryong,Kim, Jong-yeom,Yoo, Han-sang,Park, Yong-ho,Ahn, Jong-sam 대한수의학회 1999 大韓獸醫學會誌 Vol.39 No.6
Since Robert Koch found tubercle bacilli in 1882, the studies on tubercle bacilli of human and animal had been carried out. Being old tuberculin(OT) introduced in 1890, the specificity of the diagnosis of tuberculosis has been improved by continual uses of heat concentrated synthetic medium(HCSM) and purified protein derivatives(PPD) tuberculin. Now, two types of tuberculin test are used worldwidly ; the single intradermal test(SIT) using bovine tuberculin and the single intradermal comparative tuberculin test(SICTT) using avian and bovine tuberculins. In the SICTT, each countries have used with different combination of both avian and bovine tuberculins' titers. However, this kinds of studies have not reported in Korea. Therefore, the studies on the combination of their tuberculins' titers were performed through intradermal test of guinea pigs sensitized with either Mycobacterium bovis or M avium and were examined in 10 cattles of SIT positive reactors. Also, IFN-${\gamma}$ assay, the latest diagnostic method of bovine tuberculosis, was experimentally applied to SIT positive reactors. For determining the optimal titers, sensitized guinea pigs with M bovis and M avium were intradermally injected avian and bovine tuberculin. In guinea pigs sensitized with M bovis, bovine tuberculin 50 T.U. showed significant difference from all tested concentrations of avian tuberculin(p < 0.05). In guinea pigs sensitized with M avium, there is significantly different between bovine tuberculin and avian tuberculin by 25 T.U.(p < 0.01). Therefore, optimal titers of bovine and avian PPD tuberculins' titers for the SICTT in Korea were 5,000 and 2,500 tuberculin units, respectively, and the swelling diffences between bovine and avian site in SIT positive reactors were above 3mm. Also, in IFN-${\gamma}$ assay, the 9 SIT positive reactors were showed all the positive reactions.
장구균의 vancomycin 내성 유전자와 종 특이유전자의 검출을 위한 Multiplex polymerase chain reaction 개발
조윤상,이희수,김종만,안종삼,류판동,박용호,유한상,이문한,Cho, Yun-Sang,Lee, Hee-Soo,Kim, Jong-Man,Ahn, Jong-Sam,Ryu, Pan-Dong,Park, Yong-Ho,Yoo, Han-Sang,Lee, Mun-Han 대한수의학회 2003 大韓獸醫學會誌 Vol.43 No.1
A multiplex PCR assay, which allows simultaneous detection of vancomycin resistant genotypes and Enterococcus species-specific genes, was developed. Vancomycin resistant enterococci (VRE) from chickens and humans could be detected for vanA, vanB, vanC-1, vanC-2, $ddl_{E.faecium}$ and $ddl_{E.faecalis}$ by multiplex PCR. Eight isolates of VRE from humans (n=11) had $ddl_{E.faecium}$ and vanA, and 3 isolates of the VRE had $ddl_{E.faecium}$ and vanB. One isolate of VRE from chickens (n=6) had $ddl_{E.faecium}$ and vanA, and 5 isolates of the VRE had only vanA. E. faecium, E. faecalis, E. gallinarum and E. casseliflavus were also confirmed for the species-specific gene by multiplex PCR. This multiplex PCR could detect E. faecium, E. faecalis, E. gallinarum, E. casseliflavus, vanA, vanB, vanC-1 and vanC-2, simultaneously. The PCR assay established in the present study can be an alternative to time-consuming biochemical tests and antibiotic susceptibility tests of Enterococcus spp.
살모넬라 세포외막단백질 혼합백신을 이용한 돼지 및 닭에서의 살모넬라균감염증 방어효과
이희수,임숙경,조윤상,주이석,김재학,김종만,Lee, Hee-Soo,Lim, Suk-Kyung,Cho, Yun-Sang,Joo, Yi-Seok,Kim, Jae-Hak,Kim, Jong-Man 대한수의학회 2007 大韓獸醫學會誌 Vol.47 No.2
We investigated the safety, immunogenicity and protectivity of mix-crude outer membrane protein (cOMP) vaccine against salmonellosis in animals. The mix-cOMP vaccine was extracted from Salmonella enterica serovar Typhimurium (ST) and Salmonella enterica serovar Enteritidis (SE) and Salmonella enterica serovar Braenderup (SB) isolated from pigs. The mix-cOMP vaccine gave significantly higher antibody response than ST-bacterin and ST-cOMP vaccine in guinea pigs. The survival rates of mix-cOMP vaccinated groups showed significantly higher (100%) than those (0-20%) of unvaccinated control group, challenged with 3 species of Salmonella (ST, SE and SB) in mice. Vaccinated groups in pigs showed reduction of clinical signs, increase of average weight gains, decrease of bacterial recovery rates, compared with unvaccinated groups. Especially, the survival rates (100%) of vaccinated groups in chickens showed higher than that (0%) of unvaccinated group. Based on these results, we suggest that the mix-cOMP Salmonella vaccine developed in this study will be effective for the protection against Salmonellosis caused by the various serotypes Salmonella species in animals.
Development of a multiplex-PCR for the rapid detection of Escherichia coli O157:H7 from raw beef
정석찬,정병열,윤장원,조윤상,김종염,박용호,Jung, Suk-chan,Jung, Byeong-yeal,Yoon, Jang-won,Cho, Yun-sang,Kim, Jong-yeom,Park, Yong-ho The Korean Society of Veterinary Science 1998 大韓獸醫學會誌 Vol.38 No.1
Esherichia coli O157 : H7의 slt I, slt II, uid A 및 eaeA 4종 유전자를 동시에 검출하기 위한 multiplex PCR 기법을 확립하고 쇠고기중 직접 E coli O157 : H7 검출시험을 실시하였다. 4 set의 primers를 이용한 multiplex PCR 기법으로 31종의 장내세균에 대한 특이성을 조사한 결과 E coli O157 : H7 에서 1,087bp (eae A), 584bp (slt II), 348bp (slt I) 또는 252bp (uid A)크기의 DNA를 동시에 특이적으로 검출할 수 있었다. E coli O157 : H7 15주는 모두 uid A 및 eae A 유전자가 동시에 검출되었고, 다른 장내세균에서는 검출되지 않았다. slt I 또는 slt II 유전자를 가지고 있는 E coli 표준균주 24종을 이용하여 multiplex PCR 기법과 Vero cell cytotoxicity assay을 비교검사한 결과 베로톡신 산생능과 PCR법의 결과는 일치하였다. mutiplex PCR 기법의 쇠고기중 검출한계는 modified EC(mEC)에서 증균없이는 E coli O157 : H7균 $10^4cells/g$ 이상에서 검출이 가능하였으나 mEC에 1차 증균후 modified TSB 증균하였을 경우에는 10cells/g이하까지도 검출이 가능하였다. 개발된 multiplex PCR 기법을 쇠고기 40종에 직접 적용한 결과 E coli O157 : H7은 검출되지 않았으나 slt I 및 slt II유전자를 가지고 있는 E coli 4종이 검출되었으며, 이들의 혈청형은 O6, O112, O115 및 O139 였다. 이 연구에서 개발된 multiplex PCR은 쇠고기중 E coli O157 : H7을 신속하고 특이적으로 검출하는데 사용할 수 있을 것으로 사료된다.
난황면역제를 이용한 개 주요 소화기 및 호흡기질병 방제에 관한 연구 I. 개 보데텔라, 파보바이러스 및 개디스템퍼 항원의 닭에서의 면역반응
이희수,김종만,우승룡,정병열,조윤상,탁동섭,임숙경,유한상,윤용덕,허원,문영식,오진식,Lee, Hee-soo,Kim, Jong-man,Woo, Seng-ryong,Jung, Byeong-yeal,Cho, Yun-Sang,Tark, Dong-seob,Lim, Sook-kyoung,Yoo, Han sang,Yoon, Yong-dhuk,Huh, Woo,Mun, Y 대한수의학회 2004 大韓獸醫學會誌 Vol.44 No.1
This study was carried out to produce IgY against B. bronchiseptica, parvovirus and distemper virus that are major pathogens in alimentary and/or respiratory diseases of dogs. In the comparison of adjuvants, ISA70 was the best in the rapid induction and maintence of antibody titers. Agglutination antibody titers against B. bronchiseptica were 1:1,280 ~ 1:10,240 in sera and 1:160 ~ 1:1,280 in egg yolk. Hemagglutination inhibition(HI) titers against parvovirus in sera and egg yolk were 1:80 ~ 1:320 and 1:64 ~ 1:256, respectively. Virus neutralization titers against canine distemper was 1:8 ~ 1:64 in sera and egg yolk. These results suggested that egg yolk antibody titers could be variable according to a sort of adjuvant and antigens of the pathogens.
난황면역제를 이용한 개 주요 소화기 및 호흡기질병 방제에 관한 연구 II. 난황면역제의 실험동물 및 개에 있어서의 질병방제 효과
이희수,김종만,우승룡,정병열,조윤상,유한상,윤용덕,허원,문영식,오진식,Lee, Hee-Soo,Kim, Jong-man,Woo, Seung-ryong,Jeong, Byeong-yeal,Cho, Yun-Sang,Yoo, Han-sang,Yoon, Yong-dhuk,Huh, Won,Mun, Young-sik,Oh, Jin-sik 대한수의학회 2004 大韓獸醫學會誌 Vol.44 No.3
Immunoprophylatic effect of IgY against B. bronchispetica was proven with 100% preventive rate in mice administrated with IgY with antibody titer 1:640~1:2,560. Intramuscular administration was more efficient than oral administration. This phenomenon was also observed in the therapeutic effects of IgY after challenge with B. bronchseptica in mice. In the field trials with the egg yolk antibodies from hens immunized with combined antigens with B. bronchiseptica and parvovirus, curing rates in dogs with severe clnical signs such as bloody diarrhea were 81.6% and 86.7% by intramuscular or subcutaneous administration of IgY, respectively. Safety of the antibodies in dogs was proven without any side effects such as vomiting, edema, fever, etc. by adminstration of double doses for 7 days. These results indicated that the egg yolk antibodies could be used as effective prevention and treatment of alimentary and respiratory diseases in dogs.
국내 우군에서 소 결핵 진단을 위한 피내검사법과 Interferon-γ 생성 검사의 비교
신승원,신민경,차승빈,우종태,이성모,구복경,조윤상,정석찬,유한상,Shin, Seung Won,Shin, Min Kyoung,Cha, Seung Bin,Woo, Jong Tae,Lee, Sung Mo,Ku, Bok Kyung,Cho, Yun Sang,Jung, Suk Chan,Yoo, Han Sang 대한수의학회 2011 大韓獸醫學會誌 Vol.51 No.2
Bovine tuberculosis (bTB), caused primarily by Mycobacterium bovis, continues to exert an economic loss, even in countries with active control measures, and is one of zoonotic diseases enable to be transmitted to human. The control and eradication of bTB are mainly based on a test and slaughter policy and/or abattoir surveillance. Various factors including limitation of diagnostic tests have been considered as major constraints to eradication. Single intradermal test (SIT) is the official diagnostic test. New diagnostic methods are needed to be developed, because of limitations of the test. In the present study SIT was compared with single intradermal comparative cervical test (SICCT) and interferon (IFN)-${\gamma}$ assay. There was very low correlation between SIT and SICCT. However, high correlation was shown between SIT and IFN-${\gamma}$ assay while no correlation was observed between SICCT and IFN-${\gamma}$ assay. Therefore, our results suggest the possibility of replacement of SIT with IFN-${\gamma}$ assay for the diagnosis of bovine tuberculosis.