• 대한수의학회지
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• 제39권6호
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• pp.1119-1125
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• 1999

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.

• 한국동물위생학회지
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• 제32권2호
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• pp.125-129
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• 2009

SBovine tuberculosis is a chronic bacterial disease of animals and humans caused by Mycobacterium bovis. Besides the classical intradermal tuberculin test, a number of blood and serum tests have been used. The purpose of this study was to establish seroprevalence of M. bovis. The sera were screened using the ELISA technique. A total seroprevalence of 65.8% in positive cattle, suspect 36.0%, negative 5.9% in TB-infected herds by PPD and dairy cattle is 3.0%, Hanwoo is 1.6% in TB-free herds. The deer of seroprevalence is 55.0% in TB-infected herd and 7.7% in TB-free herds.

• 대한미생물학회지
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• 제6권1호
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• pp.41-60
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• 1971

The author prepared a tuberculin sample using synthetic medium for the test material in this experiment. For the test animals, guinea pigs, healthy calf sensitized with heat killed tubercle bacilli, naturally infected dairy cattle, goat, and calf were used. A reduced reactivity to tuberculin was observed when the second tuberculin test was done within 28 days after the first. The results obtained are summarized as follows. 1. The experimental HCSM tuberculin was shown the similar activities as the control products of standard tuberculin. 2. The potency of the experimental tuberculin preserved at $5^{\circ}C$ in refrigerator maintained the activity for three years. 3. By the intradermal test, the experimental tuberculin was proved to be specific to detect tuberculous dairy cattles and goats. 4. The appearance of skin reaction to tuberculin in naturally infected dairy cattles, goat and beef cattles was soft and diffuse swellings or circumscribed swellings accompanied by induration. 5. In the case of observing tuberculous dairy cattles, goats and beef cattles, as the criterion of the reaction to the caudal fold test, results were taken as positive when findings are either hard and circumscribed or soft and diffuse swellings of 3 mm at least in size. 6. The acid fast organisms isolated from tuberculin positive animals were identified mostly as M. tuberculosis var. bovis. 7. When the tuberculin test was repeated in the tuberculin positive animals, the reactivity decreased rapidly, lasting 4 weeks at least.

• Pediatric Infection and Vaccine
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• 제9권2호
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• pp.208-214
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• 2002

목 적 : BCG 접종 후 투베르쿨린 반응은 사용한 백신의 종류, 백신의 상태, 투여방법, 접종시 연령, 접종후 검사하기까지의 시간 등의 다양한 요인에 따라 다르게 나타난다. 최근 흉터가 작고 이상반응이 적은 경피용 BCG의 접종이 증가하고 있으나 투베르쿨린 반응 검사 및 예방 효과 등에 대한 자료가 거의 없는 실정이다. 본 연구는 개원가에서 실시한 BCG 및 투베르쿨린 반응을 평가하고 피내 BCG 백신과 경피용 BCG 백신과의 비교, 5TU PPD와 2TU PPD의 비교를 실시하였다. 방 법 : 1997년 2월부터 2001년 6월까지 세군데의 개인 소아과 의원에서 생후 1개월경 BCG 접종을 받은 후 투베르쿨린 반응을 실시한 200명의 소아를 대상으로 의무기록지를 이용한 후향성 조사를 실시하였다. 이 중 151명은 Tokyo 172주로 경피 접종하였고 49명은 피내 접종(28명:copenhagen주, 1명:French 1173P2주, 20명:미상)하였다. 경피 접종한 151명 중 129명(64.5%)은 2TU, 22명(11%)은 5TU PPD를 사용하였고 피내 접종한 49명 중 35명(17.5%)은 2TU, 14명(7%)은 5TU PPD를 사용하였다. 결 과 : 경피용 BCG 접종군에서 2TU PPD와 5TU PPD에 대한 경결의 크기(양성률)는 $9.2{\pm}4.4mm$(86.8%), $12.4{\pm}3.5mm$(95.5%)로 5TU PPD로 검사한 경우가 2TU PPD로 검사한 경우보다 통계적으로 유의하게 더 크게 나타났다(P=0.02). 피내용 BCG 접종군에서 2TU PPD와 5TU PPD에 대한 경결의 크기(양성률)는 각각 $5.7{\pm}5.1mm$(60%), $6.6{\pm}4.8mm$(71.4%)로 두 값 사이에 통계적으로 유의한 차이가 없었다(P=0.569). 2TU PPD에 대한 반응은 경피용 BCG 접종군에서의 경결 크기(양성률)가 $9.2{\pm}4.4mm$(86.8%), 피내용 BCG 접종군에서는 $5.7{\pm}5.1mm$(60%)로 경피용 BCG 접종군에서의 경결크기가 피내용 BCG 접종군에 비해 통계적으로 유의하게 크게 나타났다(P<0.001). 5TU PPD에 대한 반응은 경피용 BCG 접종군에서의 경결크기(양성률)는 $12.4{\pm}3.5mm$(95.5%), 피내용 BCG 접종군에서는 $6.6{\pm}4.8mm$(71.4%)로 역시 경피용 BCG 접종군에서 경결크기가 피내용 BCG 접종군에 비해 통계적으로 유의하게 크게 나타났다(P<0.001). 결 론 : 경피용 BCG(Tokyo 172균주) 접종군의 2TU PPD와 5TU PPD에 대한 반응은 피내용 BCG 접종군에 비해 경결의 크기가 유의하게 컸고, 경피용 BCG 접종군에서 5TU PPD의 경결 크기가 2TU PPD에 비해 유의하게 크게 나타났으므로 경피용 BCG 환아들에게서의 결핵 반응 검사 결과 판독의 새로운 기준이 수립되어야 할 것으로 사료된다.

• 한국동물위생학회지
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• 제41권2호
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• pp.71-78
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• 2018

Bovine tuberculosis (bTB) is a wide-spread zoonotic disease in cattle, which is casued by Mycobacterium bovis that is a part of Mycobacterium tuberculosis complex (MTC). This study describes a field trial conducted in 42 herds with the history of prevalence bovine tuberculosis. Two cell-mediated immunity tests, the gamma-interferon (${\gamma}-IFN$) assay and the single intradermal tuberculin test (SIT) were applied for the diagnosis of bovine tuberculosis in 5,289 animals. The ${\gamma}-IFN$ assay presented 144 (2.7%) head of cattle with the positive result, and 112 (2.1%) head of cattle were shown to be bTB-positive by the SIT. The positive concordance was 45.5%, and the negative concordance was 98.2%. The ${\gamma}-IFN$ assay showed more positive results in younger cattle, especially between 12 and 23 months of age. It is shown that the strategic combination of both cell-mediated immunity test methods is more efficient for the detection of bTB to reduce the number of false positive individuals which are being slaughtered.

• 대한미생물학회지
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• 제5권1호
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• pp.41-48
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• 1970

In order to assay one lot of the potency of tuberculin for intradermic use, four guinea pigs sensitized with heat killed M. tuberculosis were used by the method originally of the Agriculture Research Service Center(A. R. S.) and Bureau of Animal. Industry(B. A. I.) in the United. States of America in which guinea pigs were used on 21st day after sensitization. There is doubt whether the guinea pigs are sensitized or not. Therefore, this study has taken the method of Middle Brook-Dubos hemagglutination test, and observed the humoral hemagglutinating antibody in the blood. The results obtained are as follows. 1. The maximum titer of hemagglutinating antibody was demonstrated 30days after sensitizing the guinea pigs with killed tubercle bacilli. The maximum titer was continued for a period of 45 days and thereafter declined gradually. 2. The swelling and redness of the region of the sensitized guinea pigs was shown to be reached to the maximum size within 24 hours after intradermal inoculation with HCSM tuberculin and gradually reduced 96 hours after inoculation. The swelling and redness could not be detected by macro-observation after 14 days.

• 대한수의학회지
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• 제41권4호
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• pp.535-542
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• 2001

A multiplex PCR technique was developed for detecting specifically each Mycobacterium bovis, M. tuberculosis, M. avium and M. avium subsp, paratuberculosis, respectively, using clinical samples of field cattle. To apply this novel technique to clinical specimens, blood sample was obtained from live cows comprising 11 intradermal tuberculin test (ITT)-positive and 17 ITT-negative and tested by multiplex PCR. Positive results were obtained from 15 cows by the multiplex PCR, showing that 4 (23.5%) of the 17 ITT-negative cows were multiplex PCR positive. The multiplex PCR results also showed that among the 15 positive cows, 7 (46.7%) were infected with M. bovis, 1 (6.7%) with M. tuberculosis and 7 (46.7%) with M. avium. The sensitivity and specificity of multiplex PCR in comparison with those of ITT were 100% and 76.5%. The correlation between the multiplex PCR and ITT assays with blood samples was considered excellent, 85.7% agreement and ${\kappa}=0.72$. The results obtained, using reference mycobacterial strains and typed clinical samples, show that the multiplex PCR method may be a rapid, sensitive, and specific tool for the differential identification of various mycobacterial strains in a single-step assay. Therefore, multiplex PCR assay is a useful tool for early diagnosis of tuberculosis in live cattle and to identify the species or complex of mycobacterium from clinical samples.

• 한국동물위생학회지
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• 제33권4호
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• pp.335-340
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• 2010

On the basis of the 2009 business plan, 20,394 Korean native cattle and beef cattle were carried examination of bovine tuberculosis by using ELISA technique from March to December. As a result, 66 cattle tested positive for tuberculosis and showed 0.32% positive ratio. Intradermal tuberculin test about 66 cases of ELISA positive cattle was carried out, and all of 66 cattle were confirmed as negative. However, when 7 PPD-positive cattle derived from slaughterhouse were tested by 20k ELISA kit and MS ELISA kit, 3 (2 suspect) cattle and 5 cattle showed positive results, respectively. As compared to the results of PPD test, the concordance rates were 43% (71% included suspect) with 20k ELISA kit and 71% with MS ELISA kit.

• Pediatric Infection and Vaccine
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• 제8권2호
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• pp.175-180
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• 2001

목 적 : 최근 국내에서 1998년부터 5TU 대신 2TU를 사용함으로써 결과 판정에 어려움이 있다. 5TU와 2TU에 대한 투베르쿨린 반응 정도는 차이가 있게 마련이나 아직까지 적절한 국내 기준이 마련되어 있지 않은 실정이기에 생후 4주 이내에 경피 다자법으로 BCG를 접종 받은 영아에서의 PPD 1TU, 2TU 및 5TU에 대한 반응을 관찰하고자 하였다. 방 법 : 2000년 1월부터 2000년 8월까지 8개월간 순천향대학병원 육아지도회에서 결핵의 가족력, 피부 질환, 영양 장애 및 면역 기능 저하 소견이 없는 건강한 신생아 중 생후 4주 이내에 경피 다자법(Tokyo strain, Japan BCG Laboratory Co.)으로 BCG를 접종한 92명(남아 : 56명, 여아 : 36명)에게 생후 4개월에 PPD 1TU, 2TU 및 5TU로 결핵 피부반응 검사를 시행한 후 그 반응을 관찰하였다. 통계처리는 통계 프로그램인 SPSS 10.0을 이용하여 ANOVA, Chi-square test을 적용하여 P<0.05인 경우 통계적으로 유의하다고 판단하였다. 결 과 : 1) 전체 92명 중 31명에서 1TU, 31명에서 2TU 그리고 30명에서는 5TU를 사용하여 결핵 피부반응 검사를 시행하였다. 1TU로 검사시 경결의 크기가 0 mm인 경우가 가장 많았고 그 다음으로 7 mm, 6 mm 순으로 많았다. 2TU로 검사시 경우 8 mm가 가장 많았으며 그 다음은 3 mm, 6 mm, 10 mm가 많았다. 5TU로 검사시 13 mm와 6 mm가 가장 많았다. 2) 경결의 평균 크기는 PPD 1TU, 2TU 및 5TU로 검사하였을 때 각각 $5.7{\pm}4.2mm$, $7.1{\pm}3.7mm$, $9.2{\pm}4.2mm$로 1TU와 2TU는 통계학적으로 차이가 없었으나 5TU의 경우 PPD 1, 2TU에 비해 평균 경결 크기가 의미 있게 컸다(P<0.05). 3) 경결의 크기가 10 mm 이상인 경우는 1TU에서는 31명 중 6명(19.4%), 2TU에서는 31명 중 9명(29%), 5TU에서는 30명 중 16명(53.3%)이었으며 1TU와 2TU는 통계학적으로 차이가 없었으나 5TU의 경우 PPD 1, 2TU에 비해 의미 있게 많았다(P<0.05). 경결의 크기가 5 mm 이상인 경우는 통계학적 유의성은 없었지만 5TU로 검사시 가장 많았다. BCG 반흔 수는 1TU의 경우 $13.81{\pm}4.48$개, 2TU는 $13.06{\pm}4.18$개, 5TU는 $13.07{\pm}3.07$개로 차이가 없었다. 결 론 : 현재 국내에서 사용되고 있는 2TU을 사용한 결핵 피부반응 검사에 대한 새로운 평가 기준이 필요하고 결핵의 진단기준에 대한 더 많은 연구가 필요하다고 사료된다.

• 한국동물위생학회지
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• 제34권4호
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• pp.333-339
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• 2011

Loop-mediated isothermal amplification (LAMP) was developed to detect Mycobacterium tuberculosis complex (MTC) and non-tuberculous mycobacterium (NTM) genomic DNA in blood samples of Korea native cattle. A set of four primers, two outer and two inner, were designed from M. bovis and M. avium genomic DNA targeting the IS6110 and 16S rRNA gene, respectively. Based on 85 Intradermal Tuberculin Test (ITT) positive blood sample and using conventional PCR and LAMP, the agreement quotient (kappa), which measures agreement beyond chance were 0.93 (conventional PCR) and 0.97 (LAMP), respectively. The detection limit of the LAMP method was $2.0{\times}10^2$ copy/ml M. bovis and M. avium cells, compared to $2.0{\times}10^3$ copy/ml M. bovis and M. avium cells for conventional PCR. These results suggest that the LAMP is a powerful tool for rapid, sensitive, and practical detection of MTC and NTM in blood samples of Korea native cattle.