• Korean Journal of Veterinary Research
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• v.39 no.6
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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.

• Korean Journal of Veterinary Service
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• v.32 no.2
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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.

• The Journal of the Korean Society for Microbiology
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• v.6 no.1
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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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• v.9 no.2
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• pp.208-214
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• 2002

Purpose : The number of newborns vaccinated with BCG of Tokyo 172 strain, which has been claimed to cause lesser degree of local adverse events including scar, has recently been increasing. However, tuberculin response to this vaccine has inadequately been studied, especially with newborns cared at primary care clinics. We, therefore, performed a study in newborns vaccinated with BCG at private pediatric offices and evaluated the response to PPD 2TU or PPD 5TU following vaccination with percutaneous or intradermal BCG. Methods : Two hundred infants who had been cared at three private pediatric offices were retrospectively enrolled in the study. One hundred fifty one infants had received percutaneous BCG(Tokyo strain); 129 infants had had tuberuclin test with PPD 2TU and the rest of 22 infants with PPD 5TU. Forty nine infants had received intradermal BCG(28 infants Copenhagen strain, I infant French strain, 20 infants unknown); 35 infants had had tuberculin test with PPD 2TU, 14 infants(11%) with PPD 5TU. Results : In infants vaccinated with percutaneous BCG, the mean induration diameter in tuberculin test was significantly greater with PPD 5TU($12.4{\pm}3.5mm$) compared to PPD 2TU ($9.2{\pm}4.4mm$). In infants vaccinated with intradermal BCG, the mean induration diameters in tuberculin test were $5.7{\pm}5.1mm$ to PPD 2TU and $6.6{\pm}4.8mm$ to PPD 5TU, which were not significantly different. The tuberculin response to PPD 2TU was significantly greater in infants vaccinated with percutaneous BCG compared to those with intradermal BCG. The tuberculin response to PPD 5TU was also significantly greater in infants vaccinated with percutaneous BCG compared to those with intradermal BCG. Conclusion : Percutaneous BCG(Tokyo strain) seems to cause greater response to tuberculin compared to intradermal BCG and PPD 2TU induces weaker response compared to PPD 5TU. Acknowledging some discrepancies from the previously reported data, which might have been due to the different source of the study subjects, more studies are needed to establish the range of tuberculin response following BCG vaccination in order to differentiate from tuberculosis.

• Korean Journal of Veterinary Service
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• v.41 no.2
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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.

• The Journal of the Korean Society for Microbiology
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• v.5 no.1
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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.

• Korean Journal of Veterinary Research
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• v.41 no.4
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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.

• Korean Journal of Veterinary Service
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• v.33 no.4
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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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• v.8 no.2
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• pp.175-180
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• 2001

Purpose : Recently in Korea since 1998, 2TU has been used instead of 5TU, which caused a confusion in interpreting tuberculin skin tests. We compared the tuberculin skin reaction to PPD(purified protein derivatives) 1TU, 2TU and 5TU. Methods : We studied tuberculin skin reaction to PPD 1TU, 2TU and 5TU on 92 infants who were vaccinated with percutaneous multiple puncture BCG vaccine(Tokyo 172 strain, Japan BCG Laboratory Co.) in neonatal period. Diameters of indurations were measured in millimeters 48~72 hours after the intradermal injection of PPD 1TU, 2TU and 5TU. Results : Among 92 infants(male : 56, female : 36), 1TU was used on 31 infants, 2TU on 31 infants and 5TU on 30 infants for tuberculin skin test. The mean diameter of induration for PPD 1TU, 2TU and 5TU were $5.7{\pm}4.2mm$, $7.1{\pm}3.7mm$, and $9.2{\pm}4.2mm$, respectively. There was no statistical difference in mean diameter of induration between 1TU and 2TU. However, when tested with 5TU, the diameter of induration was significantly larger than those of PPD 1TU and 2TU(P<0.05). Six subjects(19.4%) tested with 1TU, 9 subjects(29%) tested with 2TU and 16 subjects(53.3%) tested with 5TU had diameter of 10 mm or more. There was no statistical difference between 1TU and 2TU but for 5TU, number of subjects with diameter of 10 mm or more was significantly higher than PPD 1TU and 2TU(P<0.05). Number of subjects with induration diameter of 5 mm or more for 1TU, 2TU and 5TU was 19(61.3%), 22(71.0%) and 26(86.7%), respectively. However, there was no statistical difference. Conclusion : A new standard criteria for interpreting the tuberculin skin test with PPD 2TU should be established for clinical diagnosis of tuberculosis.

• Korean Journal of Veterinary Service
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• v.34 no.4
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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.