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Development of analytical method for determination of spinetoram residues in livestock using LC-MS/MS (LC-MS/MS를 이용한 축산물 중 Spinetoram 공정시험법 개발 및 검증)

  • Ko, Ah-Young;Kim, Heejung;Do, Jung Ah;Jang, Jin;Lee, Eun Hyang;Ju, Yun Ji;Kim, Ji Young;Chang, Moon-Ik;Rhee, Gyu-Seek
    • Analytical Science and Technology
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    • v.29 no.2
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    • pp.94-103
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    • 2016
  • An analytical method was developed to determine the amount of spinetoram (spinetoram J and spinetoram L) in livestock samples. The spinetoram was extracted with acetonitrile and purified through a primary secondary amine (PSA) sorbent. The spinetoram residues were then quantified and confirmed using a liquid chromatography–tandem mass spectrometer (LC-MS/MS) in the positive ion mode using multiple reactions monitoring (MRM). Matrix-matched calibration curves were linear over the calibration ranges (0.005-0.5 mg/kg) into a blank extract with r2 > 0.994. The limits of detection and quantification were 0.002 and 0.01 mg/kg, respectively. The recovery results of spinetram ranged between 81.9-106.4% at different concentration levels (LOQ, 10LOQ, 50LOQ, n=5) with relative standard deviations (RSDs) less than 10%. All values were consistent with the criteria ranges requested in the Codex guidelines (CAC/GL40, 2003). An interlaboratory study was conducted to validate the method. The proposed analytical method proved to be accurate, effective, and sensitive for spinetoram determination. The method will be used as an official analytical method in Korea.

Evaluation of X-ray System for Nondestructive Testing on Radioactive Waste Drums (방사성폐기물 드럼 비파괴 검사를 위한 X-ray 장비 평가)

  • Park, Jong-Kil;Maeng, Seong-Jun;Lee, Yeon-Ee;Hwang, Tae-Won
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.6 no.3
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    • pp.189-203
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    • 2008
  • The physical and chemical properties of radioactive waste drums, which have been temporarily stored on site, should be characterized before their shipment to a disposal facility in order to prove that the properties meet the acceptance guideline. The investigation of NDT(Nondestructive Test) method was figured out that the contents in drum, the quantitative analysis of free standing water and void fraction can be examined with X-ray NDT techniques. This paper describes the characteristics of X-ray NDT such as its principles, the considerations for selection of X-ray system, etc. And then, the waste drum characteristics such as drum type and dimension, contents in drum, etc. were examined, which are necessary to estimate the optimal X-ray energy for NDT of a drum. The estimation results were that: $(R)\acute{A}$ the proper X-ray energy is under 3 MeV to test the drums of 320 ${\beta}\S$ and less; $(R)\ddot{E}$ both X-ray systems of 450 keV and/or 3 MeV might be needed considering the economical efficiency and the realization. The number of drums that can be tested with 450 keV and 3 MeV X-ray system was figured out as 42,327 and 18,105 drums (based on storage of 2006. 12), respectively. Four testing scenarios were derived considering equipment procurement method, outsourcing or not, etc. The economical and feasibility assessment for the scenarios was resulted in that an optimal scenario is dependent on the acceptance guide line, the waste generator's policy on the waste treatment and the delivery to a disposal facility, etc. For example, it might be desirable that a waste generator purchases two 450 keV mobile system to examine the drums containing low density waste, and that outsourcing examination for the high density drums, if all NDT items such as quantitative analysis for 'free standing water' and 'void fraction', and confirmation of contents in drum have to be characterized. However, one 450 keV mobile system seems to be required to test only the contents in 13,000 drums per year.

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Characterization of Burcucumber Biochar and its Potential as an Adsorbent for Veterinary Antibiotics in Water (가시박 유래 바이오차의 특성 및 항생물질 흡착제로서의 활용가능성 평가)

  • Lim, Jung Eun;Kim, Hae Won;Jeong, Se Hee;Lee, Sang Soo;Yang, Jae E;Kim, Kye Hoon;Ok, Yong Sik
    • Journal of Applied Biological Chemistry
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    • v.57 no.1
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    • pp.65-72
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    • 2014
  • Biochar (BC) from biomass pyrolysis is a carbonaceous material that has been used to remove various contaminants in the environment. The eliminatory action for burcucumber (Sicyos angulatus L.) as an invasive plant is being consistently carried out because of its harmfulness and ecosystem disturbance. In this study, burcucumber biomass was converted into BCs at different pyrolysis temperatures of 300 and $700^{\circ}C$ under a limited oxygen condition. Produced BCs were characterized and investigated to ensure its efficiency on antibiotics' removal in water. The adsorption experiment was performed using two different types of antibiotics, tetracycline (TC) and sulfamethazine (SMZ). For the BC pyrolyzed at a high temperature ($700^{\circ}C$), the values of pH, electrical conductivity, and the contents of ash and carbon increased whereas the yield, mobile matter, molar ratios of H/C and O/C, and functional groups decreased. Results showed that the efficiency of BCs on antibiotics' removal increased as pyrolysis temperature increased from 300 to $700^{\circ}C$ (38 to 99% for TC and 6 to 35% for SMZ). The reaction of ${\pi}-{\pi}$ EDA (electron-donor-acceptor) might be involved in antibiotics' adsorption to BCs. BC has potential to be a superior antibiotics' adsorbent with environmental benefit by recycling of waste/invasive biomass.

Mucin2 is Required for Probiotic Agents-Mediated Blocking Effects on Meningitic E. coli-Induced PathogenicitiesS

  • Yu, Jing-Yi;He, Xiao-Long;Puthiyakunnon, Santhosh;Peng, Liang;Li, Yan;Wu, Li-Sha;Peng, Wen-Ling;Zhang, Ya;Gao, Jie;Zhang, Yao-Yuan;Boddu, Swapna;Long, Min;Cao, Hong;Huang, Sheng-He
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1751-1760
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    • 2015
  • Mucin2 (MUC2), an important regulatory factor in the immune system, plays an important role in the host defense system against bacterial translocation. Probiotics known to regulate MUC2 gene expression have been widely studied, but the interactions among probiotic, pathogens, and mucin gene are still not fully understood. The aim of this study was to investigate the role of MUC2 in blocking effects of probiotics on meningitic E. coli-induced pathogenicities. In this study, live combined probiotic tablets containing living Bifidobacterium, Lactobacillus bulgaricus, and Streptococcus thermophilus were used. MUC2 expression was knocked down in Caco-2 cells by RNA interference. 5-Aza-2'-deoxycytidine (5-Aza-CdR), which enhances mucin-promoted probiotic effects through inducing production of Sadenosyl-L-methionine (SAMe), was used to up-regulate MUC2 expression in Caco-2 cells. The adhesion to and invasion of meningitic E. coli were detected by competition assays. Our studies showed that probiotic agents could block E. coli-caused intestinal colonization, bacteremia, and meningitis in a neonatal sepsis and meningitis rat model. MUC2 gene expression in the neonatal rats given probiotic agents was obviously higher than that of the infected and uninfected control groups without probiotic treatment. The prohibitive effects of probiotic agents on MUC2-knockdown Caco-2 cells infected with E44 were significantly reduced compared with nontransfected Caco-2 cells. Moreover, the results also showed that 5-Aza-CdR, a drug enhancing the production of SAMe that is a protective agent of probiotics, was able to significantly suppress adhesion and invasion of E44 to Caco-2 cells by upregulation of MUC2 expression. Taken together, our data suggest that probiotic agents can efficiently block meningitic E. coli-induced pathogenicities in a manner dependent on MUC2.

A Study of Hydrodynamic Dispersions in the Unsaturated and the Saturated Zone of a Multi-soil Layer Deposit Using a Continuous Injection Tracer Test (복합토양층의 불포화대와 포화대에서 연속주입 추적자시험을 이용한 수리분산특성 연구)

  • Chung, Sang-Yong;Kang, Dong-Hwan;Lee, Min-Hee;Son, Joo-Hyong
    • Journal of Soil and Groundwater Environment
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    • v.11 no.4
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    • pp.48-56
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    • 2006
  • Using a continuous injection tracer test at a multi-soil layer deposit, the difference of hydrodynamic dispersions in unsaturated and saturated zones were analyzed through breakthrough curves of Rhodamine WT, linear regression of concentration versus time, concentration variation rates versus time, and concentration ratio according to the distance from injection well. As a result of continuous injection tracer test, the difference of the maximum concentrations of Rhodamine WT in unsaturated and saturated zones were 13-15 times after 160 hours, and the increased rate of concentration versus time in unsaturated zone was about 10 times higher than in saturated zone. The fluctuation of Rhodamine WT breakthrough curve and concentration variation rate with time in saturated zone were larger than in unsaturated zone. Rhodamine WT concentration ratio with the distance from the injection well in saturation zone was linearly decreased faster than in unsaturated zone, and the elapsed time necessary for the concentration ratio less than 2 was longer in saturation zone. The differences resulted from the lower concentration and slower hydrodynamic dispersion of Rhodamine WT at the saturation zone of the multi-soil layer deposit, in which groundwater flow significantly flow and aquifer materials have high hydraulic heterogeneity. Effective porosity, longitudinal and transverse dispersivities were estimated $10.19{\sim}10.50%,\;0.80{\sim}1.98m$ and $0.02{\sim}0.04m$, respectively. The field longitudinal dispersivity is over 12 times larger than the laboratory longitudinal dispersivity by the scale-dependent effect.

Distribution and Ecology of Marsh Clam in Gyeongsangbuk-do II. Reproductive Cycle and Larval Development of the Corbicula japonica (경상북도 재첩자원 분포 및 생태 조사 II. 일본재첩 Corbicula japonica의 생식주기 및 유생발생)

  • 변경숙;정의영
    • The Korean Journal of Malacology
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    • v.17 no.1
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    • pp.45-55
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    • 2001
  • Gametogenes, reproductive cycle, first sexual maturity(biological minimum size), sex ratio and larval development of the marsh clam Corbicula japonica were investigated monthly by histological observations. Samples were collected in brackish water of Gokgang stream, Kyungsangbuk-Do, Korea, from August 1997 to July 1998. Sexuality of Corbicula japonica is dioecious and the species are an oviparous clam. The gonads are irregularly arranged from the sub-region of mid-intestinal gland in visceral cavity to reticular connective tissue of foot. The ovary is composed of a number of ovarian sac which are branched arborescent. Oogonia actively proliferate along the germinal epithelium of ovarian sac, in which young oocytes are growing. The testis is composed of a number of testicular tubules, and the epithelium of the tubule has function of germinal epithelium, along which spermatogonia actively proliferate. A great number of undifferentiated mesenchymal tissue and eosinophilic granular cells are abundantly distributed between developing oocytes and spermatocytes in the early developmental stages. With the further development of the ovary and testis these tissue and cells gradually disappear. Then the undifferentiated mesenchymal tissue and eosinophilic granular cells are considered to be related to the growing of the oocytes and spermatocytes. The spawning period is from July to September, and the main spawning occur between July and August when seawater temperatures reach above 22$^{\circ}C$. The reproductive cycle of this species can be divided into five successive stages; early active (February to April), late active (May to July), ripe (June to September), partially spawned (July to September), degenerative (September to October) and resting stage (October to February). Percentages of first sexual maturity of female and male clams ranging in length from 10 mm to 12 mm are over 50% and 100% for clams over 16.0 mm in shell length. Fertilized eggs or Corbicula japonica were 80-90 ${\mu}{\textrm}{m}$ in diameter. In the early embryonic development of C. japonica, the appearance of polar body, trochophore and D-shaped veliger were observed around 40 min., 27 hours and 4 days after spawning, respectively, at a water temperature of 26.5-28.$0^{\circ}C$. The size of larvae of early umbo stage was about 185-210 ${\mu}{\textrm}{m}$ in shell length, 160-180 ${\mu}{\textrm}{m}$ in shell height around 7 days after fertilization. The correlation of relative growth between the culture day (D) and shell length (SL) was expressed by the following simple formula from D-shaped veliger to metamorphosing stage; SL = 13.300D + 209.36($r^2$= 0.9078).

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Developing a Dental Unit Waterline Model Using General Laboratory Equipments (실험실 일반 장비를 이용한 치과용 유니트 수관 모델 개발)

  • Yoon, Hye Young;Lee, Si Young
    • Journal of dental hygiene science
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    • v.16 no.4
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    • pp.284-292
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    • 2016
  • Water supplied through dental unit waterlines (DUWLs) has been shown to contain high number of bacteria. To reduce the contamination of DUWLs, it is essential to develop effective disinfectants. It is, however, difficulty to obtain proper DUWL samples for studies. The purpose of this study was to establish a simple laboratory model for reproducing DUWL biofilms. The bacteria obtained from DUWLs were cultured in R2A liquid medium for 10 days, and then stored at $-70^{\circ}C$. This stock was inoculated into R2A liquid medium and incubated in batch mode. After 5 days of culturing, it was inoculated into the biofilm formation model developed in this study. Our biofilm formation model comprised of a beaker containing R2A liquid medium and five glass rods attached to DUWL polyurethane tubing. Biofilm was allowed to form on the stir plate and the medium was replaced every 2 days. After 4 days of biofilm formation in the laboratory model, biofilm thickness, morphological characteristics and distribution of the composing bacteria were examined by confocal laser microscopy and scanning electron microscopy. The mean of biofilm accumulation was $4.68{\times}10^4$ colony forming unit/$cm^2$ and its thickness was $10{\sim}14{\mu}m$. In our laboratory model, thick bacterial lumps were observed in some parts of the tubing. To test the suitability of this biofilm model system, the effectiveness of disinfectants such as sodium hypochlorite, hydrogen peroxide, and chlorhexidine, was examined by their application to the biofilm formed in our model. Lower concentrations of disinfectants were less effective in reducing the count of bacteria constituting the biofilm. These results showed that our DUWL biofilm laboratory model was appropriate for comparison of disinfectant effects. Our laboratory model is expected to be useful for various other purposes in further studies.

A study of the effects of PDGF-BB on the characteristics of bone stromal and periodontal ligament cells (혈소판유래성장인자-BB가 골간질세포와 치주인대세포의 성상에 미치는 영향)

  • Kwon, Young-Hyuk;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.949-965
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    • 1996
  • The main goal of periodontal therapy is to restore the lost periodontal tissue and establish the attachment appratus. Current acceptable therapeutic techniques are included : removal of diseased soft tissue, demineralization of exposed root surface, using the barrier membrane for preventing the downgrowth of gingival epithelial cell, insertion of graft materials as a scaffolding action, and biological mediators for promoting the cell activity. The latest concept one among them has been studied which based on the knowledge of cellular biology of destructed tissue. Platelet-derived growth factor(PDGF) is one of the polypeptide growth factor which have been reported as a biological mediator to regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the influences of the PDGF as biological mediator to periodontal ligament and bone marrow cell. Both right and left maxillary first molar were extracted from rat which had treated with 0.4% ${\beta}-Aminopropionitril$ for 5 days, and feeded until designed date to sacrifice under anesthesisa. Periodontal ligament were removed from the extracted socket of the rat, and cultured with Dulbecco's Modified Essential Medium(DMEM) contained with 10% Fetal Bovine Serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. Bone marrow cell were culture from bone marrow suspension with which washed out from femur with same medium. The study was performed to evaluate the effect of PDGF to periodontal ligament and bone cell, cell proliferation rate, total protein synthesis, and alkaline phosphatase activity of rat periodontal ligament(PDL) cell and bone stromal(RBS) cell in vitro. The effects of growth factors on both cells were measured at 3, 5th day after cell culture with (control group) or without growth factors(experimental group). The results were as follows: 1. The tendency of cell proliferation under the influence of PDGF showed more rapid proliferation pattern than control at 3 and 5 days after inoculation. 2. The activity of Alkaline phosphatase revealed 14, 80% increased respectively at 3, 5 days culture than control group. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(P<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P<0.05). 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group l(PDL 100%), 2(PDL 70%:GF 30%), and 3(PDL 50%:GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30%:GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05). From the above results, it is assumed that the PDGF on PDL cells and RMB cell culture. GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.

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Is Premedication Necessary for Outpatient Fiberoptic Bronchoscopy (외래환자의 기관지내시경검사시 전처치의 필요성)

  • Won, Jun-Hee;Park, Jae-Yong;Cha, Seung-Ick;Kang, Tae-Kyong;Park, Ki-Su;Kim, Yeon-Jee;Kim, Chang-Ho;Jung, Tee-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.2
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    • pp.251-259
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    • 1999
  • Background: Even though the necessity for premedication has been questioned, some combinations of antisialogogues, sedatives and analgesics are usually employed by most bronchoscopists. The goal of this study was to determine whether fiberoptic bronchoscopy(FOB) could be performed safely and effectively without premedication while using a standardized topical anesthetic. Method: Eighty outpatients were randomized in a double-blind manner into group I(n=20) with 1 ml normal saline, group II(n=20) with 0.5mg of atropine, group III(n=20) with 0.5mg of atropine plus 5mg of midazolam, and group IV(n=20) with 0.5mg of atropine plus 50mg of meperidine, given intramuscularly 30 minutes before FOB as premedication Topical anesthetics administered were same in each group. Each patient was given 5ml(200mg) of 4% nebulized lidocaine and additional intratracheobronchial 2% lidocaine. Oxygen saturation, pulse rate and electrocardiogram were monitored and recorded before, during, just after and 2 minutes after FOB. Immediately after FOB, the bronchoscopists answered four questions such as ease of procedure, extent of coughing, extent of secretion, and overall impression. Before leaving bronchoscopy suite, patients completed similar questions on discomfort during procedure, and willingness to return for a repeat procedure. Results: Age, gender, baseline pulmonary function, dose of 2% lidocaine used for topical anesthesia, and duration of FOB were not significantly different. There was no statistical difference among group I to IV with regard to extent of coughing answered by bronchoscopist. But extent of secretion was significantly different between group I without atropine and group II-IV with atropine. And there was also significant difference in ease of procedure and overall impression among groups. There was no statistical difference in patient's willingness and level of discomfort among the groups. Thirteen patients(16%) showed hypoxemia(arterial oxygen saturation: <90%), and 3 patients(4%) showed significant tachyarrhythmia(heart rate: > l60/min), but the rate of complication was not statistically different among the groups. Conclusion: These results suggest that the value of premedication is questionable for outpatient FOB, although it may be necessary in excessively anxious patient.

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Evaluation of the Clinical Usefulness of the Xeniss Rapid TB kit for the Diagnosis of Tuberculosis (결핵진단에서 Xeniss Rapid TB kit의 임상적 유용성)

  • Park, Seung-Kyu;Lee, Woo-Chul;Hwang, Soo-Hee;Kwon, Eun-Si;Lee, Hung-Soon;Lee, Duk-Hyoung
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.4
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    • pp.389-400
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    • 2002
  • Background : The rapid diagnostic tests for tuberculosis are needed to facilitate early treatment of tuberculosis and prevention of Mycobacterium tuberculosis transmission. The Xeniss Rapid TB kit is a rapid, card-based immunochromatographic test for the detection of antibodies directed against M. tuberculosis antigens including antigen 5(38-kDa antigen). The objective of this study was to evaluate the performance of the Xeniss Rapid TB kit for the diagnosis of active tuberculosis with serums from patients, asymptomatic healthy and close contact controls. Methods : 188 patients with active tuberculosis were tested; 177 with pulmonary tuberculosis(18 with combined pleurisy), and 11 with extrapulmonary tuberculosis. The control groups were composed of 82 close contacts and 57 healthy adults. Study subject were drawn from one national tuberculosis hospital for patients and close contacts, and another private hospital for healthy adults in Masan city, Korea. The Xeniss Rapid TB kit(Xeniss Life Science Co., Ltd., Seoul, Korea) was evaluated by using serum samples according to the instructions of the manufacturer by an investigator masked to the clinical and microbiological status of the study subjects. Results : The diagnostic sensitivity of the Xeniss Rapid TB kit was 73.9% in patients and specificities were 73.2% and 93.0% in close contact and healthy adults respectively. The positive predictive value in patients was 84.2% and the negative predictive value in controls was 85.8%. Conclusion : This study shows that the Xeniss Rapid TB test is a simple and fast method to diagnose active TB. The results of the sensitivity and specificites suggest that serodiagnosis using this point of care testing(POCT) device would be valuable and advantageous for screening tuberculosis in the clinical field.