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      저자 : ( Chul-ho Oack ) (검색결과 1 건)
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      • Thresholds value of transrenal DNA for the diagnosis of pulmonary tuberculosis and treatment monitoring

        ( Taeyoon Kim ),( Hyun Kyung Kwak ),( Chul-ho Oack ),( Jehun Kim ),( Taeyun Kim ),( Sungweon Ryoo ) 대한결핵 및 호흡기학회 2019 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.127 No.-

        There have been attempts to use biological samples other than sputum for the diagnosis of TB. Urine is promising clinical specimen because of its availability, ease of access, processing and storage, and the low infection risk to healthcare workers during sample collection. In this study, we evaluated the thresholds of trans-renal (tr) DNA detection in patients with MDR, XDR pulmonary tuberculosis compared with artificially synthesized Mycobacterium tuberculosis (M.TB)-specific DNA. During the study cell free DNA urine preserve was added quickly in order to minimize degradation of soluble DNA by nucleases and adapted highly efficient isolation method for circulating cell free DNA isolation. We designed artificial double strand of M.TB bacilli and used it as control DNA. The artificially synthesized control DNA, trDNA extracted from M.TB H37Rv strain and patients’ urine were serially diluted and then the copy number were calculated using droplet digital PCR system. The quantitative value of M.TB-specific trDNA extracted from 10 MDR, XDR TB patients were confirmed by gene amplification technology. The detection efficiencies of our trDNA isolation outcomes were compared and quantify the total fraction, an average yield of >10-4 pg/㎕ was detected. Urine samples are increasingly used for TB diagnosis. However, extraction and concentration of trDNA is necessary for many molecular detection strategies. Since urine samples typically have large volumes with dilute M.TB specific trDNA concentrations making them prone to false negatives. In this study, we established a method of extracting trDNA from urine and a control DNA standard that can quantify very small amounts of trDNA. The detection of M.TB trDNA from urine specimen is a promising method not only for the diagnosis TB but also to assess treatment responses.

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