MP77-05 A SIMPLE BUT HIGH EFFECTIVE SPECIMEN PREPARATION FOR DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS AND ITS CLINICAL APPLICATION IN QUANTITATIVE-PCR DETERMINATION OF URINARY TUBERCULOSIS INFECTION

You have accessJournal of UrologyInfections/Inflammation/Cystic Disease of the Genitourinary Tract: Kidney & Bladder II (MP77)1 Apr 2020MP77-05 A SIMPLE BUT HIGH EFFECTIVE SPECIMEN PREPARATION FOR DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS AND ITS CLINICAL APPLICATION IN QUANTITATIVE-PCR DETERMINATION OF URINARY TUBERCULOSIS INFECTION Zhuo Hui, Cao Min*, He Ben, Yuan Renbin, and Guo Yuanbiao Zhuo HuiZhuo Hui More articles by this author , Cao Min*Cao Min* More articles by this author , He BenHe Ben More articles by this author , Yuan RenbinYuan Renbin More articles by this author , and Guo YuanbiaoGuo Yuanbiao More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000963.05AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: In the recent few years, there is an ongoing outbreak of urinary tuberculosis worldwide. Molecular diagnosis based on genomic amplification is a rapid and reliable method for early detection of tuberculosis infection. However, the quality and quantity of nucleic acid of Mycobacterium tuberculosis (MTB) extracted from urine is still a bottleneck for molecular analysis because of the difficulty associated with breaking cell walls of MTB. METHODS: In this study, new specimens handling methods including Dounce homogenizer with a loose pestle and a tight pestle, compared with a conventional procedure consisted of sodium hydroxide, were applied for DNA isolation from urine with or without MTB, and real time quantitative PCR (RT-qPCR) were evaluated DNA recovery, sensitivity and specifisicty for early and accurate diagnosis of tuberculosis infection. RESULTS: 45 patients infected with urinary tuberculosis and 56 normal controls were enrolled. With specimens handling with both the tight pestle and loose pestle of Dounce homogenizer, MTB DNA was recovered about 300 -104 times than traditional specimen pretreated method and 83.67%-100% tuberculosis infections could be better detected. The new DNA extraction methods were also performed excellent in urine with detection rate in the range of 85.71%-90.47%. The relative lower specificity (80.55%-92.31%) was also well explained by follow-up. RT-qPCR handling with D-Tight data expanding with another 209 suspicious TB individuals (164 diagnosed with TB infection and 45 diagnosed with non-TB infection) showed a high sensitivity (97.47%, taken clinical diagnosis as golden standard) with no significant diverse between TB treated group and TB non-treated group (96.30% VS 98.08%). CONCLUSIONS: The results demonstrates that Dounce homogenizer could offer high quality and quantity MTB DNA to RT-qPCR determination of urinary MTB infection. Source of Funding: This work was supported by National Natural Science Foundation of China (Jin: Grant No. 81370799 and Cao: Grant No. 31500077); Science and Technology Support Program of Sichuan Province (Grant No. 2015FZ0077 and 14ZC0926); Narural Science Foundation of Zhejiang Province (Jin: Grant No. LGF18H050001 and Chen: LY16H050002). © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e1164-e1165 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Zhuo Hui More articles by this author Cao Min* More articles by this author He Ben More articles by this author Yuan Renbin More articles by this author Guo Yuanbiao More articles by this author Expand All Advertisement PDF downloadLoading ...