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    • 4. 发明公开
    • 실시간 역전사효소 중합반응을 이용한 결핵의 진단을 위한 정보제공방법 및 이를 위한 결핵 진단용 키트
    • 提供使用定量实时PCR诊断结核病的信息的方法及其包含的诊断包
    • KR1020120103387A
    • 2012-09-19
    • KR1020110033865
    • 2011-04-12
    • 연세대학교 원주산학협력단
    • 이혜영김성현조상래김태우
    • C12Q1/68C12N15/11
    • C12Q1/689C12Q2600/158G01N33/50G01N33/96
    • PURPOSE: A kit for diagnosing tuberculosis by real-time RT-PCR is provided to enable test using a small amount of blood. CONSTITUTION: A method for providing information for diagnosing tuberculosis comprises: a step of collecting blood from a patient who is suspicious for tuberculosis and treating tuberculosis bacillus-specific antigen to the blood; a step of isolating full length RNA from the blood; a step of synthesizing cDNA from the full length RNA; a step of performing real-time RT-PCR; and a step of measuring IFN-gamma mRNA expression level. The tuberculosis bacillus-specific antigen is ESAT-6, CFP-10 or TB7.7. A primer for amplifying IFN-gamma contains a primer pair of sequence numbers 1 and 2. [Reference numerals] (AA) Step of collecting blood from a patient who is suspected to have tuberculosis and treating with tuberculosis bacillus-specific antigen; (BB) Step of isolating full-length RNA from blood treated with tuberculosis bacillus-specific antigen; (CC) Step of synthesizing cDNA from the isolated full length RNA; (DD) Step of real time RT-PCR of synthesized cDNA using first primer pair which is able to amplify IFN-γ, first probe which binds to amplified site, second primer pair which is able to amplify control group gene to compare with IFN-γ expression level, and second probe which binds to amplified control group gene; (EE) Step of measuring mRNA expression level of the amplified IFN-γ and mRNA expression level of control group
    • 目的:提供用于通过实时RT-PCR诊断结核病的试剂盒,以便使用少量血液进行检测。 构成:提供用于诊断结核病的信息的方法包括:从可疑结核病患者中收集血液并将血液结核杆菌特异性抗原治疗的步骤; 从血液中分离全长RNA的步骤; 从全长RNA合成cDNA的步骤; 进行实时RT-PCR的步骤; 以及测量IFN-γmRNA表达水平的步骤。 结核杆菌特异性抗原是ESAT-6,CFP-10或TB7.7。 用于扩增IFN-γ的引物含有序列号1和2的引物对。(附图标记)(AA)从怀疑患有结核病并用结核杆菌特异性抗原治疗的患者收集血液的步骤; (BB)从结核杆菌特异性抗原处理的血液中分离全长RNA的步骤; (CC)从分离的全长RNA合成cDNA的步骤; (DD)使用能够扩增IFN-γ的第一引物对的合成cDNA的实时RT-PCR步骤,与扩增位点结合的第一个探针,能够扩增对照组基因以与IFN- γ表达水平,与扩增对照组基因结合的第二个探针; (EE)测量扩增的IFN-γ的mRNA表达水平和对照组的mRNA表达水平的步骤