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    • 5. 发明授权
    • IS6110 based molecular detection of mycobacterium tuberculosis
    • IS6110基因分子检测结核分枝杆菌
    • US5731150A
    • 1998-03-24
    • US551645
    • 1995-11-01
    • Gurpreet S. SandhuBruce C. KlineLeslie StockmanGlenn D. RobertsMarcia E. Lewis
    • Gurpreet S. SandhuBruce C. KlineLeslie StockmanGlenn D. RobertsMarcia E. Lewis
    • C12N15/09C12Q1/04C12Q1/68C12R1/32C07H21/00C07H21/04C12P19/34
    • C12Q1/689
    • The detection of the IS6110 insertion element in a clinical specimen is a rapid way of diagnosing infection by Mycobacterium tuberculosis. A reliable diagnostic test for tuberculosis based on the IS6110 DNA is described in this disclosure. A "Universal" specimen preparation protocol that eliminates live organisms and purifies nucleic acids from all types of clinical specimens is described. Two nucleic acid primers designed to amplify IS6110 DNA with high specificity in a polymerase chain reaction are also described. The amplified IS6110 DNA is identified by a restriction endonuclease and electrophoresis based assay. The identification process also renders the DNA unamplifiable in a subsequent PCR, thereby reducing the possibility of contaminating other specimens. Time, labor and cost is minimized, while user safety and test reliability are maximized. The complete DNA extraction, amplification and analysis is accomplished with ease within an 8 hour period, with a sensitivity of 92% and a specificity approaching 100%. Testing of serially obtained samples from the same patient increases the overall rate of detection to 100%.
    • 临床标本中IS6110插入元件的检测是诊断结核分枝杆菌感染的一种快速方法。 本公开描述了基于IS6110 DNA的结核病的可靠诊断测试。 描述了消除活生物体并从所有类型的临床标本中纯化核酸的“通用”标本制备方案。 还描述了设计用于在聚合酶链反应中以高特异性扩增IS6110DNA的两个核酸引物。 通过限制性内切酶和基于电泳的测定法鉴定扩增的IS6110 DNA。 识别过程也使DNA在随后的PCR中不可扩增,从而降低污染其他样品的可能性。 时间,人力和成本最小化,用户安全和测试可靠性最大化。 完整的DNA提取,扩增和分析在8小时内容易地完成,灵敏度为92%,特异性接近100%。 来自同一患者的连续获得的样品的测试将总体检测率提高到100%。