时间隔离可以提高能量转移测定中的灵敏度,这些测试采用了terbium 酸盐/暗灭器寡核酸探针
Mary Katherine Johansson1, Ronald M Cook, Jide Xu
1Biosearch Technologies, 81 Digital Drive, Novato, California 94949, USA. marykat@biosearchtech.com
Journal of the American Chemical Society
|December 17, 2004
概括
带有暗灭剂的兰他尼德复合物增强了寡核酸探针的灵敏度. 时间隔离检测在探针切割后显著放大了发光信号,改善了测试性能.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 分析化学 分析化学
背景情况:
- 兰化物提供长时间的发光衰变,用于时隔检测,区分信号和背景噪声.
- 复合物与四硫胺 (TIAM) 合物提供稳定,发光的生物标签.
- 多芳香-阿佐暗灭器 (黑洞灭器,BHQ) 有效地灭了兰坦化物发光.
研究的目的:
- 开发和描述使用 terbium/TIAM 复合物和 BHQ 火器的双标记的寡核酸探针.
- 为了研究 terbium-BHQ 距离和发光灭效率之间的关系.
- 为了证明这些探测器的增强灵敏度,在能量转移试验中使用时隔检测.
主要方法:
- 合成了5'TIAM (Tb)/3'BHQ双标记的寡核酸探针.
- 在完整和裂开的探针中测量发光强度和寿命.
- 稳定状态与时隔检测方法对信号放大进行比较.
主要成果:
- 发光灭高度依赖于特复合体和BHQ之间的距离.
- 探测器裂变 (杂交或核酶消化) 在空间上将Tb和BHQ分开,增加发光.
- 与稳定状态检测 (增加3倍) 相比,时隔检测提供了显著的信号增强 (例如,40-mer探测器的50倍增强).
结论:
- /暗灭器寡核酸探针通过发光能量转移进行敏感检测.
- 时隔检测是一种强大的策略,可以放大信号并提高这些测试中的灵敏度.
- 这种方法为各种生物和化学检测应用提供了可靠的方法.
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