Gen-Probe Incorporated, 10210 Genetic Center Drive, San Diego, California 92121, USA.
J Am Chem Soc. 2011 Sep 21;133(37):14637-48. doi: 10.1021/ja202221h. Epub 2011 Aug 26.
A novel method is described for simultaneous detection and quantification of attomoles or a few femtomoles of two (or potentially more) nucleic acid targets, without need for amplification. The technique depends on spectral-temporal resolution of chemiluminescence emitted from independent hybridization-induced chemiluminescent signal probes. The probes are internally quenched except in the presence of their specific targets, thereby allowing detection limits up to 10,000 times lower than with fluorescent probes. This is sufficient to obviate the need for amplification in many cases. The utility of the technique has been demonstrated by use of resolvable N-linked acridinium and 2,7-dimethoxyacridinium ester labeled probes in a homogeneous assay for sensitive and simultaneous independent quantification of pan-bacterial and pan-fungal target sequences in seawater.
描述了一种新的方法，用于同时检测和定量分析两个（或更多）核酸靶标，即使在没有扩增的情况下，也能达到飞摩尔或皮摩尔的检测水平。该技术依赖于化学发光的光谱-时间分辨率，这种化学发光是由独立的杂交诱导化学发光信号探针发出的。这些探针在没有特定目标存在的情况下会被内部猝灭，从而使检测限比荧光探针低 10000 倍。在许多情况下，这足以避免对扩增的需求。该技术的实用性已通过在海水样本中使用可分辨的 N 连接吖啶盐和 2,7-二甲氧基吖啶酯标记探针的均相测定中得到了证明，可用于敏感且同时独立地定量分析泛细菌和泛真菌目标序列。
