Somodevilla-Torres M, Timms P, Harris R, Morris C P, Van Daal A
Cooperative Research Centre for Diagnostic Technologies, Queensland University of Technology, Brisbane, Qld 4001, Australia.
Mol Diagn. 2001 Jun;6(2):131-6. doi: 10.1054/modi.2001.24436.
We report the development of an enzyme-linked immunosorbent assay-like single-tube assay for the detection of infectious agents in a microtiter tray format.
The method, sequential nucleic acid amplification and capture (SNAAC), combines amplification with hybridization of the product to a surface/matrix-bound oligonucleotide probe. After amplification of the target sequence using species-specific primers, one of which contains a detection tag such as fluorescein or biotin, a denaturation and hybridization cycle is performed. This allows capture by an oligonucleotide that is covalently bound to the surface of a microtiter tray well or other support. After washing to remove unincorporated solution-phase oligonucleotide bearing the detection tag, the level of captured product is determined through a colorimetric reaction using an automated plate reader. We show the value and utility of the SNAAC detection method using cloned sequences of the important human respiratory pathogen Chlamydia pneumoniae.
SNAAC is a simple, rapid, and inexpensive method for the detection of low levels of infectious agents that is readily adaptable to current clinical laboratory equipment, thus avoiding the need to develop or purchase new instrumentation.
我们报告了一种类似酶联免疫吸附测定的单管检测方法的开发，该方法以微量滴定板形式检测感染因子。
该方法即序列核酸扩增与捕获（SNAAC），将扩增与产物与表面/基质结合的寡核苷酸探针杂交相结合。使用物种特异性引物扩增靶序列后，其中一个引物含有诸如荧光素或生物素等检测标签，进行变性和杂交循环。这允许通过与微量滴定板孔表面或其他支持物共价结合的寡核苷酸进行捕获。洗涤以去除未结合的带有检测标签的溶液相寡核苷酸后，使用自动酶标仪通过比色反应确定捕获产物的水平。我们使用重要的人类呼吸道病原体肺炎衣原体的克隆序列展示了SNAAC检测方法的价值和实用性。
SNAAC是一种简单、快速且廉价的检测低水平感染因子的方法，很容易适应当前临床实验室设备，从而无需开发或购买新仪器。
