结核分支杆菌分子信标探针荧光芯片固定方式的研究

Fix of the fluorescence array of molecular beacon probe of tuberculosis bacillus

目的研究结核分支杆菌分子信标探针荧光芯片的固定方式。方法通过对传统生物素(biotin)-亲和素(avidin)连接方式和自行设计的单侧延长臂的分子信标连接方式的对比研究,对TB分子信标与芯片固定方式进行探索。结果生物素-亲和素(biotin-avidin)能很好地结合在芯片上但这种修饰有相当技术难度;且生物素-亲和素连接无特异性。本设计单侧延长臂分子信标采用淬灭分子中间标记,茎结构单侧(淬灭分子一侧)延长臂,能很好的结合在芯片上,且修饰技术难度低、特异性高。结论单侧延长臂分子信标探针设计的发明及运用,初步解决了分子信标技术在芯片技术领域运用的固定难题。巧妙地把分子信标高特异性、高灵敏性等优势自然运用到芯片技术上。

Objective To research on the fix of the fluorescence array of bacillus tuberculosis molecular beacon probe. Methods Traditional biotin-avidin connection and unilateral extended arm connection were compared for fixing TB molecular beacon probe to the chip. Hybridization efficiency of molecular beacon probe with unilateral extended arm on the chip was optimized. Results Unilateral extended arm molecular beacon with quenching molecular middle label and stem structure unilateral extended arm （one side was quenched） could be easily fixed onto the chip and specifically modified. Conclusions The design and apply of unilateral extended arm molecular beacon probe initially solved the difficulty in molecular beacon technology applied on chip technology.