Thin film biosensor for rapid visual detection of nucleic acid targets.
Thin film biosensor for rapid visual detection of nucleic acid targets.
复制标题
用于快速视觉检测核酸靶标的薄膜生物传感器。
作者：
R. Ostroff;Deborah Hopkins;A. Haeberli;Wahab Baouchi;B. Polisky
背景
我们已经开发了一种基于硅的生物传感器，其响应于核酸靶标而产生视觉信号。
方法
在该系统中，捕获寡核苷酸探针固定在生物传感器的表面上。捕获探针与互补靶和生物素化的检测寡核苷酸的相互作用允许在生物传感器上开始形成有机薄膜。薄膜形成是通过与抗生物素抗体结合的过氧化物酶的酶活性完成的。过氧化物酶催化不溶性产物在硅表面上的沉积，产生均匀的薄膜。表面上增加的厚度通过改变来自表面的反射光的干涉图案来改变生物传感器的感知颜色，导致颜色从金色变为紫色。
结果
生物传感器结果可以通过直接目视检查来评估或通过椭圆光度法来量化。在25分钟内获得结果，检测限为5 pmol/L（150阿莫尔/样品）。通过对单核苷酸错配的区分证明了生物传感器的选择性。多目标阵列也与薄膜生物传感器进行了分析，该系统能够检测来自人血清和尿液的目标。
结论
生物传感器表面生产成本低廉，并且测定形式简单快速。薄膜生物传感器适用于各种核酸检测应用，包括传染病组、抗生素抗性组或特定遗传标记的等位基因鉴别的快速诊断测试。
BACKGROUND
We have developed a silicon-based biosensor that generates a visual signal in response to nucleic acid targets.
METHODS
In this system, capture oligonucleotide probes are immobilized on the surface of the biosensor. Interaction of the capture probes with a complementary target and a biotinylated detector oligonucleotide allows initiation of formation of an organic thin film on the biosensor. Thin film formation is completed by enzymatic activity of peroxidase conjugated to an anti-biotin antibody. Peroxidase catalyzes deposition of an insoluble product onto the silicon surface, generating a uniform thin film. The increased thickness on the surface alters the perceived color of the biosensor through changes in the interference patterns of reflected light from the surface, causing a color change from gold to purple.
RESULTS
The biosensor results may be evaluated by direct visual inspection or quantified by ellipsometry. Results are obtained in 25 min with a detection limit of 5 pmol/L (150 amol/sample). Selectivity of the biosensor is demonstrated by discrimination of single nucleotide mismatches. Multitarget arrays are also analyzed with the thin film biosensor, and the system is capable of detecting targets from human serum and urine.
CONCLUSIONS
The biosensor surface is inexpensive to produce, and the assay format is simple and rapid. The thin film biosensor is adaptable to a wide variety of nucleic acid detection applications, including rapid diagnostic testing for infectious disease panels, antibiotic resistance panels, or allelic discrimination of specific genetic markers.
影响因子：
3.3
作者：
MANSFIELD, ES;WORLEY, JM;FORTINA, P
通讯作者：
FORTINA, P
影响因子：
56.9
作者：
Lin, VSY;Motesharei, K;Ghadiri, MR
通讯作者：
Ghadiri, MR
影响因子：
56.9
作者：
Elghanian, R;Storhoff, JJ;Mirkin, CA
通讯作者：
Mirkin, CA