He Jing, Zhu Jianjun, Gong Changguo, Qi Jiming, Xiao Han, Jiang Bin, Zhao Yulan
School of Life Science, East China Normal University, Shanghai, PR China.
Ocean Research Center of Zhoushan, Zhejiang University, Zhejiang Province, PR China.
PLoS One. 2015 Dec 28;10(12):e0145160. doi: 10.1371/journal.pone.0145160. eCollection 2015.
The diagnostic and prognostic value of microRNAs (miRNAs) in a variety of diseases is promising. The novel silicon nanowire (SiNW) biosensors have advantages in molecular detection because of their high sensitivity and fast response. In this study, poly-crystalline silicon nanowire field-effect transistor (poly-SiNW FET) device was developed to achieve specific and ultrasensitive detection of miRNAs without labeling and amplification.
The poly-SiNW FET was fabricated by a top-down Complementary Metal Oxide Semiconductor (CMOS) wafer fabrication based technique. Single strand DNA (ssDNA) probe was bind to the surface of the poly-SiNW device which was silanated and aldehyde-modified. By comparing the difference of resistance value before and after ssDNA and miRNA hybridization, poly-SiNW device can be used to detect standard and real miRNA samples.
Poly-SiNW device with different structures (different line width and different pitch) was applied to detect standard Let-7b sample with a detection limitation of 1 fM. One-base mismatched sequence could be distinguished meanwhile. Furthermore, these poly-SiNW arrays can detect snRNA U6 in total RNA samples extracted from HepG2 cells with a detection limitation of 0.2 μg/mL. In general, structures with pitch showed better results than those without pitch in detection of both Let-7b and snRNA U6. Moreover, structures with smaller pitch showed better detection efficacy.
Our findings suggest that poly-SiNW arrays could detect standard and real miRNA sample without labeling or amplification. Poly-SiNW biosensor device is promising for miRNA detection.
微小RNA(miRNA)在多种疾病中的诊断和预后价值前景广阔。新型硅纳米线(SiNW)生物传感器因其高灵敏度和快速响应在分子检测方面具有优势。在本研究中,开发了多晶硅纳米线场效应晶体管(poly-SiNW FET)器件,以实现对miRNA的特异性和超灵敏检测,无需标记和扩增。
采用基于自上而下的互补金属氧化物半导体(CMOS)晶圆制造技术制造poly-SiNW FET。将单链DNA(ssDNA)探针结合到经过硅烷化和醛修饰的poly-SiNW器件表面。通过比较ssDNA与miRNA杂交前后电阻值的差异,poly-SiNW器件可用于检测标准和实际miRNA样本。
应用具有不同结构(不同线宽和不同间距)的poly-SiNW器件检测标准Let-7b样本,检测限为1 fM。同时可以区分单碱基错配序列。此外,这些poly-SiNW阵列能够检测从HepG2细胞中提取的总RNA样本中的小核RNA U6,检测限为0.2 μg/mL。总体而言,在检测Let-7b和小核RNA U6时,有间距结构的检测结果优于无间距结构。而且,间距较小的结构检测效果更好。
我们的研究结果表明,poly-SiNW阵列无需标记或扩增即可检测标准和实际miRNA样本。Poly-SiNW生物传感器器件在miRNA检测方面具有广阔前景。