An internal amplification control for quantitative nucleic acid analysis using nanoparticle-based dipstick biosensors
An internal amplification control for quantitative nucleic acid analysis using nanoparticle-based dipstick biosensors
复制标题
使用基于纳米粒子的试纸生物传感器进行定量核酸分析的内部放大控制
DOI：
10.1016/j.bios.2012.10.078
复制
发表时间：
2013-04-15
影响因子：
12.6
通讯作者：
Zhou, Guohua
中科院分区：
文献类型：
--
作者：
Huang, Huan;Jin, Li;Zhou, Guohua
病毒核酸的定量分析对于基于血液中病毒RNA或DNA的拷贝数来监测医学治疗的功效是必不可少的。为了定量检测血液中的病毒核酸，本文提出了一种与基于纳米颗粒的DNA生物传感器耦合的内部扩增对照（IAC）。设计具有特定序列的IAC，并在核酸提取之前掺入血清中。IAC和靶标的序列仅在一个PCR引发位点的碱基顺序上不同;因此，IAC和靶标的Tm相同，从而在PCR期间产生相同的扩增效率。为了视觉检测扩增子，采用基于链霉亲和素官能化纳米颗粒的试纸生物传感器。通过比较生物传感器上测试区与IAC区的颜色密度，可以半定量地分析血清中靶标的含量。这种方法已经实现了在大约100个拷贝的病原体载量下检测HBV DNA。该方法的可行性证明了成功的半定量病原体载量在30例HBV感染患者的临床样本。这些数据表明，IAC和基于纳米颗粒的试纸型生物传感器的引入可能是护理点测试（POCT）中的有力工具。(C)2012爱思唯尔有限公司版权所有。
Quantitative analysis of virus nucleic acids is essential for monitoring the efficacy of medical treatment based on the copy numbers of virus's RNA or DNA in blood. To quantitatively detect virus nucleic acids in blood, here an internal amplification control (IAC) coupled with a nanoparticle-based DNA biosensor was proposed. The IACs with a specific sequence were designed and spiked into serum before nucleic acids extraction. Sequences of the IACs and the targets only differ in the base order of one PCR priming site; thus, the IACs and the targets are identical in Tm, giving the same amplification efficiency during PCR. To visually detect amplicons, a dipstick biosensor based on streptavidin-functionalized nanopartides is employed. By comparing color densities of a test zone with an IAC zone on the biosensor, the content of the target in serum can be semi-quantitatively analyzed. This approach has achieved the detection of HBV DNA at approximately 100 copies of the pathogen load. The feasibility of this method is demonstrated by successful semi-quantification of pathogen load in 30 clinical samples from HBV-infected patients. These data indicate that the introduction of an IAC and nanoparticle-based dipstick-type biosensor could be a powerful tool in point of care testing (POCT). (C) 2012 Elsevier B.V. All rights reserved.