DNA photography: An ultrasensitive DNA-detection method based on photographic techniques
DNA photography: An ultrasensitive DNA-detection method based on photographic techniques
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DOI：
10.1002/anie.200605023
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发表时间：
2007-01-01
影响因子：
16.6
通讯作者：
Carell, Thomas
中科院分区：
文献类型：
--
作者：
Hammond, David M.;Manetto, Antonio;Carell, Thomas
操纵和分析所有生物遗传信息的能力，以及小RNA链控制关键细胞功能的发现，促进了用于超灵敏检测DNA和RNA的新型复杂方法的发展在后基因组时代，人们相信这些技术将彻底改变基因编码疾病（如癌症）的诊断。事实上，更加个性化医疗的发展与可靠的超灵敏DNA检测方法的发展以及从生物样本中分离出感兴趣的基因（分析物）的新方法密切相关。本文所报道的方法为问题的第一部分提供了一个简单的解决方案。虽然聚合酶链反应（PCR）主要用于DNA检测，但高灵敏度检测DNA和RNA的新方法正在不断开发。例如，最近，Mirkin和他的同事、Willner和他的同事、Heeger和他的同事都报道了利用纳米粒子、适体或新型电化学装置来检测DNA，其灵敏度限制在微摩尔和飞摩尔范围内。[1-7]有报道称，在zeeptomolar溶液中（在30 μL中有% 10拷贝），甚至有敏感性所有这些方法都需要复杂的技术，这限制了它们的广泛使用。在此，我们报告了一种基于黑白摄影提供的扩增过程，在飞至原子（10À18 mol）范围内简单有效的DNA检测方法（图1）。在黑白摄影过程中，一些光子被含有agbr晶体的光敏层（胶片或相纸）捕获，诱导形成Agn核作为潜在图像中心。在随后的开发过程中，该Agn簇（对于n bbbb3）催化整个AgBr晶体还原为Ag0，使照相纸染黑迄今为止，基于银沉积的分析物的检测是用银和金团簇进行的，然后从溶液中沉积银，而不是通过照相纸溶液过程被限制在约105倍的放大倍数内。在此，我们报告了标准的和廉价的agbr晶体为基础的照相过程检测标记的生物分子的应用。该工艺原则上可提供与标准PCR反应相似的1011、[9]的扩增因子。
The ability to manipulate and analyze the genetic information of all organisms along with the discovery that small RNA strands control critical cellular functions has given rise to the development of new, sophisticated methods for the ultrasensitive detection of DNA and RNA.[1] In the postgenomic era, it is believed that such techniques will revolutionize the diagnosis of genetically encoded diseases such as cancer. In fact, the development of more-personalized medicine is critically linked to the development of reliable ultrasensitive DNA detection methods in combination with novel methods that allow isolation of the gene of interest (analyte) from biological samples. The method reported herein provides a simple solution to the first part of the problem. Although the polymerase chain reaction (PCR) is predominantly used for DNA detection, new methods for the highly sensitive detection of DNA and RNA are constantly being developed. Most recently, for example, Mirkin and co-workers, Willner and co-workers, and Heeger and co-workers have reported the utilization of nanoparticles, aptamers, or novel electrochemical setups to detect DNA with sensitivity limits in the range of pico-and femtomoles.[1–7] Even sensitivities in the zeptomolar regime (% 10 copies in 30 μL) have been reported.[8] All these methods require sophisticated technology, which limits their widespread use. We report herein a simple and efficient method for DNA detection in the femto-to attomole (10À18 mol) range based on the amplification process provided by black and white photography (Figure 1).In the black and white photographic process, a few photons captured by a light-sensitive AgBr-crystal-containing layer (film or photopaper) induce the formation of an Agn nucleus as a latent image center. In the subsequent development process, this Agn cluster (for n> 3) catalyzes the reduction of the entire AgBr crystal to Ag0, which stains the photopaper black.[9] To date, the detection of analytes based on silver deposition has been performed with Ag and Au clusters followed by Ag deposition from solution and not through photopaper.[10] The solution process is limited to amplification factors of about 105. Herein, we report the application of the standard and inexpensive AgBr-crystalbased photographic process for the detection of labeled biomolecules. This process can provide, in principle, amplification factors of 1011,[9] which are similar to standard PCR reactions.