Direct and rapid detection of RNAs on a novel RNA microchip.
Direct and rapid detection of RNAs on a novel RNA microchip.
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DOI：
10.1002/cbic.201000170
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发表时间：
2010-07-05
期刊：
影响因子：
3.2
通讯作者：
Huang, Zhen
中科院分区：
文献类型：
--
作者：
Spencer, Sarah M.;Lin, Lina;Chiang, Cheng-Feng;Peng, Zhengchun;Hesketh, Peter;Salon, Jozef;Huang, Zhen
Direct, rapid, and accurate detection of pathogen RNAs is essential for controlling the spread of infectious diseases, such as H1N1 influenza or other infectious agents that can cause epidemics.[1-3] Nucleic acid-based detection systems, such as real-time PCR and DNA microchips (or microarrays), offer high detection specificity and sensitivity and are able to detect single-nucleotide differences in pathogens, genes, and diseases.[4-14] Though these technologies are powerful, RNAs from viable pathogens are not directly detected, and must be indirectly analyzed by cDNA amplification and detection. It is also difficult to use these technologies to detect the partially degraded pathogenic RNAs that can provide information on pathogen viability. Since it is difficult to amplify and label RNAs, it is extremely challenging to directly detect RNAs, let alone fragments. In addition, a few of the current methods for RNA direct detection, such as the Northern blot, RNase protection assay, microRNA profiling, and direct RNA sequencing,[15] are labor intensive, costly or instrument-intensive. They are not well suited to rapid and high-throughput RNA detection, especially not for point-of-care diagnosis and other applications.The direct and rapid detection of RNA remains an unsolved challenge. To face this problem, our research group has developed a methodology for accurate RNA detection by taking advantage of nuclease and polymerase activities. Herein we report for the first time the proof of principle of a novel RNA microchip strategy for the direct and rapid detection of RNAs (Scheme 1). Our simple strategy is fast (ca. 20 min detection time) and of high specificity (single-nucleotide discrimination) and high sensitivity (the low-femtomole level). Furthermore, we have successfully demonstrated the selective detection of multipathogen RNAs in a synthetic mixture and an individual mRNA in total RNA from a biological sample.
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