Highly sensitive and homogeneous detection of unamplified RNA based on the light scattering properties of gold nanoparticle probes
Highly sensitive and homogeneous detection of unamplified RNA based on the light scattering properties of gold nanoparticle probes
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DOI：
10.1016/j.biosx.2022.100249
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发表时间：
2022-12
影响因子：
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通讯作者：
H. Satoh;M. Nakajima;Reiko Hirano;Yuki Nakaya
中科院分区：
文献类型：
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作者：
H. Satoh;M. Nakajima;Reiko Hirano;Yuki Nakaya
RNA是几种人类疾病的重要诊断生物标志物。因此，用于检测病原体来源的RNA（例如，RNA病毒）在水中是转录组学分析所必需的。在这里，我们提出了一个高度敏感的检测方法，以检测未扩增的RNA的基础上的光散射特性的荧光肽功能化的金纳米粒子探针（金纳米探针）。在该测定中，Au-纳米探针识别靶RNA并与靶RNA杂交。随后，当样品溶液被放置在石英波导载玻片上并被倏逝波激发时，它们散射光。该检测试剂盒可在RNA提取后10 min内对细菌16 S rRNA和mRNA进行高灵敏度定量（约102拷贝/μL），无需逆转录和cDNA扩增。此外，该测定比先前报道的用于直接RNA定量的测定灵敏三个数量级。
RNA is a crucial diagnostic biomarker for several human diseases. Hence, a simple, rapid, sensitive, and cost-effective assay for detecting pathogen-derived RNAs (e.g., RNA viruses) in water is necessary for transcriptomic analysis. Here, we present a highly sensitive assay to detect unamplified RNA based on the light scattering properties of oligonucleotide-functionalized gold nanoparticle probes (Au-nanoprobes). In this assay, Au-nanoprobes recognize and hybridize to the target RNA. Subsequently, they scatter light when the sample solution is placed onto a quartz waveguide slide and is excited with an evanescent wave. This assay enables highly sensitive quantification (approximately 102copies/μL) of bacterial 16S rRNA and mRNA without reverse transcription and cDNA amplification within 10 min of RNA extraction. Moreover, this assay was three orders of magnitude sensitive than previously reported assays for direct RNA quantification.