RNA quantification using gold nanoprobes - application to cancer diagnostics.
RNA quantification using gold nanoprobes - application to cancer diagnostics.
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使用金纳米探针进行 RNA 定量——在癌症诊断中的应用。
DOI：
10.1186/1477-3155-8-5
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发表时间：
2010-02-24
影响因子：
10.2
通讯作者：
Baptista PV
中科院分区：
文献类型：
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作者：
Conde J;de la Fuente JM;Baptista PV
应用于癌症的分子纳米诊断可以提供对癌症相关分子改变的快速和灵敏的检测，这将使得即使当这些改变仅发生在一小部分细胞中时也能够早期检测。使用巯基修饰的寡核苷酸（Au-纳米探针）衍生的金纳米颗粒用于检测特定的核酸靶标作为更传统的方法的替代方法已经获得了发展势头。在这里，我们提出了一种基于Au纳米颗粒的方法，用于BCR-ABL融合转录物（mRNA）的分子识别和定量，这是慢性粒细胞白血病（CML）的原因，据我们所知，这是第一次直接在癌细胞中定量特定的mRNA。这种廉价且非常容易执行的基于Au纳米探针的方法允许定量未扩增的总人类RNA和癌基因转录物的特异性检测。通过Au纳米探针确定的灵敏度允许从10 ng/μl总RNA的差异基因表达，并且在总RNA提取后需要不到30分钟完成，从而最大限度地减少RNA降解。此外，在晚期，恶性突变的积累可能导致对化疗的耐药性，从而导致不良结局。这种方法，允许嵌合BCR-ABL mRNA的快速和直接的检测和定量，可以加快诊断，并在适当的情况下，修改治疗。该检测可能构成CML早期诊断的有前途的工具，并且可以容易地扩展到被证明参与癌症发展的进一步靶基因。
Molecular nanodiagnostics applied to cancer may provide rapid and sensitive detection of cancer related molecular alterations, which would enable early detection even when those alterations occur only in a small percentage of cells. The use of gold nanoparticles derivatized with thiol modified oligonucleotides (Au-nanoprobes) for the detection of specific nucleic acid targets has been gaining momentum as an alternative to more traditional methodologies. Here, we present an Au-nanoparticles based approach for the molecular recognition and quantification of the BCR-ABL fusion transcript (mRNA), which is responsible for chronic myeloid leukemia (CML), and to the best of our knowledge it is the first time quantification of a specific mRNA directly in cancer cells is reported. This inexpensive and very easy to perform Au-nanoprobe based method allows quantification of unamplified total human RNA and specific detection of the oncogene transcript. The sensitivity settled by the Au-nanoprobes allows differential gene expression from 10 ng/μl of total RNA and takes less than 30 min to complete after total RNA extraction, minimizing RNA degradation. Also, at later stages, accumulation of malignant mutations may lead to resistance to chemotherapy and consequently poor outcome. Such a method, allowing for fast and direct detection and quantification of the chimeric BCR-ABL mRNA, could speed up diagnostics and, if appropriate, revision of therapy. This assay may constitute a promising tool in early diagnosis of CML and could easily be extended to further target genes with proven involvement in cancer development.