Simple, sensitive and accurate multiplex detection of clinically important melanoma DNA mutations in circulating tumour DNA With SERS nanotags

Wee, Eugene J. H., Wang, Yuling, Simon, Chang-Hao Tsao and Trau, Matt (2016) Simple, sensitive and accurate multiplex detection of clinically important melanoma DNA mutations in circulating tumour DNA With SERS nanotags. Theranostics, 6 10: 1506-1513. doi:10.7150/thno.15871


Author Wee, Eugene J. H.
Wang, Yuling
Simon, Chang-Hao Tsao
Trau, Matt
Title Simple, sensitive and accurate multiplex detection of clinically important melanoma DNA mutations in circulating tumour DNA With SERS nanotags
Journal name Theranostics   Check publisher's open access policy
ISSN 1838-7640
Publication date 2016-06-17
Year available 2016
Sub-type Article (original research)
DOI 10.7150/thno.15871
Open Access Status DOI
Volume 6
Issue 10
Start page 1506
End page 1513
Total pages 8
Place of publication Ivyspring International Publisher
Publisher Sydney, NSW Australia
Language eng
Abstract Sensitive and accurate identification of specific DNA mutations can influence clinical decisions. However accurate diagnosis from limiting samples such as circulating tumour DNA (ctDNA) is challenging. Current approaches based on fluorescence such as quantitative PCR (qPCR) and more recently, droplet digital PCR (ddPCR) have limitations in multiplex detection, sensitivity and the need for expensive specialized equipment. Herein we describe an assay capitalizing on the multiplexing and sensitivity benefits of surface-enhanced Raman spectroscopy (SERS) with the simplicity of standard PCR to address the limitations of current approaches. This proof-of-concept method could reproducibly detect as few as 0.1% (10 copies, CV < 9%) of target sequences thus demonstrating the high sensitivity of the method. The method was then applied to specifically detect three important melanoma mutations in multiplex. Finally, the PCR/SERS assay was used to genotype cell lines and ctDNA from serum samples where results subsequently validated with ddPCR. With ddPCR-like sensitivity and accuracy yet at the convenience of standard PCR, we believe this multiplex PCR/SERS method could find wide applications in both diagnostics and research.
Keyword SERS
Multiplex PCR
Melanoma
BRAF
cKIT
NRAS
ctDNA
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: HERDC Pre-Audit
Australian Institute for Bioengineering and Nanotechnology Publications
 
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Citation counts: TR Web of Science Citation Count  Cited 5 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 8 times in Scopus Article | Citations
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Created: Tue, 05 Jul 2016, 01:14:32 EST by Eugene Wee on behalf of Aust Institute for Bioengineering & Nanotechnology