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Chemical Engineering Journal

Chemical Engineering Journal

IF: 13.3
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Bisulfite-free detection of DNA methylation in early-stage lung cancer with a multiplex SERS immunoassay

Published:10 December 2024 DOI: 10.1016/j.cej.2024.158491
Dechan Lu, Yanping Chen, Longfeng Ke, Zufang Huang, Yudong Lu, Jing Wang

Abstract

DNA methylation is a promising biomarker for early cancer detection; however, traditional detection methods require bisulfite treatment and nucleic acid amplification, which are costly and impractical for point-of-care use. We propose a facile method that leverages immunoassays and surface-enhanced Raman scattering (SERS) readouts for the multiplex, sensitive, and portable detection of lung cancer-related DNA methylation biomarkers. This approach employs 5-methylcytosine antibody-modified magnetic probes to enrich methylated DNA sequences and uses core–shell nanoparticles with Probe DNA as SERS nanotags for multiplex detection, eliminating the need for bisulfite treatment and nucleic acid amplification. The multiplex SERS immunoassay detects synthetic methylated SHOX2 and RASSF1A with detection limits of 0.52 pM and 0.66 pM, respectively. It shows a strong correlation with quantitative PCR results in analyzing methylated SHOX2 and RASSF1A from cell lines and formalin-fixed paraffin-embedded tissue samples (n?=?35). Additionally, the random forest analysis based on methylated SHOX2 and RASSF1A expression distinguishes lung cancer from benign lung diseases with a clinical sensitivity of 96?%, specificity of 90?%, and an AUC value of 0.972. This method offers a significant advancement for DNA methylation research and clinical applications.

Substances (11)

Materials
Procduct Name CAS Molecular Formula Supplier Price
Ascorbic Acid 50-81-7 C6H8O6 1626 suppliers $6.00-$2170.00
Ethylene glycol 107-21-1 C2H6O2 1400 suppliers $10.00-$908.54
Sodium chloride 7647-14-5 ClNa 1214 suppliers $5.00-$6678.62
Sodium hydroxide 1310-73-2 NaOH 1211 suppliers $14.00-$16472.00
L-Glutamine 56-85-9 C5H10N2O3 1140 suppliers $5.00-$10696.00
Monoethanolamine 141-43-5 C2H7NO 943 suppliers $9.00-$4290.00
Polyvinylpyrrolidone 9003-39-8 CH4 809 suppliers $15.00-$9201.92
Ethanol 64-17-5 C2H6O 806 suppliers $10.00-$17310.00
Proteinase K 39450-01-6 C29H27N2O12P 408 suppliers $45.48-$9570.00
POLYETHYLENEIMINE 9002-98-6 C2H5N 363 suppliers $29.20-$2864.00