Journal
ANALYTICAL CHEMISTRY
Volume 94, Issue 37, Pages 12860-12865Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.2c02894
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Funding
- Guangdong Basic and Applied Basic Research Foundation [2022A1515010618]
- Guangzhou Science and Technology Project [202201010600, 202201020170]
- National Natural Science Foundation of China [21904026, 21974031]
- Innovation Training Program for College Students of Guangzhou University [202211078113, S202111078031]
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Polymers have great potential as carriers for signal tags in amplified detection. This study presents a polysaccharide-amplified method for electrochemical detection of a specific DNA target. The method involves attaching carboxyl group containing polysaccharides to a peptide nucleic acid capture probe, followed by the decoration of polysaccharide chains with electroactive ferrocene signal tags. The method shows high sensitivity, selectivity, and cost-effectiveness.
In view of their high efficiency and cost-effectiveness, polymers are of great promise as carriers for signal tags in amplified detection. Herein, we present a polysaccharide-amplified method for the electrochemical detection of a BRCA1 breast cancer gene-derived DNA target at the femtomolar levels. Briefly, peptide nucleic acid (PNA) with a complementary sequence was tethered as the capture probe for the DNA target, to which carboxyl group containing polysaccharides were then attached via facile phosphateZr(IV)-carboxylate crosslinking, followed by the decoration of polysaccharide chains with electroactive ferrocene (Fc) signal tags via affinity coupling between a cis-diol site and phenylboronic acid (PBA) group. As the polysaccharide chain contains hundreds of cisdiol sites, boronate affinity can enable the site-specific decoration of each polysaccharide chain with hundreds of Fc signal tags, efficiently transducing each target capture event into the decoration of many Fc signal tags. As polysaccharides are cheap, renewable, ubiquitous, and biodegradable natural biopolymers, the use of polysaccharides for signal amplification offers the benefits of high efficiency, cost-effectiveness, excellent biocompatibility, and environmental friendliness. The linear range of the polysaccharide amplified method for DNA detection was demonstrated to be from 10 fM to 10 nM (R-2 = 0.996), with the detection limit as low as 2.9 fM. The results show that this method can also discriminate single base mismatch with satisfactory selectivity and can be applied to DNA detection in serum samples. In view of these merits, the polysaccharide-amplified PNA-based electrochemical method holds great promise in DNA detection with satisfactory sensitivity and selectivity.
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