MilliporeSigma
  • Highly sensitive DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization.

Highly sensitive DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization.

Biosensors & bioelectronics (2014-12-17)
Xu Yu, Zhi-Ling Zhang, Si-Yang Zheng
ABSTRACT

A novel highly sensitive colorimetric assay for DNA detection using cascade amplification strategy based on hybridization chain reaction and enzyme-induced metallization was established. The DNA modified superparamagnetic beads were demonstrated to capture and enrich the target DNA in the hybridization buffer or human plasma. The hybridization chain reaction and enzyme-induced silver metallization on the gold nanoparticles were used as cascade signal amplification for the detection of target DNA. The metalization of silver on the gold nanoparticles induced a significant color change from red to yellow until black depending on the concentration of the target DNA, which could be recognized by naked eyes. This method showed a good specificity for the target DNA detection, with the capabilty to discriminate single-base-pair mismatched DNA mutation (single nucleotide polymorphism). Meanwhile, this approach exhibited an excellent anti-interference capability with the convenience of the magentic seperation and washing, which enabled its usage in complex biological systems such as human blood plasma. As an added benefit, the utilization of hybridization chain reaction and enzyme-induced metallization improved detection sensitivity down to 10pM, which is about 100-fold lower than that of traditional unamplified homogeneous assays.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Trizma® hydrochloride, anhydrous, free-flowing, Redi-Dri, ≥99.0%
Supelco
Glutathione, Pharmaceutical Secondary Standard; Certified Reference Material
Glutathione, European Pharmacopoeia (EP) Reference Standard
SAFC
MES hydrate
Trolamine impurity B, European Pharmacopoeia (EP) Reference Standard
Supelco
Diethanolamine, analytical standard
Sigma-Aldrich
Trizma® hydrochloride, BioUltra, for molecular biology, ≥99.0% (AT)
Sigma-Aldrich
MES monohydrate, BioXtra, ≥99.0% (T)
Sigma-Aldrich
MES monohydrate, BioUltra, for molecular biology, ≥99.5% (T)
Sigma-Aldrich
Diethanolamine, reagent grade, ≥98.0%
Sigma-Aldrich
MES hydrate, ≥99.5% (titration), pH 2.5-4.0 (0.5 M in H2O), BioXtra
Sigma-Aldrich
Diethanolamine, BioUltra, ≥99.5% (GC)
Sigma-Aldrich
MES hydrate, ≥99.5% (titration)
Sigma-Aldrich
MES hydrate, BioUltra, ≥99.5% (T)
Sigma-Aldrich
Trizma® hydrochloride, reagent grade, ≥99.0% (titration), crystalline
Sigma-Aldrich
Trizma® hydrochloride, pH 3.5-5.0 (0.5 M in H2O), BioXtra, ≥99.0% (titration)
Sigma-Aldrich
Trizma® hydrochloride, BioPerformance Certified, suitable for cell culture, ≥99.0% (titration)
Sigma-Aldrich
MES hydrate, BioPerformance Certified, suitable for cell culture, ≥99.5%
Sigma-Aldrich
Sodium borohydride solution, ~12 wt. % in 14 M NaOH
Sigma-Aldrich
Diethanolamine, ACS reagent, ≥98.5%
Sigma-Aldrich
Sodium borohydride solution, 2.0 M in triethylene glycol dimethyl ether
Sigma-Aldrich
Diethanolamine, puriss. p.a., ACS reagent, ≥99.0% (GC)
SAFC
MES monohydrate
SAFC
TRIS hydrochloride
Sigma-Aldrich
VenPure® SF, powder
Sigma-Aldrich
1,2-Dichloroethane, ACS reagent, ≥99.0%
Sigma-Aldrich
L-Glutathione reduced, suitable for cell culture, BioReagent, ≥98.0%, powder
Supelco
1,2-Dichloroethane, analytical standard
Sigma-Aldrich
Sodium borohydride, granular, 99.99% trace metals basis
Sigma-Aldrich
MES solution, BioUltra, for molecular biology, 0.5 M in H2O