DNA Detection Using a Triple Readout Optical/AFM/MALDI Planar Microwell Plastic Chip

Abstract

A ready-to-spot disposable DNA chip for specific and sensitive detection of DNA was developed. Plastic copolymeric substrate chemistry was optimized to selectively couple the target DNA with the active chip surface. At the same time, the developed substrate limits the unspecific adsorption of probe DNA molecules or additional polar contaminants in the test samples to the chip surface. The combination of glycidyl and n-butyl methacrylates was found to best fit the requirements of the assay. The fabricated DNA microarrays have mechanical properties similar to those of the glass or silicon substrates and, at the same time, provide chemically reactive surfaces that do not require lengthy chemical modification. An additional advantage of the plastic microchip is its compatibility with different analytical readout techniques, such as mass spectrometry (MALDI-TOF/MS), optical detection (fluorescence and enzyme-induced metal deposition), and imaging techniques (atomic force microscopy). These multiple readout techniques have given us the ability to compare the sensitivity, selectivity, and robustness of current state-of-the-art bioanalytical methods on the same platform exemplified by successful DNA-based detection of human cytomegalovirus. The obtained sensitivity for enzymatically enhanced silver deposition (10(-15) M) surpasses that of conventional fluorescence readouts. In addition, the assay's dynamic range (10(-6)-10(-15) M), reproducibility, and reliability of the DNA probe detection speaks for the silver deposition method. At compromised sensitivity (10(-9) M), the length of the DNA probes could be checked and, alternatively, DNA single point polymorphisms could be analyzed.

A microfluidic platform for chip-based DNA detection using SERS and silver colloids

“…Appropriate materials for the chip platform are glass, polymers or hydrogels. 26,27 Here, we use glass chips for the immobilisation of the capture DNA. For the DNA analysis, different capture DNA strands were immobilised on the chip substrate.…”

Microbial forensics: fiber optic microarray subtyping of Bacillus anthracis

“…Microarrays are most definitely being employed for new and interesting problems and techniques, including AFM, Raman, and electrochemical analyses. [17][18][19][20][21][22][23] The applications to which these combination techniques are being used are well outside the initial scope or originally intended functions associated with basic method. More succinctly, microarrays are broadening other techniques into previously unlikely applications.…”