Nucleic acid sample preparation from whole blood in a paper microfluidic device using isotachophoresis

Promise and perils of paper-based point-of-care nucleic acid detection for endemic and pandemic pathogens

“…Gan et al 63 demonstrated extraction of DNA from whole blood, dried blood spots, and saliva among other samples by alternating, high and low pH of rinsing buffers followed by water elution. Sullivan et al 64 used fusion 5 in a handheld membrane-based device for blood fractionation. The authors used isotachophoresis (ITP), an electrokinetic extraction technique, coupling with membrane filtrations to remove leukocytes and erythrocytes from whole blood and concentrate eluted nucleic acids.…”

Progression of LAMP as a Result of the COVID-19 Pandemic: Is PCR Finally Rivaled?

“…A significant obstacle to genetic identification at the point-of-care sites is initial DNA extraction, as the process is lengthy and requires skilled personnel to conduct lysis of the cell membrane, isolation and purification of the DNA, and various washing steps [ 42 , 43 ]. Some available techniques to speed up and simplify conventional RNA-purification and DNA-extraction methods include: microfluidic chips with cartridges dedicated to purification steps [ 44 , 45 , 46 , 47 ], bead- or membrane-based purification [ 48 , 49 , 50 ], and magnetic particles [ 51 , 52 , 53 ], which can bypass any centrifugation through components that directly bind to DNA/RNA for isolation [ 42 ]. These can speed up the process and lessen the amount of trained personnel needed.…”

Quantitative isothermal amplification on paper membranes using amplification nucleation site analysis

“…We have developed novel chemistry and paper-based cell filtration and nucleic acid extraction techniques that are compatible with the point-of-care and avoid the use of chaotropic salts. 63,64 Viral detection in nasal and saliva samples, such as SARS-CoV-2 testing, can be accomplished at the POC with minimal sample preparation by direct addition of the sample into viral transport media with a lysis surfactant and reducing agent (such as DTT). 51,65,66 Integration of sample preparation with the presented quantitative nucleic acid amplification is the focus of our future work.…”