Integrated paper-based detection chip with nucleic acid extraction and amplification for automatic and sensitive pathogen detection

Abstract

Molecular diagnostics with high sensitivity and specificity are essential for the early detection of infectious diseases caused by pathogens. However, it has not reached its full potential in point-of-care diagnosis because of the technically demanding and labor-intensive methods, which limit its accessibility in low-resource areas. Herein, we propose an integrated genetic analysis platform that fully automates sample pretreatment, nucleic acid amplification and endpoint detection. Magnetic silica beads were employed to isolate DNA from lysate to simplify the device design and operation. A helical continuous-flow polymerase chain reaction (PCR) reactor was divided into three sections with different temperatures to perform the denaturing, annealing and extension phases of the PCR. Additionally, the final detection was performed using a nucleic acid paper-based detection chip, and the results were automatically analyzed by software. Using this device, as few as 104 CFU/mL of Listeria monocytogenes can be detected. This device prevents cross-contamination, greatly reduces the number of operation steps, and improves detection efficiency. Further, it has the potential to be utilized for rapid decision making related to pathogen containment and to eliminate the requirements for end-users with professional skills and specialized laboratory facilities.