Fully integrated nucleic acid pretreatment, amplification, and detection on a paper chip for identifying EGFR mutations in lung cancer cells

Paper-based diagnostics have been successfully explored for nucleic acid detection. However, multi-step integration (such as nucleic acid extraction, washing, amplification and detection) into a single paper chip remains a huge challenge. Here, a simple, inexpensive, and fully integrated paper chip was demonstrated for detecting gene mutations, which could potentially play a guiding role in individualized treatments to improve the survival rate of lung cancer patients. To simplify the pipetting and manipulation of samples, we incorporated DNA extraction, purification, amplification and detection into a single chip. Further integration of lateral flow test allowed the detection of gene mutation by naked eye. The gene mutation in the epidermal growth factor receptor (EGFR(L858R)) in lung adenocarcinoma cells was successfully detected with high specificity and sensitivity within 90 min by the proposed fully integrated chip. This low cost, disposable and user-friendly chip provided a new and rapid molecular diagnostic platform, especially suitable for molecular diagnostics in resource-limited areas.