Fig. 6: On-site detection of MP using POP-CRISPR.

a Workflow of POP-CRISPR for on-site testing for MP using mini-device. Green arrows and red numbers (①–③) indicate the three main steps and their sequential order in the detection workflow. First, nasopharyngeal swab clinical samples are collected and mixed with Chelex-100-containing lysis buffer, followed by rapid nucleic acid release on the mini-device at 80 °C for 2 min. The obtained nucleic acids are then added to the POP-CRISPR reaction system, which is performed on the mini-device using a poikilothermal one-pot protocol (37 °C for 10 min followed by 45 °C). The detection results can be interpreted via wireless connection to a smartphone application, serving as a reference for subsequent diagnosis and prognosis. Selected icons in this figure were created by soco-st and are available on SVGRepo.com (https://www.svgrepo.com/svg/492696/online-medical-treatment-female; https://www.svgrepo.com/svg/493281/fever-female). These icons are licensed under the Creative Commons Attribution 4.0 International License (CC BY 4.0) (https://creativecommons.org/licenses/by/4.0/), in accordance with the SVGRepo licensing terms (https://www.svgrepo.com/page/licensing/#CC%20Attribution). b Scheme for standardized nucleic acid extraction and rapid lysis for nucleic acid release. c Evaluation the extraction efficiency of two nucleic acid extraction methods using POP-CRISPR. 27 clinical nasopharyngeal swab samples suspected of MP infection were tested. Values are shown as endpoint fluorescence at 60 min. The threshold value (red dashed line) was set based on the mean fluorescence of no target control plus three times the standard deviation. d Real-time fluorescence profiles obtained from POP-CRISPR experiments performed on mini-device. The experimental procedure was as follows: 10 min at 37 °C followed by 45 °C, figure shows the real-time fluorescence signal of 45 °C. F.I. represents fluorescence intensity. a.u. represents arbitrary units. Source data are provided as a Source Data file.