A dual-signal amplification rapid detection platform integrating ERA and lateral flow immunoassay.

In the field of point-of-care testing (POCT), the rapid and sensitive detection of Mycoplasma pneumoniae is essential for the early diagnosis and prognosis. However, current testing methods are primarily qualitative and require sophisticated instrumentation. Herein, we present a POCT platform that integrates enzymatic recombinase amplification (ERA) and microsphere lateral flow strips (LFS) with high-fluorescence intensity, employing a dual-signal amplification approach to achieve rapid and sensitive detection of Mycoplasma pneumoniae, which is called the Smartphone-based ERA-LFS Integrated Device (SELID). Concurrently, smartphone readout software has been developed to realize image enhancement, as well as the recognition of control and test lines via designed algorithms for quantitative analysis and subsequent data processing. The platform can complete the assay within 25 min, achieving high specificity and sensitivity, with the ability to detect as low as 10 copies/μL. In addition, we added Mycoplasma pneumoniae strains to saliva to simulate clinical samples to verify the detection ability of SELID in real samples. Experimental findings demonstrated strong concordance between SELID and the conventional PCR assay, exhibiting good feasibility and stability. These results indicate that SELID has significant potential as a simple, rapid, and sensitive diagnostic platform for the immediate detection of Mycoplasma pneumoniae and other pathogens.