A single-molecule Raman detection test kit and its usage method are disclosed. The test kit includes a test strip slot on one side and a sample injection mechanism on the other side. The test strip slot is inclined, and a test strip with a pointed end is placed inside the slot. The tip of the test strip is positioned at the lowest point of the slot, and the
tail end is positioned at the highest point. The sample injection mechanism includes a channel, the bottom of which is located above the highest point of the slot, and a spiral sampler is placed inside the channel. The usage method involves rinsing the spiral sampler with deionized water, allowing it to dry, and then dripping the
analyte solution into the spiral sampler. Driven by the components of gravity and capillary force, the liquid is transported along the inclined test strip slot to the tip of the test strip obtained by a
polymer-assisted printing method for adsorption. The surface and gaps of Au@Ag NPs clusters are filled at the tip of the test strip, completing the enhanced enrichment of the
analyte molecules. After the test kit is ventilated and dried, SERS measurement is performed and quantitative analysis is conducted. This invention utilizes the Au@Ag NPs at the tip of the test strip obtained by
polymer-assisted printing. The combination of NPs clusters and the enhanced tip enrichment method used in the test strip kit enables high-sensitivity applications, achieving 10 in practical sample applications. ‑ 9 The
detection limit of M enables detection at the single-molecule level.