▎ 摘　　要

NOVELTY - Chip comprises functional substrate, flow passage layer and valve control layer, which are orderly arranged from bottom to top. The functional substrate is divided into graphene oxide (GO) nucleic acid capture area and polylysine (PLL) detection area. A multiple detecting units are arranged on flow channel layer. Each detecting unit includes micro-cavity one, sample inlet one, and sample outlet one that penetrate flow channel layer up and down. A sample inlet flow passage and sample outlet flow passage opened on back of channel layer. The one end of sample inlet flow passage is connected to sample inlet one, and other end is connected to microcavity one. The one end of sample outlet flow passage is connected to sample inlet flow passage, and other end is connected to sample outlet one. The sample flow channel is located above GO nucleic acid capture area on functionalized substrate. USE - The chip is useful in kit for detecting novel coronavirus nucleic acid (all clamed). ADVANTAGE - The chip: is economical; and has simple device. The kit: has high detection sensitivity, less sample demand, simple detection process and short time; and does not require extraction and amplification of nucleic acid. DETAILED DESCRIPTION - Chip comprises functional substrate, flow passage layer and valve control layer, which are orderly arranged from bottom to top. The functional substrate is divided into graphene oxide (GO) nucleic acid capture area and polylysine (PLL) detection area. A multiple detecting units are arranged on flow channel layer. Each detecting unit includes micro-cavity one, sample inlet one, and sample outlet one that penetrate flow channel layer up and down. A sample inlet flow passage and sample outlet flow passage opened on back of channel layer. The one end of sample inlet flow passage is connected to sample inlet one, and other end is connected to microcavity one. The one end of sample outlet flow passage is connected to sample inlet flow passage, and other end is connected to sample outlet one. The sample flow channel is located above GO nucleic acid capture area on functionalized substrate. The microcavity is located in PLL detection area on functionalized substrate. The capture probe E of E segment of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus is immobilized on functionalized substrate in region of injection channel. A valve control layer is provided with multiple micro-cavity two, sample inlet two and sample outlet two which penetrate valve control layer up and down. The position of second micro-cavity corresponds to position of first micro-cavity. The second sample inlet corresponds to first sample inlet. The second sample outlet corresponds to first sample outlet. The valve control layer is further provided with valve inlet one and valve inlet two that penetrate valve control layer up and down. The back of valve control layer is provided with control flow channel one connected with valve inlet one and control flow channel two connected with valve inlet two. A micro-cavity control valve is arranged above inlet position of each micro-cavity one on control flow channel one. A micro-channel control valve is arranged on control flow channel two above position of each sample outlet flow channel. INDEPENDENT CLAIMS are included for: (1) preparing the chip, comprising (i) preparing functionalized substrate, preparing of flow channel layer and preparing valve control layer, (ii) placing the prepared valve control layer on the upper part of the flow channel layer, butting together under the microscope, baking at 70degreesC for a period of time, cooling to room temperature, and placing on a functionalized substrate to prepare a chip, and (iii) immobilizing of the capture probe E of the E segment of the SARS-CoV-2 virus; (2) a kit for detecting novel coronavirus nucleic acid, comprising the chip, a blocking solution, a cleaning solution, a lysis reagent, a negative control substance and a positive control substance, the chip is used to detect SARS-CoV-2 virus nucleic acid, the blocking solution is used to occupy the site of the un-immobilized capture probe E during the detection process, the cleaning solution cleans the functionalized substrate on which the DNA-RNA double strands have been fixed at the end of the detection, the lysis reagent is used to process the SARS-CoV-2 virus to obtain RNA, using the negative control substance for qualitative comparison with the result of the sample to be tested to confirm whether there is SARS-CoV-2 virus in the sample to be tested, and using positive control substance to quantitatively analyze the RNA content of the SARS-CoV-2 virus in the sample to be tested; and (3) use of the kit in detecting novel coronavirus nucleic acid, comprising (1) drawing of the standard curve by injecting four different concentrations of positive controls into the four microcavities of the chip, incubating for a period of time, incubating the whole chip in 3% BSA, peeling off the flow channel layer and valve control layer, soaking the functionalized substrate for 10 minutes, using the cleaning solution to clean the functionalized substrate, finally drying the functionalized substrate for fluorescence detection, analyzing the fluorescence value, and performing linear fitting with the concentration to make a standard curve, (2) processing virus samples to obtain RNA by adding the lysis reagent to the virus sample, placing the whole container containing the above mixed solution in the ice-water mixture, and ultrasonically processing ice-water mixture by a cell disruptor to obtain a sample solution, (3) qualitatively detecting of RNA in virus samples by the sample solution and injecting negative control substance into the blank micro-chamber one of the chip, incubating for a period of time, immersing the whole chip in 3% BSA, removing flow channel layer and valve control layer, soaking the functionalized substrate for 10 minutes, using the cleaning solution to clean the functionalized substrate, finally drying the functionalized substrate as for fluorescence detection, comparing the fluorescence value of the sample with the fluorescence value of the negative control, and determining whether the sample solution contains novel coronavirus, and (4) quantitatively detecting RNA in virus samples by injecting the sample solution into the blank micro-cavity one of the chip, incubating for a period of time, immersing the whole chip in 3% BSA, peeling off the flow channel layer and valve control layer, soaking the functionalized substrate for 10 minutes, using the cleaning solution to clean the functionalized substrate, finally drying the functionalized substrate for fluorescence detection, and obtaining the RNA content in the sample according to the standard curve.