Paper-based detection method for exosome
A detection method and technology of exosomes, which are used in the determination/inspection of microorganisms, biochemical equipment and methods, etc.
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Embodiment 1
[0063] Example 1 Establishment of a paper-based detection method for exosomes
[0064] 1. Experimental materials
[0065] 20×SSC, Tris-HCl, sodium phosphate dodecahydrate, bovine serum albumin (BSA), Tween-20, sucrose and TMB chromogenic solution, etc.
[0066] The sequence design is as follows (SEQ ID NO:1-4):
[0067]
[0068] 2. LFB detection and verification of single-stranded DNA targets
[0069] Dilute the single-stranded DNA target 2-fold concentration gradient, add 50 µL system buffer and 1 µL single-stranded DNA target solution to the sample pool, and drop-coat 1 µL PtAu-NiO on the superposition of the test paper sample pad and the binding pad 2 After the nanozyme signal probe, insert the test paper sample pad into the sample pool, wait for 3 minutes, and observe the result with the naked eye. When both the C line and the T line are black, it is determined to be positive. The color of the T line becomes lighter as the concentration of the ssDNA target decreases,...
Embodiment 2
[0078] Example 2 Practical investigation of detection method
[0079] According to the detection method constructed in Example 1, exosomes extracted from equal volumes of HepG2 and L02 cell supernatants were detected. LFB can effectively distinguish tumor exosomes from non-tumor exosomes ( Image 6 ).
Embodiment 3
[0080] Embodiment 3 Amplification effect verification of detection method
[0081] After conversion and amplification of exosomes, the gray pixel area of each T line in the LFB detection results was substituted into the gray pixel area of the T line obtained by LFB detection after 2-fold gradient dilution of the single-stranded DNA target to perform standard curve fitting. Formula: y =14.829e 0.0005x In , the concentration of the single-stranded DNA target after exosome conversion and amplification was obtained. The exosome concentration was converted to the concentration of the substance and compared with the single-stranded DNA target. The molecular magnification of exosomes by this method is 1000 times (Table 1).
[0082] Table 1 Molecular magnification of exosomes after target conversion and amplification
[0083]
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