Integrated detection method for separating, enriching and detecting urine exosome as well as detection chip
An integrated detection and exosome technology, applied in the field of biotechnology detection, can solve the problems of inability to meet the detection requirements and low exosome content, and achieve the effect of easy acquisition and great application prospects
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Embodiment 1
[0053] Example 1: Separation of exosomes in urine of bladder cancer patients and detection by chip ELISA method, refer to the attached Figure 1-8 .
[0054] see Figure 1-7 , an integrated detection method for urine exosome isolation, enrichment and ELISA detection, comprising the following steps:
[0055] 1) On the premise of the patient’s knowledge and the consent of the ethics committee, 16 urine samples were collected from bladder cancer patients (clinical biopsy confirmed bladder cancer), 8 healthy donors, each 100ml, and centrifuged at 2,000g at room temperature Cells and large debris were removed, and the supernatant was obtained.
[0056] 2) Ultracentrifuge the supernatant obtained in 1) at 100,000 g for 60 minutes at 4°C, discard the supernatant to collect the precipitate, and resuspend the exosomes with 1 mL of PBS buffer with pH 7.4 (such as figure 1 ).
[0057] 3) Inject the supernatant obtained in 1) into the double-membrane filter chip at a flow rate of 40 μ...
Embodiment 2
[0066] Embodiment 2, integrated detection chip, with reference to attached Figure 1-3 .
[0067] The microfluidic chip of the present invention comprises a base plate 1, a top plate 2, and a detection plate located between the base plate and the top plate, the detection plate is provided with a liquid detection cavity, and the liquid detection cavity can remove exosomes in urine It is collected in it and enriched by multiple injection samples to meet the detection requirements.
[0068] The detection board includes an upper detection board 3 and a lower detection board 4. The top board 2 is provided with a liquid injection hole 5 and a liquid discharge hole 12 formed by cutting, and the liquid injection hole 5 is connected with a liquid injection pipe 6. The liquid discharge hole 12 is connected with a liquid discharge pipe 13, and the upper layer detection plate 3 and the lower layer detection plate 4 are respectively provided with a sample injection hole 7 and a waste liqu...
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