Analytical method for identifying prostate cancer based on urine exosomes
A prostate cancer and analysis method technology, which is applied in the analysis field of prostate cancer identification based on urine exosomes, can solve the problems of low accuracy rate, high re-examination rate, incompatibility of PCR instruments, etc., and achieve the effect of high accuracy rate
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Embodiment 1
[0033] An analysis method for identifying prostate cancer based on urine exosomes, comprising the steps of:
[0034] S1. Collect 10 mL of urine from the patient at the initial stage, centrifuge at 4°C and 2500 rpm for 5 minutes, discard the supernatant, add 1.0 mL of PBS solution to the bottom sediment, and repeatedly blow and mix to obtain a mixed exosome solution;
[0035] S2. Add 1.3mL Trizol solution to the exosome solution in S1, let stand for 3min, add 0.2mL chloroform, mix upside down, centrifuge at 2°C and 12000rpm for 13min, take the upper clear solution, and pass the upper clear solution through several centrifuges, Obtain sample RNA solution;
[0036] S3. Take 5 μL each of the positive quality control product, the negative quality control product, and the sample RNA solution in S2, respectively, and mix them with RT-PCR amplification reagents (the specific RT-PCR amplification reagents include PSA, PCA3, ERG, and SPDEF). Primers and probes) were amplified to obtain...
Embodiment 2
[0057] An analysis method for identifying prostate cancer based on urine exosomes, comprising the steps of:
[0058] S1. Collect 12 mL of urine from the patient at the initial stage, centrifuge at 2°C and 2200 rpm for 7 minutes, discard the supernatant, add 1.2 mL of LPBS solution to the bottom sediment, and repeatedly blow and mix to obtain a mixed exosome solution;
[0059] S2. Add 1.0mL Trizol solution to the exosome solution in S1, let it stand for 4min, add 0.4mL chloroform, mix it upside down, centrifuge at 4°C and 11000rpm for 12min, take the upper clear solution, and pass the upper clear solution through several centrifuges, Obtain sample RNA solution;
[0060] S3. Take 6 μL each of the positive quality control product, the negative quality control product, and the sample RNA solution in S2, and mix them with RT-PCR amplification reagents (the specific RT-PCR amplification reagents include PSA, PCA3, ERG, and SPDEF). Primers and probes) were amplified to obtain positi...
Embodiment 3
[0081] An analysis method for identifying prostate cancer based on urine exosomes, comprising the steps of:
[0082] S1. Collect 14mL of urine from the patient at the initial stage, centrifuge at 4°C and 2000rpm for 10min, discard the supernatant, add 1.4mL of PBS solution to the bottom sediment, and repeatedly blow and mix to obtain a mixed exosome solution;
[0083] S2. Add 1.5mL Trizol solution to the exosome solution in S1, let it stand for 5min, add 0.6mL chloroform, mix it upside down, centrifuge at 4°C and 11000rpm for 15min, take the upper clear solution, and pass the upper clear solution through multiple centrifuges, Obtain sample RNA solution;
[0084] S3. Take 7 μL each of the positive quality control product, the negative quality control product, and the sample RNA solution in S2, respectively, and mix them with RT-PCR amplification reagents (the specific RT-PCR amplification reagents include PSA, PCA3, ERG, and SPDEF). Primers and probes) were amplified to obtain...
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