Kit for detecting cancer prognosis
A detection kit and detection reagent technology, applied in the field of medicine and health, can solve the problems of limited kidney cancer and the need to enhance the prediction effect of the model, and achieve the effect of high predictive value
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Embodiment 1
[0041] Example 1 Establishment of an assessment model for recurrence and / or survival consequences or prognosis of renal cell carcinoma
[0042] By screening 80 patients with kidney cancer from Fudan University Cancer Hospital as a discovery set, the corresponding gene expression data were extracted by RNA-seq (transcriptome sequencing technology), and the Lasso Cox regression model was used to predict the biochemical recurrence of these patients. related genes. Finally, a genetic decision-making model based on 5 genes was determined and constructed (see figure 1 ). And according to the risk of the decision model, the following scoring formula was used to obtain the risk value, and the patients were divided into high-risk group and low-risk group of renal cancer.
[0043] Risk value=(1.6631×ASPA expression level)+(0.2026×TMEM38B expression level)+(0.1667×PTGR2 expression level)+(-2.3969×TXLNB expression level)+(-2.0618×ADGRE2 expression level).
[0044] The mRNA expression v...
Embodiment 2
[0046] Embodiment 2 is used for the multigene test kit of kidney cancer prognosis
[0047] The multigene kit used for renal cancer prognosis in this embodiment includes:
[0048] Total RNA extraction reagent Trizol;
[0049] Chloroform (chloroform);
[0050] isoamyl alcohol;
[0052] DEPC water (DD1005);
[0053] Ethylene glycol pyrophosphate (DEPC);
[0054] Anti-RNase solution (RNaseZap);
[0055] Reverse transcription kit;
[0056] iQ SYBR Green Supermix;
[0057] The polynucleotide primer whose sequence is SEQ ID NO: 1-10.
[0058] After using the kit to perform real-time quantitative PCR (qRT-PCR) detection of each gene, the initial data result is represented by the Ct value (cycle threshold), that is, the number of cycles required for the fluorescent signal in each reaction system to reach the set threshold. The Ct value of each sample has a linear relationship with the logarithm of the initial copy number of the sample, and the highe...
Embodiment 3
[0061] Preparation and pretreatment of embodiment 3 sample RNA
[0062] (1) For the tissue RNA extraction experiment, soak the tip and tweezers in DEPC water overnight, then sterilize under high temperature and high pressure, pre-cool the centrifuge at 4°C, and wipe the test bench, pipette gun, gloves, etc. with anti-RNase Zap.
[0063] (2) Tissue homogenate Take 50-100mg tissue sample, mince it slightly with a sterile scalpel blade, put it into a 1.5mL EP tube, add 500μL Trizol reagent, fully homogenize with an electric tissue grinder, and then add 500μL Trizol reagent .
[0064] (3) Add 0.2mL chloroform to every 1mL Trizol reagent homogenate sample, and tightly cap the EP tube. Shake vigorously for 15 seconds, and place at room temperature for 3 minutes. Centrifuge at 12,000 rpm at 4°C for 15 minutes (pre-cool the centrifuge). After centrifugation, the mixed system will be divided into a colorless aqueous phase in the upper layer, a protein in the middle layer and a red p...
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