Kit for predicting thyroid tumors to be benign or malignant
A technology of thyroid tumors and kits, applied in the biological field, can solve the problem of low accuracy in predicting benign and malignant, and achieve the effects of strong clinical applicability, wide applicability, and low cost
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Embodiment 1
[0022] Using bioinformatics methods, based on machine learning theory, through the screening and analysis of benign and malignant thyroid tumor gene expression microarray independent data sets in the public database (Gene Expression Omnibus, GEO), using iterative Bayesian model averaging (iterativeBMA) ) screened a small number of gene combinations, and verified them with three other independent data sets in the GEO database, and finally established a real-time fluorescent quantitative PCR (QPCR) method to detect a small number of gene combinations, and achieved the prediction of benign and malignant thyroid tumors through the Bayesian average model. Purpose.
[0023] (1) Two-step method for screening tumor marker gene combinations: linear model (limma) of microarray data + iterative Bayesian model averaging method;
[0024] Firstly, the linear model (limma) for microarray data was used, and GSE29315 in the GEO public database was used as the training set to screen 71 differen...
Embodiment 2
[0046] 1.1 Specimen collection
[0047] A total of 70 patients with thyroid nodules diagnosed by outpatient examinations in Shanghai Renji Hospital from 2012 to 2013 were collected, including 48 females (age 20-67 years old) and 22 males (age 23-74 years old). After thyroidectomy in these patients, some samples were used for HE staining for clinicopathological diagnosis (gold standard), and the other part was kept in RNA preservation solution RNAlater (Invitrogen, USA) and stored at -80°C. Diagnosed by the clinical pathology department, there were 39 cases of benign thyroid tumors (20 cases of follicular adenoma, 11 cases of Hashimoto's thyroiditis, 8 cases of nodular thyroid hyperplasia) and 31 cases of malignant thyroid tumors (29 cases of papillary thyroid hyperplasia) among the 70 samples. Thyroid carcinoma, 1 case of follicular thyroid carcinoma, 1 case of anaplastic carcinoma). RNA was extracted using the TRIZOL method (Invitrogen, U.S.), and carried out according to th...
Embodiment 3
[0129] Example 3 Fluorescent quantitative PCR kit for predicting benign and malignant thyroid tumors
[0130] Prepare a fluorescent quantitative PCR kit for predicting benign and malignant thyroid tumors, which contains:
[0131]
[0132] 100 people were randomly selected to form the test group, including patients who were unknown whether they had malignant thyroid tumors, and patients who were unknown whether they had benign thyroid tumors.
[0133] Under the guidance of ultrasound, fine-needle aspiration samples of thyroid nodules from the subjects to be tested were collected and stored in RNA preservation solution RNAlater at -80°C. RNA was extracted by TRIZOL method.
[0134] 1.1 Extract RNA as in Example 2 using the TRIZOL method.
[0135] 1.2 Reverse transcription PCR (RT–PCR)
[0136] Prepare RT reaction solution on ice, using 25 μl system.
[0137] 25μl system (all units are μl)
[0138]
[0139] 75°C for 5min, anneal on ice for 5min, centrifuge for 2-3s
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