A group of markers and their application in the differential diagnosis of benign and malignant thyroid follicular neoplasms
A technology of differential diagnosis and markers, applied in the field of biomedicine, can solve the problems of low sensitivity and specificity of benign and malignant judgment indicators, limited and lack of biological indicators of follicular tumors, and achieve the effect of avoiding overdiagnosis and treatment
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Embodiment 1
[0043] Example 1: Screening of methylation markers for benign and malignant thyroid tumors
[0044] The inventor collected paraffin wax specimens of thyroid follicular tumors diagnosed by the Department of Pathology of Peking Union Medical College Hospital between 2010 and 2016, a total of 128 cases (including 46 cases of FTA, 36 cases of TT-UMP and 46 cases of FTC), extracted DNA, and performed DNA analysis. Magnetic bead purification (Backman Ampure Xp beads); Methylation detection and analysis by bisulfite sequencing technology (RRBS), first look for differentially methylated molecules in the training set (consisting of 33 cases of FTA and 33 cases of FTC) .
[0045] Methylation detection and analysis was completed by Shanghai Kunyuan Biotechnology Co., Ltd.: take 100ng of purified sample DNA for enzyme digestion reaction, the reaction system is shown in Table 1, and the reaction temperature is as follows: 37°C for 3 hours, 80°C for 20 minutes, and 4°C for storage . Among...
Embodiment 2
[0053] Example 2 Using 70 differentially methylated markers to construct an assessment model for the potential of malignant thyroid cancer
[0054] After performing quality control analysis on the sequencing results of each sample in Example 1, the sequences of 70 marker segments were selected and calculated according to the calculation method of the methylation value corresponding to each marker (methylation value The calculation method refers to GuoS, et al. (2017) Identification of methylation haplotype blocks aids indeconvolution of heterogeneous tissue samples and tumor tissue-of-originmapping from plasma DNA. Nat Genet 49(4):635-642.), if there is a section If the sequencing depth of a certain CpG site is lower than 10×, the methylation value of this segment is NA. A sample-marker numerical matrix is formed, and some exemplary data are shown in Table 3. The construction of the model was completed by Shanghai Kunyuan Biotechnology Co., Ltd.
[0055] Table 3 Marker met...
Embodiment 3
[0073] Example 3 Methylation Markers Used in the Differentiation of Benign and Malignant Thyroid Tumors
[0074] In this example, the model constructed in Example 2 was used to differentiate benign from malignant thyroid follicular tumor samples from 26 cases. The process is as follows:
[0075] According to the method of Example 2, a sample-marker numerical matrix of 26 samples similar to Table 3 was constructed.
[0076] Open the R program package and import the numerical matrix of 26 samples-markers to be evaluated
[0077] valdata=read.delim("storage path of sample-marker matrix", sep="\t", as.is=T, row.names=1, header=T, check.names=F)
[0078] Transpose the matrix into a matrix with row names of sample names and columns of methylation marker names
[0079] input=t(valdata)
[0080] And use the adjacent value complement method to make up the NA value
[0081] library(DMwR)
[0082] imputed = knnImputation(imput)
[0083] Reimport the established assessment model of...
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