The application of braf gene detection material in the preparation of acth type pituitary adenoma molecular pathological diagnosis and typing products
A pituitary adenoma, gene technology, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc.
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Embodiment 1
[0046] Example 1. Whether the BRAF gene is mutated or not, and its application in assisting in the identification of subtypes of ACTH-type pituitary adenomas
9 example A
[0047] Whole-exome sequencing was performed on the tumors of 9 cases of ACTH-type pituitary adenomas and the DNA of their own blood samples (the sequencer was Illumina Hiseq 2500). The results showed that BRAF gene mutations appeared in 2 cases of tumors, and the tumor volume was small, and the other 7 cases The BRAF gene in the tumor is wild type, and the tumor volume is large; therefore, ACTH pituitary adenomas are divided into two subtypes: ACTH pituitary adenoma wild type and ACTH pituitary adenoma mutant.
[0048]BRAF genotype detection was performed in another 149 cases of ACTH-type pituitary adenomas (from Huashan Hospital Affiliated to Fudan University, all patients gave informed consent) after 9 samples were verified by Sanger to exclude false positives. The detection method of a total of 149 samples details as follows:
[0049] First, according to the BRAF gene (the nucleotide sequence is the sequence 1 in the sequence table), the primer pairs for amplifying it are d...
Embodiment 2
[0072] Example 2. Application of whether the BRAF gene is mutated or not in the auxiliary identification of ACTH-type pituitary adenoma subtypes
[0073] 150 cases of non-ACTH type pituitary adenoma patients (comprising 50 cases of hormone-free type, prolactin type, growth hormone type) tumors and 149 cases of ACTH type pituitary adenoma patients of embodiment 1 (derived from Fudan Huashan Hospital Affiliated to the University, and all patients gave informed consent) The tumors were tested for BRAF gene mutation, and the specific methods were as follows:
[0074] Genomic DNA of the tumors of each pituitary adenoma patient was extracted, and PCR amplification was performed with primer pair 1 and primer pair 2 of Example 1 to obtain PCR amplification products, which were then sequenced.
[0075] The sequencing results are as follows: the nucleotide sequences of the PCR amplification products of the tumors of 150 patients with non-ACTH-type pituitary adenomas were all consistent ...
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