Application of BRAF gene detector to preparation of ACTH type pituitary adenoma molecular pathological diagnosis and parting product

A pituitary adenoma, gene technology, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc.

Active Publication Date: 2016-04-20
SHANGHAI JIAO TONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are currently no specific molecular diagnostic markers to differentiate them

Method used

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  • Application of BRAF gene detector to preparation of ACTH type pituitary adenoma molecular pathological diagnosis and parting product
  • Application of BRAF gene detector to preparation of ACTH type pituitary adenoma molecular pathological diagnosis and parting product
  • Application of BRAF gene detector to preparation of ACTH type pituitary adenoma molecular pathological diagnosis and parting product

Examples

Experimental program
Comparison scheme
Effect test

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 IlluminaHiseq2500). The results showed that BRAF gene mutations appeared in 2 tumors, and the tumor volume was small. Therefore, ACTH pituitary adenoma is 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 designed, a total of 2 pairs of primer pairs, each pair of primer pairs can amplify a partial fragment of th...

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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Abstract

The invention discloses application of a BRAF gene detector to preparation of an ACTH type pituitary adenoma molecular pathological diagnosis and parting product. It is proved through tests that BRAF gene mutation is peculiar to ACTH type pituitary adenoma and not found in other types of pituitary adenoma, so that the BRAF gene mutation can serve as a specific molecular pathological diagnosis marker of the ACTH type pituitary adenoma. According to the result that a BRAF gene is mutant or not, the ACTH pituitary adenoma can be further divided into two subtypes, namely the BRAF mutant type and the wild type, the mean volume of the BRAF mutant type ACTH pituitary adenoma is small, the attack degree is low, and the midnight serum cortisol concentration is less than or equal to 22 ug / dl; the mean volume of the wild type ACTH pituitary adenoma is large, the attack degree is high, and the midnight serum cortisol concentration is greater than 22 ug / dl. Therefore, the different subtypes of the ACTH pituitary adenoma can be judged by detecting whether the BRAF gene is mutant or not, and different clinical treatment strategies are adopted in time accordingly.

Description

technical field [0001] The invention relates to the application of a BRAF gene detection substance in the preparation of ACTH type pituitary adenoma molecular pathological diagnosis and typing products, and belongs to the field of biotechnology. Background technique [0002] Pituitary adenomas are the most common neuroendocrine tumors in the human body and one of the most common brain tumors (the incidence rate is the second). According to its different endocrine characteristics, it can be divided into several subtypes such as prolactin type, growth hormone type, adrenocorticotropic hormone type, thyroid stimulating hormone type, hormone-free type and mixed type. Tumors secrete various hormones abnormally, leading to significant endocrine disorder symptoms such as infertility, acromegaly, Cushing syndrome, and hyperthyroidism. Pituitary tumors are insidious in onset and slow in growth. When patients present with clinical symptoms and go to see a doctor, the tumor is often h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/156
Inventor 师咏勇赵曜黄传新陈剑华宋智健马增翼张启麟间学敏李士其王镛斐叶红英叶钊
Owner SHANGHAI JIAO TONG UNIV
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