Pulmonary arterial hypertension associated mutant BMP9 gene and application thereof

[0093] research population

[0094] Patient group: Patients with pulmonary arterial hypertension were selected from Shanghai Pulmonary Hospital Affiliated to Tongji University and Fuwai Hospital of Chinese Academy of Medical Sciences. All patients underwent right heart catheterization. The diagnostic criteria for PAH were: mean pulmonary artery pressure (mPAP) ≥ 25 mmHg and pulmonary capillary wedge pressure (PCWP) ≤ 15 mmHg measured by right heart catheterization at sea level at rest. After excluding all known secondary factors causing PAH, the patient was diagnosed with idiopathic PAH. Finally, 269 IPAH patients were selected.

[0095] Control group 1: 1884 cases of healthy controls were selected from Beijing Novogene Technology Co., Ltd. Both the case group and the control group were of Chinese Han nationality and had no blood relationship. Genomic DNA was extracted from peripheral blood leukocytes of patients and controls.

[0096] Control group 2: The whole exome sequenc

[0135] The 16 IPAH patients carrying 16 kinds of BMP9 gene variations in the detection example 1 carry the BMP9 genome sequence in the blood, BMP9 has 2 exons, the exon sequence can be amplified by PCR method, and the amplified product can be detected by Sanger sequencing method sequence.

[0136] 1 Reagent:

[0137] (1) Human Blood Genomic DNA Purification Kit DP318-02 (Tiangen Biochemical Technology, Beijing, China);

[0138] (2) 10% SDS: Dissolve 10.0g SDS in 90ml water, adjust pH to 7.2, add water to make up to 100ml;

[0139] (3) TE: 10mM Tris-HCl (pH 8.0), 1mM EDTA-Na2 (pH 8.0);

[0140] (4) 50X TAE electrophoresis buffer: 242.0g Tris-HCl, 27.5ml glacial acetic acid, 100ml 0.5mM EDTA-Na2, final volume 1 liter;

[0141] (5) All restriction endonucleases were purchased from New England Biolabs, USA;

[0142] (6) The primers used were purchased from Beijing Aoke Biological Company.

[0143] 2 Instruments and equipment:

[0144] (1) PCR instrument: DNA engine American Bio