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Primer group for detecting ALK-1 (activin receptor type II-like kinase-1) gene mutation, and application thereof as well as kit comprising primer group

An ALK-1, primer set technology, applied in the direction of DNA/RNA fragment, recombinant DNA technology, microbial determination/inspection, etc., can solve dangerous problems and achieve the effect of ensuring accuracy and uniqueness, and the method is simple

Active Publication Date: 2014-04-02
广州瑞能精准医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, patients with pulmonary arterial hypertension with ALK1 gene mutation are younger than patients with bone morphogenic protein 2 (BMPR2) gene mutation, and once the disease progresses more rapidly, it is more dangerous

Method used

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  • Primer group for detecting ALK-1 (activin receptor type II-like kinase-1) gene mutation, and application thereof as well as kit comprising primer group
  • Primer group for detecting ALK-1 (activin receptor type II-like kinase-1) gene mutation, and application thereof as well as kit comprising primer group
  • Primer group for detecting ALK-1 (activin receptor type II-like kinase-1) gene mutation, and application thereof as well as kit comprising primer group

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Effect test

Embodiment 1

[0046] Based on the 1-10 exon regions, 3'UTR and promoter sequence of ALK-1 published by NCBI Gene Bank (NC-000012), a primer set for detecting ALK-1 gene mutation was designed. The set includes the PCR amplification primer set and PCR sequencing primer set of the above regions. The results are shown in Table 1 and Table 2.

Embodiment 2

[0048] A kit for detecting ALK-1 gene mutation, including the following components:

[0049] The PCR amplification primer set and PCR sequencing primer set corresponding to the 1st to 10th exon region, 3'UTR and promoter sequence of ALK-1 gene, a total of 15 pairs, each primer 1OD, each pair of primer sequences see Table 1 and Table 2.

[0050] This kit is stored at -20°C to minimize repeated freezing and thawing.

Embodiment 3

[0052] A kit for detecting ALK-1 gene mutation, including the following components:

[0053] (1) PCR amplification primer set and PCR sequencing primer set corresponding to the 1st to 10th exon regions, 3'UTR and promoter sequence of ALK-1 gene, a total of 15 pairs, each primer 1OD, each pair of primers See Table 1 and Table 2 for the sequence.

[0054] (2) PCR amplification reagents: 2.5mM dNTP mixture 40μl, 5× amplification buffer (containing Mg 2+ ) 100μl, 5Units / ul Taq DNA polymerase 5μl.

[0055] (3) PCR product purification reagents: 1U / ul SAP enzyme 20μl, 10U / ul ExoI enzyme 10μl.

[0056] (4) PCR sequencing reagents: BigDye3.1mix50μl, EDTA solution (0.125M) 50μl, absolute ethanol 1ml, ethanol solution (75%) 1.5mL, HIDI solution 500μl.

[0057] This kit is stored at -20°C to minimize repeated freezing and thawing.

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Abstract

The invention discloses a primer group for detecting ALK-1 (activin receptor type II-like kinase-1) gene mutation, at least comprising ALK-1 gene first to tenth exon regions, and a PCR (Polymerase Chain Reaction) amplification primer group and a PCR sequencing primer group which correspond to any region of a 3' UTR (Untranslated Region) and a promoter. The invention further discloses an application of the primer group, a kit comprising the primer group, and a method for carrying out PCR amplification on an ALK-1 gene by the primer group. The primer group can be used for specially identifying whether the ALK-1 gene is mutated or not so as to ensure the accuracy and the uniqueness of a detected result.

Description

Technical field [0001] The present invention relates to primers, in particular to a primer set for detecting ALK-1 gene mutations. The present invention also relates to the use of the primer set and a kit containing the primer set. Background technique [0002] Activin receptor type Ⅱ-like kinase-1 (ALK-1, also known as ACVRL1) is located at 12q11-q14, contains 10 exons, and the genome is about 15kb+ in length. Its mutation can cause hereditary hemorrhagic telangiectasia HHT or hereditary pulmonary arterial hypertension (PAH). Some mutation carriers have both symptoms. HHT is an autosomal dominant hereditary hemorrhagic disease, characterized by multiple telangiectasias in the skin and mucous membranes with repeated bleeding. It can generally be divided into two phenotypes, type I HHT and type II HHT. Among them, type Ⅱ HHT is caused by ALK-1 gene mutation. The most common clinical manifestation of HHT-Ⅱ is recurrent spontaneous bleeding or mild traumatic bleeding, with nose bl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 卢文菊张晨婷王健
Owner 广州瑞能精准医学科技有限公司
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