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Primer group for detecting ENG gene mutation, purpose of primer group and kit containing primer group

An ENG gene and primer set technology, applied in the field of primer sets, can solve the problems of inability to find mutation sites, false positives, and large limitations, and achieve the effects of high accuracy, simple method, and ensuring accuracy and uniqueness.

Inactive Publication Date: 2016-08-24
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, fluorescent quantitative PCR is commonly used for detection, but the method for detecting mutations by this method is too limited. It can only detect known mutation points, and cannot find unknown mutation sites, and there is a risk of false positives.

Method used

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  • Primer group for detecting ENG gene mutation, purpose of primer group and kit containing primer group
  • Primer group for detecting ENG gene mutation, purpose of primer group and kit containing primer group
  • Primer group for detecting ENG gene mutation, purpose of primer group and kit containing primer group

Examples

Experimental program
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Effect test

Embodiment 1

[0031] According to the 1-15 exon regions, 3'UTR and promoter sequences of ENG published by NCBI Gene Bank (NC-000012), a primer set for detecting whether the ENG gene is mutated was designed, and the primer set included the above The PCR amplification primer set and PCR sequencing primer set of the region, the results are shown in Table 1 and Table 2.

Embodiment 2

[0033] A kit for detecting ENG gene mutations, comprising the following components:

[0034] There are 19 pairs of PCR amplification primers and PCR sequencing primers corresponding to the 1st to 15th exon regions, 3'UTR and promoter sequences of the ENG gene, each primer is 1OD, and the primer sequences of each pair are shown in Table 1. and Table 2.

[0035] This kit is stored at -20°C, and repeated freezing and thawing should be avoided as much as possible.

Embodiment 3

[0037] A kit for detecting ENG gene mutations, comprising the following components:

[0038](1) There are 19 pairs of PCR amplification primers and PCR sequencing primers corresponding to the 1st to 15th exon regions, 3'UTR and promoter sequences of the ENG gene, and each primer is 1OD. For the primer sequences of each pair, see Table 1 and Table 2.

[0039] (2) PCR amplification reagent: 2.5mM dNTP mixture 40μl, 5×amplification buffer (containing Mg 2+ ) 100μl, 5Units / ul Taq DNA polymerase 5μl.

[0040] (3) PCR product purification reagents: 1 U / ul SAP enzyme 20 μl, 10 U / ul ExoI enzyme 10 μl.

[0041] (4) PCR sequencing reagents: BigDye 3.1 mix 50μl, EDTA solution (0.125M) 50μl, absolute ethanol 1ml, ethanol solution (75%) 1.5mL, HIDI solution 500μl.

[0042] This kit is stored at -20°C, and repeated freezing and thawing should be avoided as much as possible.

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Abstract

The invention discloses a primer group for detecting ENG gene mutation. The primer group comprises an ENG gene first to fifteenth exon regions, a PCR (polymerase chain reaction) amplification primer group corresponding to 3'UTR and a starter and a PCR sequence testing primer. The invention also discloses a purpose of the primer group and a kit containing the primer group. The primer group can be used for specifically identifying whether the ENG gene mutation occurs or not; the accuracy and the uniqueness of the detection result are ensured.

Description

technical field [0001] The present invention relates to a primer set, in particular to a primer set for detecting ENG gene mutation, and also relates to the use of the primer set and the kit it constitutes. Background technique [0002] ENG (endoglin, also known as CD105) is located at 9q34 and is a cell membrane glycoprotein that participates in vascular development and remodeling by regulating the response of cell transformation factor β. Mutations in it can cause hereditary hemorrhagic telangiectasia (HHT) or hereditary pulmonary arterial hypertension (PAH). Some mutation carriers have both conditions at the same time. [0003] HHT is an autosomal dominant genetic hemorrhagic disorder characterized by multiple telangiectasias of the skin and mucous membranes accompanied by recurrent hemorrhage. Among them, HHT1 is caused by ENG gene mutations. Both HHT1 and HHT2 patients have epistaxis, telangiectasia, gastrointestinal bleeding, migraine and so on. However, compared wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 卢文菊张晨婷王健
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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