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Specific primer for detecting gene single nucleotide polymorphism site of premature delivery-related beta-2-adrenergic receptor

A single nucleotide polymorphism and receptor gene technology, applied in the field of molecular biology, can solve the problems such as no public report on the detection mutation site rs1042713, and achieve the effects of easy promotion, high specificity and low cost

Active Publication Date: 2015-09-23
FIRST PEOPLES HOSPITAL OF YUNNAN PROVINCE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] After literature search, there is no public report on detecting the mutation site rs1042713 with the same allele-specific PCR method as in the present invention

Method used

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  • Specific primer for detecting gene single nucleotide polymorphism site of premature delivery-related beta-2-adrenergic receptor
  • Specific primer for detecting gene single nucleotide polymorphism site of premature delivery-related beta-2-adrenergic receptor
  • Specific primer for detecting gene single nucleotide polymorphism site of premature delivery-related beta-2-adrenergic receptor

Examples

Experimental program
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Embodiment 1

[0027] Example 1: Collect clinical blood samples and perform genomic DNA extraction

[0028] (1) Collect the peripheral blood of 20 clinically premature pregnant women and 20 full-term pregnant women each. All personnel will record the basic clinical data and testing data in detail. After signing the informed consent, each person will collect 3-5 ml of blood samples.

[0029] (2) Sample genomic DNA extraction

[0030] Genomic DNA was extracted from whole blood samples using a blood genomic DNA extraction kit.

[0031] Steps:

[0032] (1) Handling blood material

[0033] Take 200 μl of blood sample in a 1.5ml centrifuge tube.

[0034] (2) Add 20 μl Proteinase K solution and mix well.

[0035] (3) Add 200 μl buffer GB, mix thoroughly by inverting, place at 56°C for 10 min, and invert and mix several times during this time, the solution should become clear.

[0036] (4) Add 200 μl of absolute ethanol and mix well by inverting, at this time flocculent precipitation may appe...

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Abstract

The invention relates to a specific primer for detecting a gene single nucleotide polymorphism (SNP) site rs1042713 of a premature delivery-related beta-2-adrenergic receptor (ADRB2), and belongs to the technical field of molecular biology. Based on a site-specific PCR (polymerase chain reaction) principle, two upstream primers with tail ends 3minute paired with bases of two allelic genes with SNP respectively are designed, mismatched base (base A is mutated into base T) is introduced into 4th base at the ends 3minute of the specific primers of the two allelic genes to improve the availability of the primers, and a shared primer is positioned at the downstream part. A PCR amplification product is detected through agarose electrophoresis, and whether a sample contains the SNP site can be determined according to whether electrophoretic bands exist. In addition, the OD (optical density) value of the PCR product also can be measured through an enzyme-labeling instrument, whether the sample contains the SNP site is analyzed through comparison, and the method has relatively high sensitivity and is more suitable for large sample research. The specific PCR of the allelic genes is a simple, convenient, quick, reliable and low-cost SNP detection method and can be effectively applied to polymorphism screening of ADRB2, and the specific primer is of great significance in predicting and preventing premature delivery.

Description

technical field [0001] The invention relates to a method for detecting the β2 adrenoceptor gene ( beta-2-adrenergic receptor, ADRB2 ) The specific primer of single nucleotide polymorphism site rs1042713 belongs to the technical field of molecular biology. Background technique [0002] Preterm birth is one of the important causes of perinatal death, with an incidence rate of 5% to 15%, and the incidence rate of premature birth in my country is 7.8%. About 13 million women worldwide are born prematurely, which significantly increases neonatal mortality and morbidity, and has long-term adverse health effects. Premature babies are more likely to suffer from cerebral palsy, sensory impairment, learning disabilities and respiratory diseases than full-term babies. A large number of epidemiological surveys have found that premature birth is still an important factor for metabolic diseases such as cardiovascular disease and diabetes. According to the World Health Organization, 130 ...

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 董旭东刘锐宋玉竹陈晓
Owner FIRST PEOPLES HOSPITAL OF YUNNAN PROVINCE