SNP molecular marker of RANTES gene related to generation of preterm birth and kit for detecting RANTES gene

A molecular marker and kit technology, applied in the field of molecular biology, to achieve the effect of good clinical application value

Inactive Publication Date: 2015-05-20
THE MILITARY GENERAL HOSPITAL OF BEIJING PLA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the role of RANTES gene polymorphisms in t

Method used

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  • SNP molecular marker of RANTES gene related to generation of preterm birth and kit for detecting RANTES gene
  • SNP molecular marker of RANTES gene related to generation of preterm birth and kit for detecting RANTES gene
  • SNP molecular marker of RANTES gene related to generation of preterm birth and kit for detecting RANTES gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Establishment of the polymorphic site IN1.1T / C method for detecting the RANTES gene

[0027] 1. Design and synthesis of primers

[0028] The sequence of the specific primer pair for detecting the genotype of the RANTES gene IN1.1T / C site is:

[0029] Upstream primer: 5'-CCTGGTCTTGACCACCACA-3',

[0030] Downstream primer: 5'-GCTGACAGGCATGAGTCAGA-3'.

[0031] 2. Genomic DNA extraction

[0032] 3. Establishment of IN1.1T / C method for detecting polymorphic sites of RANTES gene

[0033] (1) Extract the genomic DNA of the sample to be tested, use it as a template, and use the above primer pair as amplification primers to perform a PCR reaction; the PCR reaction is pre-denatured at 95°C for 5 minutes and then subjected to two stages of amplification: the first stage is Denaturation at 94°C for 30s, annealing temperature decreased by 0.5°C for 30s per cycle from 58°C, extension at 72°C for 50s, a total of 16 cycles; the second stage was denaturation at 94°C for 30...

Embodiment 2

[0035] Example 2 Analysis of polymorphic site IN1.1T / C of RANTES gene and susceptibility to preterm birth

[0036] Using the method established in Example 1, the IN1.1T / C site genotype of the RANTES gene in the Chinese Han population (569 cases of premature infants and 673 cases of full-term infants) was analyzed. 1. Test object

[0037] The subjects of the study were neonates admitted to Bayi Children's Hospital affiliated to Beijing Military Region General Hospital from February 2009 to May 2011. All subjects were blood-unrelated Han Chinese residing in and around Beijing. Premature babies are spontaneously premature. According to the gestational age at birth, they were divided into 3 groups: moderately premature (33-36 gestational weeks); preterm (<32 gestational weeks); extremely premature (<28 gestational weeks). Normal controls were randomly selected singleton full-term infants born to pregnant women with no history of premature rupture of membranes and preterm birth....

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Abstract

The invention provides a SNP molecular marker of RANTES gene related to generation of preterm birth and a kit for detecting the RANTES gene. According to the invention, statistic analysis of a large number of clinical samples discovers that polymorphic site In1.1T/C of RANTES gene is related to generation of preterm birth. The invention also provides the kit for detecting the polymorphic site In1.1T/C of the RANTES gene. Based on a PCR method, a pair of primers are designed, nucleotide sequence is shown in SEQ ID NO.1-2, PCR amplification is carried out on to-be-detected sample genome DNA, and direct sequencing is carried out on the amplified product to be used for determining genotype of the polymorphic site In1.1T/C of the RANTES gene. The kit is simple to operate, high in sensitivity, strong in accuracy and low in detection cost and has important application value.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a detection kit for detecting the SNP molecular marker of the RANTES gene related to the occurrence of premature birth and the molecular marker. Background technique [0002] Preterm birth (Preterm Birth, PTB) is an important cause of neonatal morbidity and death, and it is also the cause of complications and sequelae in infants and young children such as cerebral palsy, developmental delay, retinopathy of prematurity, chronic lung disease, and hearing and visual impairment. Key factor. 75% of neonatal morbidity and death are related to premature birth. About 15 million premature babies are born worldwide each year, and this number is rising. According to the data released by the World Health Organization, the incidence rate of premature babies in my country was 7.8% in 2011, and nearly 1.1 million premature babies were born, the number of births is second only to Ind...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 王艳封志纯张小爱杨晓
Owner THE MILITARY GENERAL HOSPITAL OF BEIJING PLA
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