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Method for identifying SNP genotype-based monozygotic twins, fraternal twins and second polar body participated fertilized twins

A technology for fraternal twins and identification methods, which is applied in biochemical equipment and methods, and microbial measurement/inspection, etc., can solve the problems of inability to accurately determine twins, inability to identify genetic differences, and high similarity of STR genetic markers, achieving sensitivity and High accuracy, simple operation and high analysis efficiency

Pending Publication Date: 2020-08-14
THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this case, it is impossible to effectively identify the genetic differences between the egg cell and the second polar body; secondly, if both parents come from the same place, affected by genetic drift, the degree of similarity of their STR genetic markers may be very small. High, making it impossible to accurately determine whether twins are fraternal twins
[0009] At present, there is no identification method for monozygotic twins, fraternal twins and second polar body involved in the fertilization of twins based on the SNP genotype of this application

Method used

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  • Method for identifying SNP genotype-based monozygotic twins, fraternal twins and second polar body participated fertilized twins
  • Method for identifying SNP genotype-based monozygotic twins, fraternal twins and second polar body participated fertilized twins
  • Method for identifying SNP genotype-based monozygotic twins, fraternal twins and second polar body participated fertilized twins

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Embodiment 1

[0050] This embodiment provides a method for identifying monozygotic twins, fraternal twins and second polar body twins involved in fertilization based on the SNP genotype, comprising the following steps:

[0051] S1. Extract the genomic DNA of the twins to be tested and their parents, measure the DNA concentration, and uniformly standardize;

[0052] S2. Add an appropriate amount of NaOH to the sample to denature the DNA into a single strand, add the whole genome amplification reagent after neutralization, and incubate overnight at a constant temperature of 37°C;

[0053] S3. The amplified product undergoes a controlled enzymatic treatment that does not require gel electrophoresis to become fragmented DNA;

[0054] S4. Precipitating the DNA fragments by adding isopropanol, and enriching the fragmented DNA by centrifugation at 4° C., thereby being purified;

[0055] S5. After the precipitated DNA is dried in the air, a hybridization buffer reagent is added to redissolve the D...

Embodiment 2

[0068] This embodiment provides an application example of the method of the present invention.

[0069] In 2019, a pair of twins were born in the Obstetrics and Gynecology Hospital Affiliated to Fudan University. The twins passed B-ultrasound examination before delivery and were found to be monochorionic twins. According to the traditional view, they are identical twins derived from the division of the same fertilized egg. But after the birth, medical staff discovered that the twins were not identical. According to the method provided by the present invention, the identification of monozygotic twins, fraternal twins and twins whose second polar body participates in fertilization is carried out.

[0070] Step 1, DNA extraction:

[0071] Using a commercially available DNA extraction kit (Genomic DNA Small Extraction Kit, Shanghai Laifeng Biotechnology Co., Ltd.), the genomic DNA of the twins and their parents was extracted.

[0072] Step 2, STR genetic marker verification: us...

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Abstract

The invention relates to a method for identifying SNP genotype-based monozygotic twins, fraternal twins and second polar body participated fertilized twins. The method comprises the following steps: analyzing the SNP genotype similarity level of father being homozygote and mother being heterozygote in twins, the twins are determined to be fraternal twins if the similarity level is about 50 percent, otherwise, the twins are determined to be non-fraternal twins if the similarity level is more than 90 percent. If the twins are determined to be non-fraternal twins, further analysis is required: the SNP genotype is completely the same in the twins, the twins are determined to be monozygotic twins; and otherwise, if the similarity level x is more than 90 percent and less than 100 percent, the twins are determined to be second polar body participated fertilized twins. The novel and smart identification method is provided for the first time, the blank for identifying monozygotic, fraternal andsecond polar body participated fertilized twins in medical and expert testimony, has high sensitivity and high accuracy, is easy in sample acquisition, simple in operation and high in analyzing efficiency, and is widely applicable to the fields of expert testimony / medical identification and the like.

Description

technical field [0001] The invention relates to the technical field of human genetics, in particular to a method for identifying monozygotic twins, fraternal twins and twins whose second polar body participates in fertilization based on SNP genotype. Background technique [0002] Twins are common, and the traditional classification method divides them into identical twins and fraternal twins. There is no controversy about the classification of fraternal twins at present, that is, two eggs are released in the same menstrual cycle, and they are fertilized with different sperm and develop into two twins with their own chorion, amniotic cavity and placenta, which are the same as non-twin brothers and sisters ~50 % Independent individuals of the same genetic material. The traditional division mechanism of monozygotic twin formation has been challenged in recent years: cases of "semi-identical twins" (identical genes from the mother and partially overlapping genes from the father...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/156
Inventor 王红艳史植文李笑天李成涛王一
Owner THE OBSTETRICS & GYNECOLOGY HOSPITAL OF FUDAN UNIV
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