Detection method for simultaneous implementation of PGT-A (Preimplantation Genetic Testing Aneuploidy) and PGT-M (Preimplantation Genetic Testing for Monogenic/Single Gene Defects) and kit of detection method

A technology of PGT-A and detection kits, which is applied in the direction of biochemical equipment and methods, microbiological measurement/inspection, etc., can solve the problem of high single-cell template allele drop-out rate, achieve improved probability, simple operation, The effect of simple method

Pending Publication Date: 2019-09-27
阿吉安(福州)基因医学检验实验室有限公司
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AI Technical Summary

Problems solved by technology

[0007] In order to overcome the above-mentioned defects in the prior art, the technical problem to be solved by the present invention is to provide a fast and convenient method for simultaneously performing PGT-A and PGT-M to solve the problem of high single-cell template allele dropout rate. Detection kits and detection methods thereof

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  • Detection method for simultaneous implementation of PGT-A (Preimplantation Genetic Testing Aneuploidy) and PGT-M (Preimplantation Genetic Testing for Monogenic/Single Gene Defects) and kit of detection method
  • Detection method for simultaneous implementation of PGT-A (Preimplantation Genetic Testing Aneuploidy) and PGT-M (Preimplantation Genetic Testing for Monogenic/Single Gene Defects) and kit of detection method
  • Detection method for simultaneous implementation of PGT-A (Preimplantation Genetic Testing Aneuploidy) and PGT-M (Preimplantation Genetic Testing for Monogenic/Single Gene Defects) and kit of detection method

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Effect test

Embodiment 1

[0052] Simultaneous detection kit for PGT-A and PGT-M, including cell lysate, pre-amplification mixture, amplification mixture, end repair reaction solution, end repair enzyme, DNA ligase buffer, DNA ligase, linker , enzyme-free sterile water, STR primers, 10×PCR buffer, dNTP mixture, Taq polymerase, restriction endonuclease and restriction endonuclease.

Embodiment 2

[0054] Detection of PGT-A / PGT-M in preimplantation embryos of patients with monogenic homocystinuria with methylenetetrahydrofolate reductase deficiency;

[0055] 1. Single-cell genome-wide amplification of blastocyst trophoblast cells

[0056] 1) Sample processing

[0057] Mark the last four digits of the number, the date, and the serial number of the experimental sample on the cap of the 0.2ml PCR tube; centrifuge the sample at 200×g for 3 minutes.

[0058] 2) Cell Lysis

[0059] The cell lysate was briefly centrifuged for later use, and divided into n PCR reaction tubes according to 0.6 μL of cell lysate per person (each sample), and the cell lysate was added to the PCR tube containing the processed sample and cell lysis buffer, Run the following program according to Table 1 to obtain the cleavage reaction product.

[0060] Table 1

[0061]

[0062] 3) Pre-amplification

[0063] According to the number n of samples, take n parts of the pre-amplification mixture and ...

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Abstract

The invention discloses a detection method for simultaneous implementation of PGT-A (Preimplantation Genetic Testing Aneuploidy) and PGT-M (Preimplantation Genetic Testing for Monogenic / Single Gene Defects) and a kit of the detection method and belongs to the technical field of biological detection. The method comprises the following steps: acquiring blastaea trophoblast cells, and carrying out single cell whole genome amplification so as to obtain an amplification product; designing an STR (Simple Tandem Repeat) primer according to the family system of a single-gene patient; carrying out second-generation sequencing on the amplification product; carrying out STR linkage analysis, restrictive fragment length polymorphic analysis and fragmentation analysis on the amplification product and the STR primer. The invention further provides a detection kit for implementing PGT-A and PGT-M. By adopting the detection method and the kit thereof, STR sites which are closely associated with disease causing genes are selected for haplotype analysis, problems that a conventional method has limit on the number of single cell templates and has a high allelic gene releasing rate can be solved, the detection method and the kit can be applied to chromosome aneuploidy detection and single-gene inheritance disease detection before implantation of test tube babies, and are low in detection cost and convenient in operation.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a detection method for simultaneously performing PGT-A and PGT-M and a kit thereof. Background technique [0002] According to the "China Infertility Status Survey Report" issued by the China Population Association and the National Family Planning Commission, the incidence of infertility in China is 12.5-15%, and the number of patients exceeds 40 million. 1 pair had infertility problems. After the opening of the two-child policy, there are about 90 million couples in China who meet the second-child standard, which has led to a sharp increase in the proportion of elderly women among the pregnant population. Infertile women older than 35 years old, patients with repeated spontaneous abortions, and patients with repeated embryo implantation failures face more difficult fertility problems. [0003] According to statistics, the incidence rate of genetic dise...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6806
CPCC12Q1/6883C12Q1/6806C12Q2600/156
Inventor段佳丽郭文浒郑邦烈薛怡婷陈浩
Owner阿吉安(福州)基因医学检验实验室有限公司