Detection kit for chromosome abnormality before embryo implantation

A chromosomal abnormality and preimplantation technology, applied in the preimplantation chromosomal abnormality detection kit and its detection field, can solve problems such as low efficiency and complicated operation, achieve wide coverage and reduce false positive and false negative rates , to ensure the effect of reliability

Pending Publication Date: 2017-10-20
SUZHOU BASECARE MEDICAL DEVICE CO LTD
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  • Abstract
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Problems solved by technology

[0006] The invention discloses a pre-implantation chromosomal abnormality detection

Method used

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  • Detection kit for chromosome abnormality before embryo implantation
  • Detection kit for chromosome abnormality before embryo implantation
  • Detection kit for chromosome abnormality before embryo implantation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Preparation of Preimplantation Chromosomal Aneuploidy Detection Kit and Its Use

[0065] 1. Prepare a kit including the following components

[0066] 1.1 Reagents for single cell expansion: 1 tube of cell extraction buffer (310 μl / tube), 1 tube of cell lysis buffer (310 μl / tube), 1 tube of cell lysing enzyme (20 μl / tube), pre-amplification buffer (300 μl / tube) tube) 1 tube, pre-amplification enzyme (20μl / tube) 1 tube, amplification buffer (1.5ml / tube) 1 tube, amplification enzyme (60μl / tube) 1 tube.

[0067] 1.2 Fragmentation reagent: 1 tube of fragmentation buffer (120 μl / tube), 1 tube of fragmentation enzyme (120 μl / tube), 1 tube of stop reaction buffer (300 μl / tube).

[0068] 1.3 Library construction reagents: 1 tube of end repair buffer (550 μl / tube), 1 tube of end repair enzyme (27 μl / tube), 1 tube of ligation buffer (275 μl / tube), 1 tube of DNA ligase (55 μl / tube), 2 tubes of PCR enzyme mixture (1.3ml / tube), 1 tube of PCR primer mixture (137μl / tube), 1...

Embodiment 2

[0151] The detection of embodiment 2 specific samples

[0152] Select 89 enterprise reference products for preimplantation chromosomal abnormality detection, including 72 positive reference products (66 samples of different chromosome CNV sizes and trisomy cell lines and 6 samples of trisomy embryonic stem cells, a total of 72 samples), 6 A 30% chimeric reference product, 10 negative reference products, 1 data quality control reference product, 1 case of each type of sample is used as a sample to be tested, and all samples are carried out with reference to the method of using the kit in Example 1. Whole-genome amplification, fragmentation, library construction, sequencing on the machine, and simultaneous detection of negative and positive quality controls.

[0153] Test results: all negative and positive quality control products are in compliance, all positive samples in the reference product are detected, all negative samples are in compliance, all chimeric reference products...

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Abstract

The invention discloses a detection kit for chromosome abnormality before embryo implantation. The kit is characterized by comprising a single cell amplification reagent, a fragmentation reagent, a library construction reagent, a DNA purification reagent, and negative and positive quality control substances. The invention also discloses a detection method of the aforementioned kit. Compared with the prior art, the invention establishes a method using embryo blastula stage cells for preimplantation chromosome detection, adopts innovative single cell amplification technology, improves the uniformity (97% or above) and coverage (92% or above) of single cell amplification, and has the advantages of rapid detection, wide coverage, high resolution, high accuracy and moderate flux.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a preimplantation chromosome abnormality detection kit and a detection method thereof. Background technique [0002] With the pollution of our country's environment and the postponement of childbearing age, the rate of infertility is getting higher and higher. According to statistics, one out of every eight couples of childbearing age in China is unable to conceive normally, so more and more people choose assisted reproductive technology. Currently, in vitro fertilization technology represented by test-tube baby has become the mainstream of infertility treatment choose. The US Centers for Disease Control and Prevention (CDC) counted the number of cycles of assisted reproductive assisted pregnancy from 2004 to 2013. By 2013, the number of assisted reproductive cycles in the United States was close to 160,000, of which 99% were IVF cycles . But so far, even in the...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q2535/122C12Q2537/16C12Q2545/113
Inventor 梁波孔令印刘慧敏冒燕宣黎明申静静
Owner SUZHOU BASECARE MEDICAL DEVICE CO LTD
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