Reverse transcription reaction system for analyzing genetic expression of IVF (in-vitro fertilization) single implanting pre-embryo by reverse transcription PCR (polymerase chain reaction)

A pre-implantation embryo and gene expression technology, applied in the field of reverse transcription reaction system, can solve the problems of large differences in expression forms, inability to detect gene expression of IVF embryos as human beings, and difficulty in monitoring, etc.

Inactive Publication Date: 2013-01-09
江苏迈健生物科技发展股份有限公司
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is impossible to use Microarray analysis of 100 blastocyst-stage embryos from a single human IVF case. In other words, Microarray cannot currently be used as a means for human detection of gene expression in IVF embryos
Another important situation is that the expression patterns of different genes in oocytes and preimplantation

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  • Reverse transcription reaction system for analyzing genetic expression of IVF (in-vitro fertilization) single implanting pre-embryo by reverse transcription PCR (polymerase chain reaction)
  • Reverse transcription reaction system for analyzing genetic expression of IVF (in-vitro fertilization) single implanting pre-embryo by reverse transcription PCR (polymerase chain reaction)
  • Reverse transcription reaction system for analyzing genetic expression of IVF (in-vitro fertilization) single implanting pre-embryo by reverse transcription PCR (polymerase chain reaction)

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Embodiment Construction

[0022] Such as Figure 4 As shown, according to the purpose of the present invention, the present invention utilizes an improved real-time quantitative reverse transcription PCR analysis gene expression method in a single cell and a single pre-implantation embryo to monitor the quality of IVF embryo culture fluid, The step of using reverse transcription PCR to analyze gene expression in a single cell and a single pre-implantation embryo includes:

[0023] S1. Collect specimens: routinely collect specimens in clinical practice, wash with pre-cooled PBS solution (phosphate buffered saline) at least three times, and transfer a single fertilized egg or preimplantation embryo or embryonic cells through an ultra-fine glass pipette, with a total volume of 0.1 μL , transferred to 0.9 μL of RNA extraction solution, the extraction solution was 1×PCR buffer solution with 0.1 IU RNase inhibitor added, and all RNA was collected after liquid nitrogen treatment.

[0024] S2. The collected R...

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Abstract

The invention provides a reverse transcription reaction system for analyzing the genetic expression of an IVF (in-vitro fertilization) single implanting pre-embryo by reverse transcription PCR (polymerase chain reaction). An analysis gene expression method comprises the following steps of: collecting and purifying a specimen; carrying out reverse transcription reaction; carrying out negative control and positive control of the reverse transcription; carrying out real-time quantitative PCR detection on a reverse transcription product; optimizing a PCR condition by tissue cell RNA (ribose nucleic acid); under the condition, simultaneously amplifying and augmenting the negative control and the positive control; detecting the expression amount of a target gene with a relatively-quantitative method; and carrying out statics analysis. The improved real-time quantitative reverse transcription PCR technology is very sensitive and is highly precise, and meanwhile, considerable data can be generated. With the method, the deficiency of Microarray is made up, and the method has very wide application value for analyzing a single key gene on the expression of a single cell as well as the scientific research and the clinics of IVF. With the method, a very important monitoring means is provided for determining a new condition of IVF and culture fluid for in-vitro embryo culture.

Description

technical field [0001] The present invention relates to a gene expression analysis method of a single pre-implantation embryo of IVF (In Vitro Fertilization, test tube fertilization), in particular to an improved real-time quantitative reverse transcription PCR (Polymerase Chain Reaction, polymerase chain reaction) technology Analysis of the reverse transcription reaction system in the gene expression method of IVF single preimplantation embryos. Background technique [0002] On July 25, 1978, Louise Joy Brown, the world's first successful "test-tube" baby (IVF, test-tube fertilization), was born in England. Since then, IVF has brought good news to more than 10% of infertile couples in the world. Professor Robert Edwards, the British scientist who invented IVF, was awarded the 2010 Nobel Prize in Physiology in Medicine. However, more than three decades of clinical practice has not significantly improved the success rate of IVF. For example, in 2011, according to the stati...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 金星亮白丽君常菁
Owner 江苏迈健生物科技发展股份有限公司
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