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

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...

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 embryos vary greatly, especially when the RNA transcript of the gene under test is low in abundance and its transcript is short-lived, while This characteristic may play a key role in the regulation of preimplantation embryonic function, and this transiently expressed gene is not easily monitored by Microarray analysis technology

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