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VIII type beta-tubulin gene and kit for detecting primary infertility of females

A technology of tubulin and kits, applied in genetic engineering, plant gene improvement, application, etc., can solve problems such as undiscovered female infertility

Active Publication Date: 2016-02-03
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No reports related to female infertility were found

Method used

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  • VIII type beta-tubulin gene and kit for detecting primary infertility of females
  • VIII type beta-tubulin gene and kit for detecting primary infertility of females

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Sample collection and peripheral blood DNA extraction

[0029] The patients with primary infertility caused by immature eggs came from the Shanghai Jiai Genetics and Infertility Diagnosis and Treatment Center of the Red House Hospital Affiliated to Fudan University in Shanghai, China, and the Reproductive Center of the Ninth People's Hospital Affiliated to Shanghai Jiaotong University, China. The diagnostic criteria were proposed by Rudak E et al. (Rudak E., Dor J., Kimchi M., Goldman B., Levran Dan and Mashiach S, Anomalies of humanocytes from fertile women undergoing treatment by invitrofertilization. Fertil Steril. 1990 Aug;54(2):292-6). The semen examination of the male partner was normal, while the female patient’s female reproductive organs, ovarian function, and sex hormones were all normal. More than 5 eggs were retrieved each time in more than 2 stimulation cycles, and no first polar body was visible. Under the premise of informed consent, particip...

Embodiment 2

[0030] Example 2: Detection of mutations in the TUBB8 gene

[0031] The present invention uses PCR combined with sequencing to search for TUBB8 gene mutations. The principle is to design primers (primer sequences can be specified) and amplify the four exons of the TUBB8 gene to search for mutations in the TUBB8 gene. (That is, the DNA sample is mixed with the reaction working solution (DNA polymerase, dNTP, water and buffer) and then amplified according to the PCR program. The obtained product is purified and further sequenced, and sequenced on the ABI3730 sequencer. Analysis of results Carry out by HLAFusion software (Onelambda, CA, USA, HLAFusion3.0).The PCR primer of amplification No. 1-3 exon is:

[0032] 5'-AGTTCCCAGAGGATGACCTTAGCA-3' (SEQ ID NO.2)

[0033]5'-GCTATTTAAACGTTGGCGGAGG-3' (SEQ ID NO.3)

[0034] Sequencing primers are the same as PCR primers;

[0035] The PCR primers for amplifying exon 4 are:

[0036] 5'-ACCAGGAGACCATCACACAG-3' (SEQ ID NO.4)

[0037] ...

Embodiment 3

[0041] Example 3: TUBB8 gene mutation and primary infertility caused by immature eggs

[0042] Results: We collected 7 cases of primary infertility caused by TUBB8 gene mutation. The corresponding mutation site information is shown in the table below (Table 1). For the distribution of mutation sites in the structure, see figure 1 shown.

[0043] Table 1 Patient TUBB8 gene mutation information

[0044]

[0045] In summary, the present invention has the following important practical significance:

[0046] (1) The TUBB8 gene proposed by the present invention can be used as a marker gene for predicting female infertility caused by immature eggs;

[0047] (2) The TUBB8 gene provided by the present invention can be used to evaluate or prepare a screening kit for female infertility caused by immature eggs;

[0048] (3) The TUBB8 gene provided by the present invention can be used to guide clinical patients with this gene mutation, which surgical method (IVF or ICSI) should b...

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Abstract

The invention belongs to the technical field of gene detection, and particularly discloses a VIII type beta-tubulin gene and kit for detecting primary infertility of females. The human tubulin gene-VIII type tubulin gene, namely the TUBB8 gene, is the reason for female sterility because of immature ova during mutation. By detecting the mutation of the TUBB8 gene, the TUBB8 gene can be used as a market gene for judging female sterility because of immature ova. By the adoption of the TUBB8 gene, the TUBB8 gene can be used for evaluating or preparing a screening kit for female sterility because of immature ova. The provided TUBB8 gene can be used for guiding an operational method for clinically corresponding patients during test-tube baby operations. The TUBB8 gene can be used for evaluating or preparing the screening kit with IVF / ICSI effectiveness.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a type VIII?-tubulin gene and a kit for detecting female primary infertility. Type VIII?-tubulin gene mutations are responsible for primary infertility in women. By detecting whether the gene is mutated, it can be used to diagnose and judge infertility, and to guide the patient's subsequent IVF / ICSI method or outcome. Background technique [0002] Normal pregnancy and reproduction is an important part of maintaining and continuing the human population. In clinic, microdeletion detection of AZF gene has been widely used in the diagnosis of male infertility patients (PryorJLetal, Microdeletions in the Ychromosome of infertilemen.NEnglJMed.1997Feb20;336(8);534-9). In terms of research, ZP-1, Stag3, FSHR and other genes have been found to be closely related to female infertility (HuangHLetal, MutantZP1infamilialinfertility.NEnglJMed.2014370(13):1220-6; deRouxNeta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/12C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 王磊冯睿芝桑庆胥尧
Owner FUDAN UNIV
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