Construction method of gene library for detecting female infertility and kit

A technology of female infertility and gene library, which is applied in chemical library, biochemical equipment and method, microbe measurement/testing, etc., can solve the problems of high cost, long cycle, heavy workload, etc., and achieve cost reduction, Low cost and time saving effect

Pending Publication Date: 2022-08-09
REPRODUCTIVE & GENETIC HOSPITAL OF CITIC XIANGYA CO LTD
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  • Claims
  • Application Information

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Problems solved by technology

Strong pertinence also means heavy workload, long period, high cost and inevitable missed diagnosis. Therefore, the current application of first-generation PCR-Sanger sequencing to detect female infertility genetic mutations is facing great challenges

Method used

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  • Construction method of gene library for detecting female infertility and kit
  • Construction method of gene library for detecting female infertility and kit
  • Construction method of gene library for detecting female infertility and kit

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specific Embodiment

[0045] The embodiments of the present invention will be described in detail below with reference to the examples. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. In the following examples, the experimental methods that do not indicate specific conditions, preferably refer to the guidelines given in the present invention, and can also follow the experimental manuals or conventional conditions in the field, and can also follow the conditions suggested by the manufacturer, or refer to the existing conditions in the art. known experimental methods. Among them, the reagents and kits used: Peripheral blood / tissue DNA extraction kit (DNA Qiamp DNA Blood Mini Kit; 51106) from QIAGEN Company of Germany; PCR amplification kit from American Promega Company ( Green PCR Master mix: REF M7123); agar powder (TSJ001), 50×TAE electrophoresis buffer (TSG001), DNA fluorescent dye (TSJ003), and DL...

Embodiment

[0085] Example: At present, 300 clinical cases of idiopathic female infertility patients have been subjected to genetic testing and genetic etiology diagnosis (including premature ovarian failure and ovarian resistance syndrome, etc.). These patients have been excluded from exposure to toxicants, immune system diseases, infectious diseases, reproductive tract malformations, and chromosomal abnormalities. Obtain genomic DNA (peripheral blood) extraction from these patients, and take quality-controlled samples for whole-exome sequencing detection. By applying the analysis process we invented, the candidate genes selected in this project were filtered, pathogenicity analysis and family analysis were carried out, and multiple gene mutations causing primary female infertility were found in multiple unrelated families, and a series of original results. Use the following example as a template to showcase some of our results:

[0086] Homozygous mutation in SYCP2L gene was found to ...

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Abstract

The invention relates to a construction method of a next-generation sequencing gene library for detecting female infertility and a kit. The construction method comprises the following steps: obtaining genome DNA of a sample; amplifying female infertility genes in the genome DNA, and constructing a library for detecting the female infertility genes; the number of the female infertility genes is 33. One or more genes of male infertility can be amplified and a detection library can be constructed, the library can be used for next-generation sequencing, the application of the next-generation sequencing in male infertility gene detection is realized, the next-generation sequencing is adopted to detect the male infertility genes, and the kit has the advantages that (1) the flux is high; (2) the cost is low; (3) the efficiency is high; and (4) the sequencing depth is high. And (5) repeatability and traceability are realized.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a method and kit for constructing a gene library for detecting female infertility. Background technique [0002] Infertility refers to the inability of husband and wife to conceive after one year or more of unprotected cohabitation. In women (oogenesis disorders) are primarily manifested as premature ovarian insufficiency (POI) and ovarian hypoplasia, and the clinical features of the disorder include menopause before age 40 in women with decreased estrogen levels and increased gonadotropin levels, It occurs in at least 1% of women before the age of 40, and occurs in 10-28% and 4-18% of women with primary and secondary amenorrhea, respectively. [0003] At present, donor eggs and estrogen replacement therapy are the main treatment methods for female infertility patients caused by oogenesis disorders. However, in addition to ethical issues, donor eggs are also faced with the problem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12Q1/6869C12Q1/6883C40B50/06C12N15/11
CPCC12Q1/6806C12Q1/6869C12Q1/6883C40B50/06C12Q2600/16C12Q2600/156C12Q2521/101C12Q2521/501C12Q2521/514C12Q2531/113C12Q2537/143C12Q2535/122
Inventor 谭跃球蒙岚岚涂超峰何文斌袁诗敏徐西林谭琛
Owner REPRODUCTIVE & GENETIC HOSPITAL OF CITIC XIANGYA CO LTD
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