CN117004702A - LHX8 gene for detecting female primary infertility and kit for detecting mutation of gene - Google Patents

LHX8 gene for detecting female primary infertility and kit for detecting mutation of gene Download PDF

Info

Publication number
CN117004702A
CN117004702A CN202210453448.3A CN202210453448A CN117004702A CN 117004702 A CN117004702 A CN 117004702A CN 202210453448 A CN202210453448 A CN 202210453448A CN 117004702 A CN117004702 A CN 117004702A
Authority
CN
China
Prior art keywords
seq
gene
lhx8
mutation
detecting
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202210453448.3A
Other languages
Chinese (zh)
Inventor
王磊
桑庆
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Fudan University
Zhuhai Fudan Innovation Research Institute
Original Assignee
Fudan University
Zhuhai Fudan Innovation Research Institute
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fudan University, Zhuhai Fudan Innovation Research Institute filed Critical Fudan University
Priority to CN202210453448.3A priority Critical patent/CN117004702A/en
Publication of CN117004702A publication Critical patent/CN117004702A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6858—Allele-specific amplification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medicinal Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention belongs to the technical field of gene detection, and particularly relates to an LHX8 gene for detecting female primary infertility and a kit for detecting gene mutation. The mutation of the human LHX8 gene can lead to the cause of female infertility caused by ovum maturation disorder. The mutation of the LHX8 gene can be used as a marker gene for judging female infertility caused by ovum maturation disorder. The LHX8 gene mutation provided by the invention can be used for evaluating or preparing a screening kit for female infertility caused by ovum maturation disorder. The LHX8 gene provided by the invention is mutated or not, and can be used for guiding whether a corresponding clinical patient is suitable for performing a tube infant operation.

Description

LHX8 gene for detecting female primary infertility and kit for detecting mutation of gene
Technical Field
The invention belongs to the technical field of gene detection, and particularly relates to an LHX8 gene for detecting female primary infertility and a kit for detecting gene mutation.
Background
Normal pregnancy and reproduction are important links in maintaining and continuing the human population. For female infertility, ZP1, stag3, FSHR and other genes have been found to be closely related to female infertility (Huang HL et al, mutant ZP1 in family of characteristics N Engl J Med.2014 370 (13): 1220-6;de Roux N et al, A family with hypogonadotropic hypogonadism and mutations in the gonadotropin-releasing hormone receptor. N Engl J Med. 1997 337 (22): 1597-602;Caburet S et al, mutant cohesion in premature ovarian failure. N Engl J Med.2014 370 (10): 943-9). However, none have found clinical use.
There are many causes of female infertility, and there are several clinical reports concerning descriptions of female infertility characterized by ovum immaturity (Rudak et al, fertility and sterility, 1990:292-296;Human Reproduction,1995 10:2343-2349; fertility and sterility 1999 71:567-570;Human Reproduction 2001 16 (10): 2136-2138;Human Reproduction 2002 17 (6): 1604-1609;Human Reproduction 2002 17 (10): 2556-2559). These patients failed to complete a successful in vitro insemination procedure by repeated insemination-embryo transfer (test tube infants) due to failure to obtain mature egg cells. If the related pathogenic genes can be found, the method has important significance for clinical diagnosis and typing of diseases. Recently, we have found that the pathogenic genes TUBB8 (N Engl J Med. 2016 Jan 21;374 (3): 223-32), PATL2 (Am J Hum Genet. 2017 Oct 5;101 (4): 609-615), the pathogenic genes WEE2 (Am J Hum Genet. 2018 Apr 5;102 (4): 649-657), the pathogenic genes PANX1 (Sci Transl Med. 2019 Mar 27;11 (485): eaav 8731) which lead to the egg death, the pathogenic genes BTG4 (Am J Hum Genet. 2020 Jul 2;107 (1): 24-33) and the early embryo arrest pathogenic genes PADI6 (Am J Hum Genet. 2016-752), XO43 (Hum red. 2021 Jul 19;36 (8): 92-2022), the pathogenic genes BK 4 (Am J Hum Genet. 2020 Jul 2;107 (1): 24-33), and the pathogenic genes of human egg death were responsible for a few reasons, but the reasons were still unknown in part of the patient.
Through the search of the prior art literature, only a few studies have been found around the function of the gene LHX 8. These studies report that the transcription factor of the gene specifically expressed in the early development process of the mouse ovum plays an important role in the early development of the ovum, and the homozygous knockout female mouse is completely sterile. However, no report has been made so far on the relationship between LHX8 gene mutation and human diseases, nor on the relationship with female infertility.
Disclosure of Invention
The invention aims to provide the LHX8 gene for detecting female primary infertility and the kit for detecting the mutation of the gene, which are convenient to operate and have definite effects.
The nucleic acid sequence of the LHX8 gene for detecting female primary infertility is shown as SEQ ID NO. 1.
The invention relates to a method for screening primary infertility caused by ovum maturation disorder, which is to judge whether a patient is primary infertility caused by ovum maturation disorder by detecting whether LHX8 gene is mutated or not. Patients carry LHX8 gene mutation to show infertility, ovum maturation disorder and failure of test-tube infants.
Therefore, whether the LHX8 gene is mutated or not can be used as a marker for judging female infertility caused by ovum maturation disorder.
The method for detecting whether the LHX8 gene is mutated or not can specifically detect whether the LHX8 gene is mutated or not by combining a PCR (polymerase chain reaction) method with DNA sequencing after extracting DNA from peripheral blood of a patient.
Wherein the sample to be detected is DNA, and the DNA sample is derived from peripheral blood of the population to be detected.
Alternatively, the sample to be tested is an RNA, protein, cell or serum sample from the peripheral blood of the population to be tested.
The invention relates to a primer for detecting whether LHX8 gene is mutated or not.
The LHX8 gene has 9 exon coding regions, and the PCR amplification primer pair for detecting the 1 st exon coding region is as follows:
5’- GACCTCCACTGGCGTGAATA-3’（SEQ ID NO.2）
5’- ATTCTGGTTGGCGACTCCTT-3’（SEQ ID NO.3）
sequencing primer pairs are the same;
the PCR amplification primer pair for detecting the coding regions of the No.2 and No.3 exons is as follows:
5’- AGGGCCTTCATTAGCTCGC-3’（SEQ ID NO.4）
5’- TGAGTTGCCCTAAACTGGGC-3’（SEQ ID NO.5）
sequencing primer pairs are the same;
the PCR amplification primer pair for detecting the coding region of the No.4 exon is as follows:
5’- CCACAAGGTTTCATGCTGGG-3’（SEQ ID NO.6）
5’- TCAAACAGGAAAGCACCCTCA-3’（SEQ ID NO.7）
sequencing primer pairs are the same;
the PCR amplification primer pair for detecting the coding region of the 5 th and 6 th exons is as follows:
5’- GGGGTAAGATAATGTGCTGGTT-3’（SEQ ID NO.8）
5’- CCAATAGACCCCAGGGAAACAA-3’（SEQ ID NO.9）
sequencing primer pairs are the same;
the PCR amplification primer pair for detecting the coding region of the No.7 exon is as follows:
5’- AGACTGAGCTGCTTGTTTTTAGC-3’（SEQ ID NO.10）
5’- GTAGCCTAGCCCAAGCAGGA-3’（SEQ ID NO.11）
sequencing primer pairs are the same;
the PCR amplification primer pair for detecting the coding region of the No.8 exon is as follows:
5’- GCAATTGGTCCCCTTTCCTAC-3’（SEQ ID NO.12）
5’- TTACCAGTTGCTGATTGCCT-3’（SEQ ID NO.13）
sequencing primer pairs are the same;
the PCR amplification primer pair for detecting the coding region of the No.9 exon is as follows:
5’- ATGGCAAAATCTTGTGGCCC-3’（SEQ ID NO.14）
5’- AGTGTCTTGCAGGCCAACTAT-3’（SEQ ID NO.15）
the sequencing primer pairs are as above.
The invention also relates to a method for detecting whether the LHX8 gene is mutated or not, which comprises the following steps:
for the coding region of exon 1, primer pairs were used: SEQ ID NO.2 and SEQ ID NO.3, performing PCR amplification (amplification conditions: 92 ℃ C. For 2 minutes, 92 ℃ C. For 30 seconds, 57 ℃ C. For 1 minute, 72 ℃ C. For 3 seconds (35 cycles repeated in this three step), 72 ℃ C. For 10 minutes), and then sequencing with the same primers, and comparing with the standard sequence (SEQ ID NO. 1) of LHX8 in UCSC, thereby finding a mutation;
similar to the above conditions, the coding region of exons 2 to 9 of LHX8 was amplified by the corresponding primers (SEQ ID NO.4 to 15), sequenced and aligned with the standard sequence of LHX8 in UCSC (SEQ ID NO. 1), thus finding the mutation. The LHX8 gene is tested for mutation by PCR and first generation sequencing and comparison with standard sequences.
The invention also provides a kit for detecting whether the LHX8 gene is mutated. The kit can guide doctors to judge the cause of the diseases and correctly classify the diseases, so as to inform patients of whether IVF or ICSI is adopted for the operation of the test tube infants, and whether the operation is suitable for continuous in vitro fertilization-embryo transfer (IVF) or single sperm injection (ICSI) (test tube infants).
The kit provided by the invention comprises the amplification primers SEQ ID NO.2-SEQ ID NO.15 for amplifying the 1-9 exon coding region of LHX8 and a reaction mixed solution; the reaction mixture comprises DNA polymerase, dNTP and buffer solution;
the specific using method comprises the following steps: the primers were diluted with water to a working solution at a concentration of 10 uM. The reaction system consisted of 10ul of sample DNA,0.5ul of forward primer, 0.5ul of reverse primer, 5ul of reaction mixture and 3ul of water. After mixing, a PCR reaction was performed under the above conditions, followed by a generation of sequencing.
The kit can be used for screening primary infertility caused by ovum maturation disorder.
The invention also provides a gene repair target site (namely mutation sites of actually detected patients, such as 5 mutation sites carried by 6 patients (two patients carry C412T mutation) in table 1) for researching drug treatment on primary infertility caused by ovum maturation disorder, wherein new mutation sites possibly exist in the actual detection process through comparison with a standard sequence, and LHX8 can perform normal functions through repair of the target site, so that the disease can be treated. Namely, the invention can also provide a medicament for researching the target site of repairing gene mutation, namely a medicament for treating primary infertility caused by ovum maturation disorder.
The invention can also guide patients with clinical mutation of the LHX8 gene to whether test tube babies are suitable or not by detecting the mutation of the LHX8 gene. In conventional clinics, unexplained infertility couples require test tube infants to try to reproduce offspring. However, if a patient finds by examination a mutation carrying LHX8, then the patient will be at high risk of failing the tube baby procedure. Because the patient's ovum with this mutation may not be mature, subsequent tube infant manipulations cannot be completed. Therefore, such patients are advised to donate eggs.
Drawings
FIG. 1 is a graphical representation of the distribution of patient mutations over the primary structure of LHX 8.
Detailed Description
The invention will be further illustrated by the following examples. It should be understood that the following examples are illustrative of the present invention and are not intended to limit the scope of the present invention.
Example 1: sample collection and extraction of peripheral blood DNA
Primary infertility patients come from Shanghai Jieyai and sterile diagnosis and treatment centers, shanghai Jieyai hospitals reproductive centers, shenyang essence hospitals and the like of affiliated gynaecology and obstetrics hospitals of Shanghai double denier universities. Diagnostic criteria are set forth by Rudak E et al (Rudak E., dorJ., kimchi M., goldman B., levran D and Mashiach S, anomalies of human oocytes from infertile women undergoing treatment by in vitro reference, fertillSteril 1990 Aug; 54 (2): 292-6). The semen inspection of the male is normal, the female reproductive organs, the ovary functions and the sex hormones of the female are normal, more than 5 ova are taken each time in more than 2 defecation promoting periods, most of the ova cannot be mature, or the mature ova are bad in fertilization or embryo stop is repeated. Taking part in the study of the subject on the premise of informed consent, collecting blood and signing the informed consent. All patients in the group rule out other reproductive endocrine diseases through medical history. The control population of the experiment is 1000 women with normal fertility function, 300ul of blood is sampled, DNA is extracted according to the instruction of the kit, and the concentration and purity of the DNA are detected by an ultraviolet spectrophotometer.
Example 2: detection of mutation of LHX8 Gene
The invention adopts PCR combined sequencing to search LHX8 gene mutation. The principle is that the primer design (the primer sequence can be specifically given) and the amplification are carried out on 9 exon coding regions of the LHX8 gene, and the LHX8 gene mutation is searched. (i.e., mixing the DNA sample with reaction working solution (DNA polymerase, dNTP, water and buffer) and amplifying according to PCR procedure. The obtained product is purified and further sequenced by sequencing on ABI 3730. The analysis of the result is performed by HLA Fusion software (One lambda, CA, USA, HLA Fusion 3.0).
Example 3: LHX8 gene mutation and primary infertility caused by ovum maturation disorder
Results: we have collected 6 cases of primary infertility patients (two of which carry C412T mutation) caused by ovum maturation disorder due to LHX8 gene mutation. The corresponding mutation site information is shown in the following table (Table 1). The structural distribution of the mutation sites is shown in FIG. 1.
TABLE 1 patient LHX8 Gene mutation information
。
In summary, the invention has the following important practical significance:
(1) The LHX8 gene provided by the invention can be used as a marker gene for predicting female infertility caused by ovum maturation disorder;
(2) The LHX8 gene provided by the invention can be used for evaluating or preparing a screening kit for female infertility caused by ovum maturation disorder;
(3) The invention can be used for researching and preparing medicines for repairing gene mutation target sites, namely medicines for treating primary infertility caused by ovum maturation disorder;
(4) The LHX8 gene provided by the invention can be used for guiding patients with clinical gene mutation, judging the success possibility of test tube infants and suggesting the subsequent egg donation.
Sequence listing
<110> university of double denier
Zhuhai Fudan Innovation Research Institute
<120> LHX8 gene for detecting female primary infertility and kit for detecting mutation of the gene
<130> 001
<160> 15
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1071
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 1
atgcagattc tgagcaggtg tcaggggctc atgtcagagg agtgcgggcg gactacagcc 60
ctggcggccg ggaggactcg caaaggcgcc ggggaagagg gactggtgag ccccgaggga 120
gcgggggacg aggactcgtg ctcctcctcg gccccgctgt ccccgtcgtc ctcgccccgg 180
tccatggcct cgggctccgg ctgccctcct ggcaagtgtg tgtgcaacag ttgcggcctg 240
gagatcgtgg acaaatacct tctcaaggtg aatgacctat gctggcatgt ccggtgtctc 300
tcctgcagtg tttgcagaac ctccctagga aggcacacca gctgttatat taaagacaaa 360
gacattttct gcaaacttga ttatttcaga aggtatggaa ctcgctgctc tcgatgtggg 420
agacacatcc attctactga ctgggtccgg agagccaagg ggaatgtcta tcacttggca 480
tgctttgcct gcttttcctg caaaaggcaa ctttccacag gagaggagtt tgctttggtg 540
gaagagaaag tcctctgcag agtacattat gactgcatgc tggataattt aaaaagagaa 600
gtagaaaatg ggaatgggat tagtgtggaa ggtgccctcc tcacagagca agatgttaac 660
catccaaaac cagcaaaaag agctcggacc agctttacag cagatcagct tcaggttatg 720
caagcacaat ttgctcagga caacaaccca gatgcacaga cactccagaa attggcagaa 780
aggacaggct tgagcagacg tgtgatacag gtgtggtttc agaattgtag agcacgccac 840
aagaaacacg tcagtcctaa tcactcatcc tccaccccag tcacagcagt cccaccctcc 900
aggctgtctc cacccatgtt agaagaaatg gcttattctg cctacgtgcc ccaagatgga 960
acgatgttaa ctgcgctgca tagttatatg gatgctcatt caccaacaac tcttggactc 1020
cagcccttgt taccccattc aatgacacaa ctgccaataa gtcataccta a 1071
<210> 2
<211> 20
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 2
gacctccact ggcgtgaata 20
<210> 3
<211> 20
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 3
attctggttg gcgactcctt 20
<210> 4
<211> 19
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 4
agggccttca ttagctcgc 19
<210> 5
<211> 20
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 5
tgagttgccc taaactgggc 20
<210> 6
<211> 20
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 6
ccacaaggtt tcatgctggg 20
<210> 7
<211> 21
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 7
tcaaacagga aagcaccctc a 21
<210> 8
<211> 22
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 8
ggggtaagat aatgtgctgg tt 22
<210> 9
<211> 22
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 9
ccaatagacc ccagggaaac aa 22
<210> 10
<211> 23
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 10
agactgagct gcttgttttt agc 23
<210> 11
<211> 20
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 11
gtagcctagc ccaagcagga 20
<210> 12
<211> 21
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 12
gcaattggtc ccctttccta c 21
<210> 13
<211> 20
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 13
ttaccagttg ctgattgcct 20
<210> 14
<211> 20
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 14
atggcaaaat cttgtggccc 20
<210> 15
<211> 21
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 15
agtgtcttgc aggccaacta t 21

Claims (5)

1. The application of LHX8 gene as the mark for judging female primary sterility caused by ovum maturation disorder, and the nucleic acid sequence of LHX8 gene is shown in SEQ ID NO. 1.
2. A method for screening female primary infertility caused by ovum maturation disorder, which is characterized in that whether a patient is primary infertility caused by ovum maturation disorder is judged by detecting whether LHX8 gene is mutated.
3. A primer for detecting whether or not mutation is generated in the LHX8 gene, wherein the PCR amplification primer pair for detection is sequentially: SEQ ID No.2, SEQ ID No.3, SEQ ID No.4, SEQ ID No.5, SEQ ID No.6, SEQ ID No.7, SEQ ID No.8, SEQ ID No.9, SEQ ID No.10, SEQ ID No.11, SEQ ID No.12, SEQ ID No.13, SEQ ID No.14, SEQ ID No.15.
4. A method for detecting whether mutation occurs in LHX8 gene, which is characterized by comprising the following specific steps:
and (3) carrying out PCR amplification on 9 exon coding regions in LHX8 by using corresponding primer pairs SEQ ID NO.2, SEQ ID NO. 3-SEQ ID NO.14 and SEQ ID NO.15 respectively, and then sequencing by using the same primer to obtain a standard sequence of LHX8 in UCSC: the SEQ ID NO.1 was aligned so that mutations were found.
5. A kit for detecting the occurrence of a mutation in the LHX8 gene, the kit comprising amplification primers for amplifying the coding region of exons 1 to 9 of the LHX8 gene and corresponding sequencing primers: SEQ ID NO.2-SEQ ID NO.15, and a reaction mixture comprising DNA polymerase, dNTPs and a buffer.
CN202210453448.3A 2022-04-27 2022-04-27 LHX8 gene for detecting female primary infertility and kit for detecting mutation of gene Pending CN117004702A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202210453448.3A CN117004702A (en) 2022-04-27 2022-04-27 LHX8 gene for detecting female primary infertility and kit for detecting mutation of gene

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202210453448.3A CN117004702A (en) 2022-04-27 2022-04-27 LHX8 gene for detecting female primary infertility and kit for detecting mutation of gene

Publications (1)

Publication Number Publication Date
CN117004702A true CN117004702A (en) 2023-11-07

Family

ID=88562295

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202210453448.3A Pending CN117004702A (en) 2022-04-27 2022-04-27 LHX8 gene for detecting female primary infertility and kit for detecting mutation of gene

Country Status (1)

Country Link
CN (1) CN117004702A (en)

Similar Documents

Publication Publication Date Title
Pandey et al. Loss of the AZFc region due to a human Y-chromosome microdeletion in infertile male patients
JP2004528027A (en) Diagnostic test
CN107475378B (en) PATL2 gene for detecting female primary infertility and kit for detecting gene mutation
CN111549036A (en) Marker BTG4 gene for judging female primary infertility and detection kit thereof
CN109055532A (en) Hereditary hearing impairment genetic test Primer composition, kit and application before embryo implantation
CN105296634B (en) For detecting the VIII type &#39;beta &#39;-tubulin gene and kit of women primary infertility
CN117248030A (en) PKD1 variant molecule detection method based on single-cell whole genome amplification and application thereof
CN109504761A (en) It detects the PANX1 gene of women primary infertility and detects the kit of the gene mutation
CN117004702A (en) LHX8 gene for detecting female primary infertility and kit for detecting mutation of gene
CN112877414A (en) Marker CDC20 gene for judging female primary infertility and detection kit thereof
CN112877415A (en) Marker TRIP13 gene for judging female primary infertility and detection kit thereof
CN116042798A (en) TUBA4A gene for detecting female primary infertility and kit for detecting mutation of gene
CN117004703A (en) KPNA7 gene for detecting female primary infertility and kit for detecting mutation of KPNA7 gene
Song et al. Genome-wide screening of severe male factor infertile patients using BAC-array comparative genomic hybridization (CGH)
KR20080082562A (en) Diagnostic methods for male infertility and diagnostic kits for male infertility for detecting y chromosomal microdeletion
JPH11507222A (en) Detection of male infertility Y chromosome deletion by multiplex primer binding
CN115927584A (en) TACC3 gene for detecting female primary infertility and kit for detecting mutation of gene
Grifo et al. Update in preimplantation genetic diagnosis: successes, advances, and problems
CN115896269A (en) PABPC1L gene and kit for detecting primary infertility caused by ovum maturation disorder
Wang et al. De novo paternal FBN1 mutation detected in embryos before implantation
KR100908691B1 (en) 남성 Male infertility diagnostic kit for detecting chromosomal microdeletion
CN113136423A (en) FBXO43 gene for detecting female primary infertility and kit for detecting gene mutation
Fu et al. Multiple displacement amplification as the first step can increase the diagnostic efficiency of preimplantation genetic testing for monogenic disease for β‐thalassemia
CN113322311A (en) MEI1 gene for detecting female primary infertility and kit for detecting mutation of gene
CN112609012A (en) Real-time fluorescent PCR (polymerase chain reaction) primer, probe and kit for detecting group B streptococcus nucleic acid

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination