CN112921092A - New pathogenic mutation site of fumarase gene leading to HLRCC - Google Patents

New pathogenic mutation site of fumarase gene leading to HLRCC Download PDF

Info

Publication number
CN112921092A
CN112921092A CN202110292408.0A CN202110292408A CN112921092A CN 112921092 A CN112921092 A CN 112921092A CN 202110292408 A CN202110292408 A CN 202110292408A CN 112921092 A CN112921092 A CN 112921092A
Authority
CN
China
Prior art keywords
hlrcc
cell carcinoma
renal cell
hereditary
application
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
CN202110292408.0A
Other languages
Chinese (zh)
Inventor
郑亮
张进
徐云泽
伍小宇
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.)
Shanghai Childrens Medical Center Affiliated to Shanghai Jiaotong University School of Medicine
Original Assignee
Shanghai Childrens Medical Center Affiliated to Shanghai Jiaotong University School of Medicine
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 Shanghai Childrens Medical Center Affiliated to Shanghai Jiaotong University School of Medicine filed Critical Shanghai Childrens Medical Center Affiliated to Shanghai Jiaotong University School of Medicine
Priority to CN202110292408.0A priority Critical patent/CN112921092A/en
Publication of CN112921092A publication Critical patent/CN112921092A/en
Pending legal-status Critical Current

Links

Images

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
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57438—Specifically defined cancers of liver, pancreas or kidney
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • 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
    • C12Q2600/118—Prognosis of disease development
    • 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
    • C12Q2600/136—Screening for pharmacological compounds
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/988—Lyases (4.), e.g. aldolases, heparinase, enolases, fumarase

Abstract

The invention relates to a novel pathogenic mutation site of fumarase gene causing hereditary smooth muscle sarcoidosis and renal cell carcinoma (HLRCC), comprising p.N154H, p.V156I, p.R160S, p.K414E, p.T474P, p.L207R, p.N415I, p.G280V and p.G364R. The invention provides a new molecular biology basis for the diagnosis and risk assessment of HLRCC, can develop a diagnosis kit of HLRCC based on the mutation sites to improve the accuracy of clinical diagnosis, and can also be used for risk assessment of HLRCC so as to intervene as early as possible.

Description

New pathogenic mutation site of fumarase gene leading to HLRCC
Technical Field
The invention relates to the technical field of biomedicine, in particular to a new pathogenic mutation site of fumarase gene causing HLRCC.
Background
Hereditary Leiomyomatosis and Renal Cell Carcinoma (HLRCC) syndrome, also known as Reed syndrome, is a rare renal cell carcinoma disease that was newly named and classified by the WHO world health organization in 2016. Clinically, individuals with the disease are mainly manifested as multiple skin sarcoid nodules, accompanied by type II papillary renal carcinoma and multiple uterine fibroids unique to women. In a number of reported cohort studies of HLRCC, the disease has extensive multi-organ metastasis, with metastasis occurring even in small tumors. At present, HLRCC has no effective radical cure means. The main treatment methods include surgical treatment and drug treatment. Timely surgical resection is the best method of treatment for early HLRCC, but the prognosis is often poor by the time of progression. For the advanced metastatic HLRCC, the resection cannot be performed, the traditional TKI (pezopanil, sunitinib) and mTOR (everolimus) targeted drugs for treating the advanced renal carcinoma have no obvious effect, and at present, the only relatively authoritative clinical experiment is the AVATAR research (NCT 01130519), the clinical research is that bevacizumab (bevacizumab) and erlotinib (erlotinib) are combined for treating the advanced or metastatic HLRCC, and the median progression-free survival period is 24.2 months. Immunotherapy has no significant effect on HLRCC. HLRCC has a lower five-year survival rate and a poorer prognosis than clear cell carcinoma, collecting duct carcinoma.
HLRCC is mainly caused by mutation, deletion and methylation inactivation (Loss of Function) of the fumarase gene (FH), of which about 80% of patients are germline mutations and 20% are systemic mutations. HLRCC is therefore also known as FH-deficient renal cell carcinoma. The discovery of the pathogenic mutation of HLRCC has important significance for searching a proper targeted drug to obtain better curative effect, and monitoring and prognosis of curative effect, and also can be helpful for evaluating the risk of suffering from tumor of an individual from the source so as to implement effective monitoring, early warning and intervention before suffering from the tumor. Some HLRCC pathogenic mutation sites are found at present, but the pathogenic mutation sites of the invention are not reported yet.
Disclosure of Invention
The object of the present invention is to provide new pathogenic mutation sites of fumarase gene leading to HLRCC, which addresses the deficiencies in the prior art.
In a first aspect, the invention provides an application of a reagent for specifically detecting whether one or more mutations in p.N154H, p.V156I, p.R160S, p.K414E, p.T474P, p.L207R, p.N415I, p.G280V and p.G364R exist in FH genes or proteins of a subject in preparing a kit for diagnosing hereditary smooth sarcomatosis and renal cell carcinoma or evaluating the risk of hereditary smooth sarcomatosis and renal cell carcinoma.
As a preferred example, the reagent is selected from one or more of a primer, a primer pair, a probe, an antibody and a nucleic acid chip.
In a second aspect, the present invention provides a kit comprising the reagents as described above.
In a third aspect, the invention provides the use of the kit, selected from:
1) the application in preparing products for diagnosing or assisting in diagnosing hereditary smooth muscle tumor diseases and renal cell carcinoma;
2) the application in preparing products for screening or assisting in screening hereditary smooth muscle tumor diseases and renal cell carcinoma;
3) the application in preparing products for predicting the genetic smooth muscle tumor diseases and the renal cell carcinoma risks;
4) the application in preparing products for detecting whether the gene related to hereditary smooth muscle tumor disease and renal cell carcinoma is mutated or not;
5) the application of the protein in preparing products for detecting whether the protein related to hereditary smooth sarcomatosis and renal cell carcinoma is mutated or not.
Herein, the sequence of the FH genes and proteins is as defined in GenBank ID: 2271.
The invention has the advantages that: the invention provides a new pathogenic site of HLRCC, provides a new molecular biology basis for the diagnosis and risk assessment of HLRCC, can develop a diagnosis kit of HLRCC based on the mutant sites to improve the accuracy of clinical diagnosis, and can also be used for risk assessment of HLRCC so as to intervene as early as possible.
Drawings
FIG. 1: 2-SC, FH immunohistochemical staining of G280V.
FIG. 2: 2-SC and FH immunohistochemical staining of p.G364R.
FIG. 3: immunohistochemical staining of 2-SC and FH for p.K414E.
FIG. 4: 2-SC, FH immunohistochemical staining of L207R.
FIG. 5: 2-SC, FH immunohistochemical staining of N154H.
FIG. 6: 2-SC, FH immunohistochemical staining of N415I.
FIG. 7: 2-SC, FH immunohistochemical staining of R160S.
FIG. 8: 2-SC, FH immunohistochemical staining of T474P.
FIG. 9: 2-SC, FH immunohistochemical staining of p.V156I.
FIG. 10: p.G280V, p.G364R, p.K414E, p.N154H, p.N415I, p.R160S, p.T474P, p.V156I FH-deficient renal cell carcinoma family map.
Detailed Description
The following detailed description of the present invention will be made with reference to the accompanying drawings.
Example 1
Study object and method
1. Study object
765 postoperative tissue and blood specimens of renal cancer patients are from renal cancer patients treated by renal cancer resection in urology surgery of affiliated renji hospital of Shanghai university of medical college at 1 month to 2019 month of 2008, and complete postoperative pathology confirmation report is obtained. Fixing the tissue sample by 10% formaldehyde, embedding the tissue sample in paraffin, standing the blood sample, centrifuging, subpackaging, collecting and storing in a refrigerator at minus 80 ℃. Informed consent was obtained from the patients prior to all specimen collections, and the protocol was approved and supervised by the ethical committee of the affiliated renji hospital of the Shanghai university of medicine.
2. Research method
2.1 morphological staining of H & E cells
H & E staining following conventional procedures, briefly: and sequentially carrying out dewaxing hydration, hematoxylin and eosin staining, dehydration and sealing on the paraffin section, and finally carrying out image acquisition and analysis.
2.2 immunohistochemical detection
2.2.1 immunohistochemical staining
(1) Dewaxing and hydrating: the tissue paraffin sections are placed in an incubator at 60 ℃ overnight, immersed in dimethylbenzene I and II for 15min respectively, then immersed in absolute ethyl alcohol I and II for 10min respectively, and then immersed in 90%, 80% and 70% ethyl alcohol for 10min respectively, washed with double distilled water for 3 times, 3min each time, and then immersed in a PBS solution.
(2) Antigen retrieval: the tissue slices were placed in an antigen retrieval solution containing 0.01M sodium citrate buffer (pH6.0), incubated in a 98 ℃ water bath for 15 minutes, cooled at room temperature for 30min, and washed 3 times with PBS, 3min each.
(3) Endogenous peroxidase clearance: each section was incubated with 100. mu.l of 3% hydrogen peroxide solution dropwise for 15min, followed by 3 washes in PBS for 3min each.
(4) And (3) sealing: 10% normal goat serum was added dropwise and incubated at room temperature for 1 h.
(5) Primary antibody incubation: primary antibody (FH antibody: 1:2000, ab191367, Abcam; 2SC antibody: 1:2000, crb2005017e, Discovery antibodies) was added dropwise and incubated overnight at 4 ℃.
(6) And (3) secondary antibody incubation: 200. mu.l of secondary antibody (goat anti-rabbit polyclonal antibody) was added dropwise and incubated at room temperature for 30 min.
(7) DAB color development: PBS wash 3 times, each 3 min. Adding DAB color reagent dropwise, incubating at room temperature, monitoring the color development degree under a microscope, and stopping the color development by using PBS buffer solution.
(8) Counterdyeing: counterstaining with hematoxylin for 1-3min, and washing with distilled water for 10 min.
(9) Gradient alcohol dehydration: soaking in water, 75% anhydrous ethanol, 85% anhydrous ethanol, 95% anhydrous ethanol, and 100% anhydrous ethanol for 10min respectively.
(10) Xylene soaked clear, neutral gum-encapsulated pieces, PBS substituted primary antibody as negative control.
2.2.2 evaluation of immunohistochemical staining results
All staining results were independently read by two pathologists in double-blind situations, with different results being read by more senior pathologists. Interpretation principle: 2-SC staining was divided into three intensities, 0 representing negative staining, 1+ representing focal (< 50% positive cell staining) or weaker, diffuse (> 50% positive cell staining) staining, 2+ representing diffuse (> 50% positive cell staining) staining of moderate or intensity, and negative staining of adjacent normal renal parenchyma considered as negative control. Judgment standard of dyeing result: positive (FH-,2SC + +), negative (FH +,2SC 0), and suspected positive (FH +,2SC +/+; or FH-/+,2SC + +). The number of suspected FH-deficient renal cell carcinoma patients is positive + the number of suspected positive patients.
2.3 high throughput second-generation sequencing technology for detecting Gene mutations
(1) DNA extraction and quality inspection: and (3) extracting nucleic acid from the sample DNA by using a genome DNA extraction kit, performing quality inspection on the DNA by using Nanodrop 2000, and if the sample grade is an unqualified sample, not performing the next experiment.
(2) Construction of a genomic library: and finally performing quality inspection through the steps of fragmentation/end repair/A tail addition, adaptor connection, ligation product purification, PCR amplification, product purification and the like, wherein if the grade of the sample is unqualified, the next step of experiment cannot be performed.
(3) Target area capturing: the probe marked by biotin (the gene capturing chip covers 175 genes related to metabolism, including FH gene, the chip is designed by Beijing Mikeno gene science and technology Co., Ltd.) is hybridized with library DNA under a certain condition, streptavidin modified magnetic beads are covalently combined with the biotin-marked probe, thus the target gene is captured, finally, magnetic beads carrying the target gene are adsorbed by a magnetic frame, and the target gene is eluted, purified and enriched.
(4) And (3) machine sequencing: illumina used a unique "bridge" amplification reaction, and the library was loaded onto a sequencing chip Flowcell, NextSeq 500 automated cycle and imaged. The Illumina sequencing adopts reversible terminal synthesis and sequencing reaction to sequence 2 kinds of fluorescence labeled reversible nucleotides. Only one correct complementary base can be extended in each cycle reaction, the base types are confirmed according to four different fluorescent signals, the quality of the final nucleic acid sequence is ensured, and the nucleic acid sequence is completely read after a plurality of cycles.
(5) And (3) biological information analysis: and (4) performing bioinformatics analysis on the sequencing data, and screening out a pathogenic gene and a mutation site. Further comparing and analyzing the gene sequences corresponding to the sequencing results of the patients, preferentially analyzing the insertion/deletion mutation, the nonsense mutation and the missense mutation, and dividing the results into three categories including the known mutation, the new mutation of the known gene and the new gene.
(6) Through Sanger sequencing verification, FH gene pathogenic mutation is identified: the PCR method respectively aims at the screened FH gene mutation sites and adjacent DNA sequences to carry out amplification, the primer sequences are designed by software independently developed by Beijing Mikeno gene science and technology GmbH, and NCBI online comparison tools BLAST (http:// blast.ncbi.nlm.nih.gov /) are used for eliminating false positive results and determining pathogenic mutation sites.
Second, experimental results
Through the Shanghai school of medicine of Shanghai university of transportation subsidiary Shanghai Children's medical center and Shanghai university of transportation subsidiary medical hospital, we performed H & E staining and immunohistochemical analysis of 2-SC, FH in 765 renal cell carcinoma patients' tissue sections. A total of 45 suspected FH-deficient renal cell carcinoma patients were identified by independent testing by two pathologists. Subsequently, the tissue and blood cell samples of the 45 patients were further subjected to second-generation sequencing, and finally 35 patients were diagnosed. H & E staining showed type ii papillary cell morphology, immunohistochemistry showed that 2-SC modification in FH-deficient cells stained strongly positive, and FH appeared negative, i.e. low expression. After second-generation sequencing and bioinformatic analysis, 16 mutations were found and 9 novel mutations were found (FIGS. 1-10).
According to the screening process designed by the inventor, by means of a gene chip and a deep sequencing technology, the 9 novel mutation sites of the FH gene are successfully found to be hereditary leiomyoma and renal cell carcinoma pathogenic sites.
The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and additions can be made without departing from the method of the present invention, and these modifications and additions should also be regarded as the protection scope of the present invention.

Claims (4)

1. An application of a reagent for specifically detecting whether one or more mutations in p.N154H, p.V156I, p.R160S, p.K414E, p.T474P, p.L207R, p.N415I, p.G280V and p.G364R exist in FH genes or proteins of a subject in preparing a kit for diagnosing hereditary smooth sarcomatosis and renal cell carcinoma or evaluating the risk of hereditary smooth sarcomatosis and renal cell carcinoma.
2. The reagent according to claim 1, wherein the reagent is selected from one or more of a primer, a primer pair, a probe, an antibody and a nucleic acid chip.
3. A kit comprising the reagent of claim 1.
4. Use of a kit according to claim 3, selected from the group consisting of:
1) the application in preparing products for diagnosing or assisting in diagnosing hereditary smooth muscle tumor diseases and renal cell carcinoma;
2) the application in preparing products for screening or assisting in screening hereditary smooth muscle tumor diseases and renal cell carcinoma;
3) the application in preparing products for predicting the genetic smooth muscle tumor diseases and the renal cell carcinoma risks;
4) the application in preparing products for detecting whether the gene related to hereditary smooth muscle tumor disease and renal cell carcinoma is mutated or not;
5) the application of the protein in preparing products for detecting whether the protein related to hereditary smooth sarcomatosis and renal cell carcinoma is mutated or not.
CN202110292408.0A 2021-03-18 2021-03-18 New pathogenic mutation site of fumarase gene leading to HLRCC Pending CN112921092A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110292408.0A CN112921092A (en) 2021-03-18 2021-03-18 New pathogenic mutation site of fumarase gene leading to HLRCC

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110292408.0A CN112921092A (en) 2021-03-18 2021-03-18 New pathogenic mutation site of fumarase gene leading to HLRCC

Publications (1)

Publication Number Publication Date
CN112921092A true CN112921092A (en) 2021-06-08

Family

ID=76174990

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110292408.0A Pending CN112921092A (en) 2021-03-18 2021-03-18 New pathogenic mutation site of fumarase gene leading to HLRCC

Country Status (1)

Country Link
CN (1) CN112921092A (en)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101932724A (en) * 2007-10-05 2010-12-29 环太平洋生物技术有限公司 The hyperplasia label and the prognosis of gastrointestinal cancer

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101932724A (en) * 2007-10-05 2010-12-29 环太平洋生物技术有限公司 The hyperplasia label and the prognosis of gastrointestinal cancer

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BENJAMIN B;JARISH C;ZOLTAN N;向娟;余英豪;: "皮肤平滑肌瘤免疫组化2-琥珀酸半胱氨酸(2SC)和延胡索酸水化酶(FH)染色有助于HLRCC(遗传性平滑肌瘤病及肾细胞癌综合征)的诊断", 临床与实验病理学杂志, no. 04, 28 April 2017 (2017-04-28) *
任荣鑫;赵红艺;鲍世威;张劲松;刘龙腾;: "兄弟同患多发性皮肤平滑肌瘤及其延胡索酸水合酶基因突变的分析", 中国皮肤性病学杂志, no. 02, 15 February 2017 (2017-02-15) *

Similar Documents

Publication Publication Date Title
ES2564656T3 (en) Means and methods for the non-invasive diagnosis of chromosomal aneuploidy
JP2022058469A (en) Analysis of cell-free DNA in urine and other samples
TW202012636A (en) Size-tagged preferred ends and orientation-aware analysis for measuring properties of cell-free mixtures
CN109825586A (en) DNA methylation qPCR kit and application method for lung cancer detection
WO2012060760A1 (en) Molecular marker for cancer
CN109504780A (en) DNA methylation qPCR kit and application method for lung cancer detection
CN109112216A (en) The kit and method of triple qPCR detection DNA methylations
KR20170065027A (en) Early lung cancer detection by dna methylation phenotyping of sputum-derived cells
Yan et al. Cell-of-origin subtyping of diffuse large B-cell lymphoma by using a qPCR-based gene expression assay on formalin-fixed paraffin-embedded tissues
CN110880356A (en) Method and apparatus for screening, diagnosing or risk stratification for ovarian cancer
CN109182495B (en) Gene chip and kit for noninvasive prenatal detection of bilateral goblet-shaped ear deformity
CN108342478B (en) Circulating tumor cell metabolism typing marker and application thereof
CN110714066A (en) DNA library for detecting and diagnosing corneal dystrophy disease-causing gene and application thereof
CN112921092A (en) New pathogenic mutation site of fumarase gene leading to HLRCC
CN114990218A (en) Kit for predicting lung cancer brain metastasis
CN114854853A (en) Glioma molecular typing and prognosis markers
CN105779612A (en) Lynch syndrome gene detection reagent kit and application thereof
CN110736834A (en) Method, device and system for screening and diagnosing liver cancer based on high-throughput sequencing method
CN113174440B (en) Combined gene for second-generation sequencing of multiple myeloma, application thereof and sequencing method based on combined gene
CN106834476A (en) A kind of breast cancer detection kit
WO2019062429A1 (en) Molecular marker tcons_00016233, kit using same, and application thereof
US20230079748A1 (en) Preparation method, product, and application of circulating tumor dna reference samples
US20240159742A1 (en) Epigenetic quantification using dna hybridization-based single-molecule immunofluorescent imaging
US20230203593A1 (en) Method for carrying out in vitro molecular diagnosis of ovarian tumor and kit
CN110144403B (en) New mutation SNP site of breast cancer treatment gene RBM12B and application thereof

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