CN114410777A - Renal clear cell carcinoma auxiliary immunotherapy target gene LPR8 detection kit and application thereof - Google Patents
Renal clear cell carcinoma auxiliary immunotherapy target gene LPR8 detection kit and application thereof Download PDFInfo
- Publication number
- CN114410777A CN114410777A CN202111594526.3A CN202111594526A CN114410777A CN 114410777 A CN114410777 A CN 114410777A CN 202111594526 A CN202111594526 A CN 202111594526A CN 114410777 A CN114410777 A CN 114410777A
- Authority
- CN
- China
- Prior art keywords
- lpr8
- detecting
- cell carcinoma
- clear cell
- expression level
- 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.)
- Withdrawn
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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
- C12Q1/6851—Quantitative amplification
-
- 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
- C12Q2600/158—Expression 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
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Urology & Nephrology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Pathology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Hospice & Palliative Care (AREA)
- Cell Biology (AREA)
- Oncology (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Biophysics (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Epidemiology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of biology, and relates to a renal clear cell carcinoma auxiliary immunotherapy target gene LPR8 detection kit and application thereof. The present study demonstrated a significant decrease in LPR8 expression in renal clear cell carcinoma. The expression level of LPR8 is positively correlated with immune microenvironment indexes StromalScore, ImmuneScore and ESTIMMATEScore, is positively correlated with a plurality of immune cell infiltrates and is positively correlated with the expression of immune checkpoint genes, so that LPR8 can be used as a new target for immunotherapy of renal clear cell carcinoma.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to application of a reagent for detecting an expression level of LPR8 in preparation of a preparation for renal clear cell carcinoma detection and/or adjuvant immunotherapy, and a kit for detecting a target gene LPR8 for adjuvant immunotherapy of renal clear cell carcinoma and application thereof.
Background
Renal clear cell carcinoma (KIRC) is one of the most common types of renal tumors with a relatively poor prognosis. With the development of times and the progress of modern high-throughput sequencing technologies, renal clear cell carcinoma is understood more deeply at a molecular level, but at present, the occurrence, progression and transfer mechanisms are still unclear, and effective biomarkers are lacked. Over the last decades, more and more studies have shown that the interaction between tumor cells and the microenvironment has a significant impact on treatment and prognosis. Immunotherapy is a new mode with prospect for treating renal clear cell carcinoma, has the problems of low response rate, side effect and the like, and has important significance for developing a novel auxiliary immunotherapy target point for clinical tumor immunotherapy.
Disclosure of Invention
The invention aims to provide a novel renal clear cell carcinoma immunotherapy target LPR8, and provides application of a reagent for detecting the expression level of LPR8 in preparation of a preparation for renal clear cell carcinoma detection and/or adjuvant immunotherapy, and a renal clear cell carcinoma adjuvant immunotherapy target gene LPR8 detection kit and application.
The invention provides application of an agent for detecting the expression level of LPR8 in preparing a preparation for detecting renal clear cell carcinoma and/or assisting immunotherapy.
Further, the detecting of the expression level of LPR8 comprises detecting the expression level of mRNA of LPR8 and/or detecting the expression level of protein of LPR 8.
Further, the method for detecting the expression level of LPR8 comprises the following steps: detecting the expression quantity of LPR8 mRNA in renal clear cell carcinoma and para-carcinoma tissues by an RT-qPCR method; detecting the expression level of LPR8 mRNA in renal clear cell carcinoma and para-carcinoma tissues by a molecular probe technology; the expression level of LPR8 protein in renal clear cell carcinoma and paracarcinoma tissues is detected by immunohistochemistry or Western-Blot.
More specifically, the reagent for detecting the expression level of LPR8 is an oligonucleotide probe targeting LPR8 DNA sequence or mRNA sequence, a PCR primer or an antibody targeting LPR 8.
According to the present invention, preferably, the reagent for detecting the expression level of LPR8 is SEQ ID NO: 1 and SEQ ID NO: 2 in the presence of a primer for real-time fluorescent quantitative PCR.
5’-CTTGCTGTGCATGGGAGGAT-3’(SEQ ID NO：1)
5’-AGTCCGCCCAATGTGGATTT-3’(SEQ ID NO：2)
The second aspect of the invention provides a renal clear cell carcinoma auxiliary immunotherapy target gene LPR8 detection kit, which comprises: and (3) a reagent for detecting the expression level of LPR 8.
Further, the reagent for detecting the expression level of LPR8 is an oligonucleotide probe targeting LPR8 DNA sequence or mRNA sequence, a PCR primer or an antibody targeting LPR 8.
Specifically, the reagent for detecting the expression level of LPR8 is a reagent with the nucleotide sequence shown in SEQ ID NO: 1 and SEQ ID NO: 2 in the presence of a primer for real-time fluorescent quantitative PCR.
According to the present invention, the kit may further contain other conventional reagents for real-time fluorescence quantification, and preferably, the kit further comprises at least one of the following components: trizol, isopropanol, chloroform, absolute ethanol, RNase-free water, random primers, 5 XM-MLV buffer, dNTPs, RNase inhibitor, M-MLV reverse transcriptase, a primer having the sequence of SEQ ID NO: 3 and SEQ ID NO: 4, and (4) ACTB real-time fluorescence quantitative PCR specific primers of the nucleotide sequence shown in the specification.
5’-GGCACCCAGCACAATGAAGA-3’(SEQ ID NO：3)
5’-ACTCCTGCTTGCTGATCCAC-3’(SEQ ID NO：4)
In a third aspect, the invention provides the use of the kit as described above in the preparation of a formulation for renal clear cell carcinoma detection and/or adjuvant immunotherapy.
LPR8 is a protein coding gene, and the research of the invention proves that the expression of LPR8 in renal clear cell carcinoma is obviously reduced. The expression level of LPR8 is positively correlated with immune microenvironment indexes StromalScore, ImmuneScore and ESTIMMATEScore, is positively correlated with a plurality of immune cell infiltrates and is positively correlated with the expression of immune checkpoint genes, so that LPR8 can be used as a new target for immunotherapy of renal clear cell carcinoma. And based on the result, an LPR8 detection kit is designed.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Drawings
The above and other objects, features and advantages of the present invention will become more apparent by describing in more detail exemplary embodiments thereof with reference to the attached drawings.
FIG. 1 shows the expression change of the target gene LPR8 in renal clear cell carcinoma. LPR8 expression was significantly down-regulated in renal clear cell carcinoma tissue.
Fig. 2 shows the correlation of LPR8 expression with immune cell infiltration in renal clear cell carcinoma. LPR8 expression was significantly positively correlated with multiple immune cell infiltrates.
Figure 3 shows the correlation of LPR8 expression with immune checkpoint genes in renal clear cell carcinoma. LPR8 expression was significantly positively correlated with the immune checkpoint gene.
Fig. 4 shows the expression change of LPR8 in renal clear cell carcinoma.
Detailed Description
Preferred embodiments of the present invention will be described in more detail below. While the following describes preferred embodiments of the present invention, it should be understood that the present invention may be embodied in various forms and should not be limited by the embodiments set forth herein.
The examples, in which the specific conditions are not specified, were conducted under the conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Example 1
This example serves to demonstrate that expression of the target gene LPR8 is significantly down-regulated in renal clear cell carcinoma.
LPR8 expression analysis in renal clear cell carcinoma:
1. data downloading and sorting:
TCGA renal clear cell carcinoma expression profile data and clinical data were downloaded using UCSC XENA website (https:// xenoxybrowser. net/datapages /), and LPR8 expression profile data were log2 transformed and compared between groups.
2. Differential analysis of expression of LPR8 in renal clear cell carcinoma
The difference in LPR8 mRNA levels between renal clear cell carcinoma tissue and normal tissue was analyzed using the R software package and the ggplot2 package was used for visualization as shown in fig. 1.
FIG. 1 shows the expression change of the target gene LPR8 in renal clear cell carcinoma. LPR8 expression was significantly down-regulated in renal clear cell carcinoma. P < 0.0001.
Example 2
This example demonstrates that LPR8 expression in renal clear cell carcinoma is significantly and positively correlated with tumor immune microenvironment markers.
Correlation analysis of LPR8 expression in renal clear cell carcinoma with tumor immune microenvironment:
the R software package, estamate, was used to calculate the ptorson correlation coefficient of LPR8 expression to the immune infiltration score for each renal clear cell carcinoma patient tissue, StromalScore, ImmuneScore and estamate scores. As shown in table 1.
Table 1 shows the correlation of LPR8 expression in renal clear cell carcinoma with the tumor immune microenvironment. LPR8 expression was significantly positively correlated with ESTIMATESCORE, ImmuneScore, and StromalScore.
TABLE 1
Example 3
This example was used to verify that LPR8 expression was positively correlated with immune cell infiltration in renal clear cell carcinoma.
Correlation analysis of LPR8 expression with immune cell infiltration in renal clear cell carcinoma:
and calculating a Pearson correlation coefficient of LPR8 and an immune cell infiltration score by using a corr.test function of an R software package GSVA package immune infiltration algorithm ssGSEA and psych, wherein a ggplot2 package is used for visualization. As shown in fig. 2.
Fig. 2 shows the correlation of LPR8 expression with immune cell infiltration in renal clear cell carcinoma. LPR8 expression in renal clear cell carcinoma was significantly positively correlated with multiple immune cell infiltrates.
Example 4
This example was used to verify that LPR8 expression was positively correlated with immune checkpoint gene expression in renal clear cell carcinoma.
LPR8 expression in renal clear cell carcinoma was correlated with immune checkpoint genes:
and (3) respectively calculating a Pearson correlation coefficient of LPR8 expression and an immune checkpoint gene by using a corr.test function of the R software package psych, and using ggplot2 for visualization. As shown in fig. 3.
Figure 3 shows the correlation of LPR8 expression with immune checkpoint genes in renal clear cell carcinoma. LPR8 expression was significantly positively correlated with the immune checkpoint gene.
Example 5
This example illustrates the preparation of a kit for detecting the expression level of LPR 8.
1. Primer design
PCR primers were designed based on the CDS region sequence of the transcript of the human LPR8 Gene (Gene ID:101802823), and the sequences of the primers are shown in Table 2.
2. Kit Components
(1)Trizol 50.0mL；
(2) 50.0mL of isopropanol;
(3) 50.0mL of chloroform;
(4) 50.0mL of absolute ethyl alcohol;
(5) 5.0mL of RNase-free water;
(6) 1.0. mu.M Random primers (Random primers) 50.0. mu.L;
(7) 2.0mL of 5 XM-MLV buffer;
(8)10.0mM base triphosphate deoxynucleotides (dNTPs) 100.0. mu.L;
(9) 40U/. mu.L RNase inhibitor 50.0. mu.L;
(10) 200U/. mu. L M-MLV reverse transcriptase 50.0. mu.L;
(11)ABI 2×PCR Mix 2.0mL；
(12) 10.0. mu.M LPR8 real-time fluorescent quantitative PCR specific primer 30.0. mu.L, the primer sequence is shown in Table 2;
(13) 10.0. mu.M ACTB real-time fluorescent quantitative PCR specific primers 30.0. mu.L, and the primer sequences are shown in Table 2.
TABLE 2 fluorescent quantitative RT-PCR primer sequences
3. Identification of LPR8 expression changes in renal clear cell carcinoma by RT-PCR
(1) Extraction of total RNA from tissues: the experiment was performed in an ice bath. Placing 30-50 mg of tissue (fresh or tissue preserved at-70 ℃ in liquid nitrogen) into a 1.5mL centrifuge tube, adding 1mL Trizol, homogenizing fully, and standing at room temperature for 5 min; adding 200 μ L chloroform into each tube, mixing vigorously for 30sec, standing for 15min, and centrifuging at 4 deg.C and 12000rpm for 15 min; gently sucking 400 μ L of the supernatant liquid into another new centrifuge tube, adding isopropanol with the same volume, gently inverting and mixing, and centrifuging at 12000rpm at 4 deg.C for 10 min; discarding the supernatant, adding 1ml of 75% alcohol to wash the precipitate, and centrifuging at 4 ℃ and 12000rpm for 10 min; the supernatant was discarded as much as possible, air-dried at room temperature for 10min, and dissolved by adding 10. mu.L of RNase-free water to each tube (10-15 min. lysis-promoting at 65 ℃). Determination of OD260And calculating the RNA concentration.
RNA(mg/mL)＝40×OD260X dilution multiple (n)/1000
(2) Reverse transcription: each 25. mu.L reverse transcription system contained 100pmol of random primer, 2. mu.g of total RNA, 1. mu.L of M-MLV reverse transcriptase, 0.625. mu.L of RNase inhibitor, 1.25. mu.L of dNTPs (10mM), 5. mu.L of 5 XM-MLV buffer, and 25. mu.L of RNase-free water. The reaction conditions are as follows: 1h at 37 ℃ and 5min at 95 ℃.
(3) Quantitative PCR: each 20. mu.L reaction system contained 2 XPCR Mix 10. mu.L, upstream and downstream primers 0.4. mu.L each, cDNA 1. mu.L, ddH2O8.2. mu.L. The reaction conditions are as follows: 94 ℃ for 2min, 94 ℃ for 15s, 60 ℃ for 40s, 40 cycles.
(4)2-ΔΔCTCalculating the relative expression of NSMCE 2: ACTB as reference gene, target gene C determined by qPCRTC of reference gene ACTB with the same value as that of the tissue sourceTSubtracting the values to obtain Δ CTThen will beΔCTAnd control group Δ CTSubtracting to obtain Δ Δ CT(taking a paracarcinoma sample Δ CTHas an average value of Δ CTControl), the relative expression of NSMCE2 in each group was calculated using the Power function in the Excell table. Using the software GraphPad Prism 6.0 mapping, T-test analysis of the expression differences between 9 samples of renal clear cell carcinoma (KIRC) and 9 paracarcinoma (normal), P<A difference of 0.05 is statistically significant. The results are shown in FIG. 4.
Fig. 4 shows the expression change of LPR8 in renal clear cell carcinoma. As can be seen, LPR8 expression was significantly reduced in renal clear cell carcinoma.
Having described embodiments of the present invention, the foregoing description is intended to be exemplary, not exhaustive, and not limited to the embodiments disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments.
Sequence listing
<110> Jiangsu medical profession college
<120> detection kit for renal clear cell carcinoma auxiliary immunotherapy target gene LPR8 and application
<130> BJI2103237JSYY
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
cttgctgtgc atgggaggat 20
<210> 2
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
agtccgccca atgtggattt 20
<210> 3
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
ggcacccagc acaatgaaga 20
<210> 4
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 4
actcctgctt gctgatccac 20
Claims (10)
1. The application of the reagent for detecting the expression level of LPR8 in preparing a preparation for detecting renal clear cell carcinoma and/or assisting immunotherapy.
2. The use of claim 1, wherein the detecting of the expression level of LPR8 comprises detecting the expression level of mRNA of LPR8 and/or detecting the expression level of protein of LPR 8.
3. The use of claim 1, wherein the method for detecting the expression level of LPR8 comprises: detecting the expression quantity of LPR8 mRNA in renal clear cell carcinoma and para-carcinoma tissues by an RT-qPCR method; detecting the expression level of LPR8 mRNA in renal clear cell carcinoma and para-carcinoma tissues by a molecular probe technology; the expression level of LPR8 protein in renal clear cell carcinoma and paracarcinoma tissues is detected by immunohistochemistry or Western-Blot.
4. The use of claim 1, wherein the reagent for detecting the expression level of LPR8 is an oligonucleotide probe targeting LPR8 DNA sequence or mRNA sequence, a PCR primer, or an antibody targeting LPR 8.
5. The use of claim 4, wherein the reagent for detecting the expression level of LPR8 is SEQ ID NO: 1 and SEQ ID NO: 2 in the presence of a primer for real-time fluorescent quantitative PCR.
6. A kit for detecting a renal clear cell carcinoma auxiliary immunotherapy target gene LPR8, which comprises: and (3) a reagent for detecting the expression level of LPR 8.
7. The kit of claim 6, wherein the reagent for detecting the expression level of LPR8 is an oligonucleotide probe targeting LPR8 DNA sequence or mRNA sequence, a PCR primer, or an antibody targeting LPR 8.
8. The kit according to claim 7, wherein the reagent for detecting the expression level of LPR8 is a reagent having the sequence shown in SEQ ID NO: 1 and SEQ ID NO: 2 in the presence of a primer for real-time fluorescent quantitative PCR.
9. The kit of claim 6, wherein the kit further comprises at least one of the following components: trizol, isopropanol, chloroform, absolute ethanol, RNase-free water, random primers, 5 XM-MLV buffer, dNTPs, RNase inhibitor, M-MLV reverse transcriptase, a primer having the sequence of SEQ ID NO: 3 and SEQ ID NO: 4, and (4) ACTB real-time fluorescence quantitative PCR specific primers of the nucleotide sequence shown in the specification.
10. Use of a kit according to any one of claims 6 to 9 for the manufacture of a formulation for the detection and/or adjuvant immunotherapy of renal clear cell carcinoma.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111594526.3A CN114410777A (en) | 2021-12-23 | 2021-12-23 | Renal clear cell carcinoma auxiliary immunotherapy target gene LPR8 detection kit and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111594526.3A CN114410777A (en) | 2021-12-23 | 2021-12-23 | Renal clear cell carcinoma auxiliary immunotherapy target gene LPR8 detection kit and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114410777A true CN114410777A (en) | 2022-04-29 |
Family
ID=81267259
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111594526.3A Withdrawn CN114410777A (en) | 2021-12-23 | 2021-12-23 | Renal clear cell carcinoma auxiliary immunotherapy target gene LPR8 detection kit and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114410777A (en) |
-
2021
- 2021-12-23 CN CN202111594526.3A patent/CN114410777A/en not_active Withdrawn
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109797219B (en) | Application of reagent for detecting ABRACL expression level and kit | |
| JP2010504102A5 (en) | ||
| WO2009143181A2 (en) | Micro-rna profile in human saliva and its use for detection of oral cancer | |
| CN108949992B (en) | Biomarker related to esophageal squamous carcinoma and grading thereof | |
| WO2012160551A2 (en) | Methods and compositions for determining heart failure or a risk of heart failure | |
| CN111041095B (en) | Application of reagent for detecting expression level of chromosome 8 open reading frame 73 and kit | |
| CN114410777A (en) | Renal clear cell carcinoma auxiliary immunotherapy target gene LPR8 detection kit and application thereof | |
| CN111041098B (en) | Application of reagent for detecting proline-rich and frizzled 2A expression level and kit | |
| KR101381894B1 (en) | Serum miRNA as a marker for the diagnosis of lymph node metastasis of gastric cancer | |
| CN114350810A (en) | Renal clear cell carcinoma auxiliary immunotherapy target gene TRAV18 detection kit and application thereof | |
| CN111041093B (en) | Application of reagent for detecting expression level of coiled coil domain protein 127 and kit | |
| CN114196756A (en) | Lung squamous carcinoma adjuvant immunotherapy target gene LINC00890 detection kit and application | |
| CN114350806A (en) | Lung squamous carcinoma adjuvant immunotherapy target gene C1orf162 detection kit and application thereof | |
| CN114457155A (en) | Breast cancer adjuvant immunotherapy target gene MARCHF1 detection kit and application thereof | |
| CN114350808A (en) | Breast cancer adjuvant immunotherapy target gene SHISAL2A detection kit and application thereof | |
| CN110273000B (en) | Application of reagent for detecting expression level of zinc finger protein 468 and kit | |
| CN114480641A (en) | Colorectal cancer adjuvant immunotherapy target gene NIBAN3 detection kit and application | |
| CN114574578A (en) | Lung squamous carcinoma adjuvant immunotherapy target gene C22orf15 detection kit and application thereof | |
| CN114381524A (en) | Detection kit for renal chromophobe carcinoma auxiliary immunotherapy target gene C19orf38 and application | |
| CN114350797A (en) | Colorectal cancer adjuvant immunotherapy target gene C16orf54 detection kit and application | |
| CN114277153A (en) | Application of reagent for detecting expression level of ZFP92 and kit | |
| CN114262732A (en) | Application of reagent for detecting expression level of C11orf16 and kit | |
| CN114350807A (en) | Application of reagent for detecting ZNF485 expression level and kit | |
| CN111041097A (en) | Application of reagent for detecting expression level of open reading frame 76 of chromosome 8 and kit | |
| CN114438203A (en) | Application of reagent for detecting expression level of C3orf85 and kit |
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 | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20220429 |