CN117089618A - Application of AZGP1 as prognosis risk marker for patients with multiple myeloma - Google Patents

Application of AZGP1 as prognosis risk marker for patients with multiple myeloma Download PDF

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CN117089618A
CN117089618A CN202311089468.8A CN202311089468A CN117089618A CN 117089618 A CN117089618 A CN 117089618A CN 202311089468 A CN202311089468 A CN 202311089468A CN 117089618 A CN117089618 A CN 117089618A
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multiple myeloma
azgp1
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patients
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黄晓军
阮国瑞
廖明玥
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Peking University Peoples Hospital
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Abstract

The application discloses application of AZGP1 as a prognosis risk marker for patients with multiple myeloma. The application analyzes the expression level of AZGP1 in bone marrow cells of a first diagnosis and alleviation MM patient and a healthy donor by an RT-qPCR method, and discovers that the expression level of the gene in the MM patient is obviously higher than that of a normal control and possibly related to clinical course. The clinical significance of the gene was then explored and high expression of the gene was found to be a risk factor for low Progression Free Survival (PFS) in MM patients. AZGP1 has important value in the diagnosis and prognosis of multiple myeloma.

Description

Application of AZGP1 as prognosis risk marker for patients with multiple myeloma
Technical Field
The application belongs to the technical field of medical detection, and particularly relates to application of AZGP1 as a marker for evaluating prognosis risks of patients with multiple myeloma.
Background
Multiple Myela (MM) is a malignancy common to the blood system and is characterized by abnormal proliferation of clonal plasma cells and secretion of large amounts of nonfunctional immunoglobulins or fragments thereof, common symptoms including hypercalcemia, impairment of renal function, anemia, bone disease, secondary amyloidosis, and the like. With the continuous advent of new drugs and the improvement of detection means, the diagnosis and treatment of MM are continuously improved and perfected, but the recurrence and death of almost all patients are still unavoidable, and currently, the patients cannot be cured. Reproducible cytogenetics and molecular abnormalities are important factors in the development of MM, and in addition, the presence of minimal residual lesions is a major cause of their recurrence. There is still a lack of molecular targets that can be easily and sensitively detected and specifically diagnosed, relapse pre-warned, and prognostic assessed.
AZGP1 (Alpha-2-glycopin 1) is localized on human chromosome 7q22.1 and its encoded zinc-Alpha 2-Glycoprotein is secreted in various body fluids and is expressed mainly in epithelial cells of mammary gland, prostate, liver and various gastrointestinal organs, and actively participates in important functions of many human bodies including fertilization, immunoregulation, lipid mobilization, and the like.
Disclosure of Invention
The application aims to provide an AZGP1 biomarker related to diagnosis and prognosis of multiple myeloma and application thereof.
In order to achieve the above object, the present application provides a novel use of a substance for detecting the expression level of AZGP 1.
The present application provides the use of a substance for detecting the expression level of AZGP1 in any one of the following (a 1) to (a 6):
(a1) Preparing a product for diagnosing or aiding in diagnosing multiple myeloma;
(a2) Preparing a product for screening or assisting in screening patients with multiple myeloma;
(a3) Preparing a product for assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
(a4) Preparing a product for assessing or aiding in assessing the progression-free survival of a patient with multiple myeloma;
(a5) Preparing a product for assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(a6) Products are prepared for or aiding in the assessment of progression of disease in patients with multiple myeloma.
The application also provides application of the substances for detecting the AZGP1 expression level in any one of the following (b 1) - (b 6):
(b1) Diagnosing or aiding in the diagnosis of multiple myeloma;
(b2) Screening or aiding in screening patients with multiple myeloma;
(b3) Assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
(b4) Assessing or aiding in assessing progression-free survival of a patient with multiple myeloma;
(b5) Assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(b6) The disease course of the patients with the multiple myeloma is evaluated or assisted in evaluation.
In order to achieve the above object, the present application also provides a kit having the function of any one of the following (c 1) to (c 6):
(c1) Diagnosing or aiding in the diagnosis of multiple myeloma;
(c2) Screening or aiding in screening patients with multiple myeloma;
(c3) Assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
(c4) Assessing or aiding in assessing progression-free survival of a patient with multiple myeloma;
(c5) Assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(c6) The disease course of the patients with the multiple myeloma is evaluated or assisted in evaluation.
The kit provided by the application comprises a substance for detecting the AZGP1 expression quantity.
In any of the above applications or kits, the substance for detecting the expression level of AZGP1 may be a substance for detecting the expression level of AZGP1 gene, a substance for detecting the expression level of mRNA encoded by AZGP1 gene, or a substance for detecting the content of protein encoded by AZGP1 gene.
Further, the substance for detecting the expression level of the AZGP1 gene may be a reagent and/or an instrument required for detecting the expression level of the AZGP1 gene by a method in the prior art, such as a reagent and/or an instrument required for detecting the expression level of the gene by high throughput sequencing, or a reagent and/or an instrument required for detecting the expression level of the gene by quantitative PCR, or a reagent and/or an instrument required for detecting the expression level of the gene by a northern hybridization technique.
The substance for detecting the protein content encoded by the AZGP1 gene may be a reagent and/or an instrument required for detecting the protein content by a method in the prior art, such as a reagent and/or an instrument required for detecting the protein content by mass spectrometry or a related technique thereof, or a reagent and/or an instrument required for detecting the protein content by an immunohybridization technique, or a reagent and/or an instrument required for detecting the protein content by an ELISA technique, or a reagent and/or an instrument required for detecting the protein content by a protein chip or test paper.
The substance for detecting the mRNA expression amount encoded by the AZGP1 gene can comprise a specific amplification primer and/or a probe for detecting the mRNA of the AZGP1 gene. In a specific embodiment of the application, the specific amplification primers used for detecting the mRNA of the AZGP1 gene consist of a single-stranded DNA molecule shown in SEQ ID No.1 and a single-stranded DNA molecule shown in SEQ ID No. 2. The probe for detecting the mRNA of the AZGP1 gene is a single-stranded DNA molecule shown as SEQ ID No.3, wherein the 5 'end of the probe can be marked with a fluorescence reporting group (such as FAM), and the 3' end of the single-stranded probe can be marked with a fluorescence quenching group (such as BHQ).
Still further, the expression level is a relative expression level of the reference gene of the AZGP1 gene, and specifically may be a ratio of the expression level of the AZGP1 gene to the expression level of the reference gene. The expression level of the AZGP1 gene and the expression level of the reference gene can be copy numbers obtained according to standard curves and CT values.
Further, the substance for detecting the expression amount of mRNA encoded by the AZGP1 gene can further comprise a specific amplification primer and/or a probe for detecting mRNA of the reference gene. The reference gene may specifically be the ABL1 gene. In one embodiment of the present application, the specific amplification primers for detecting the ABL1 gene mRNA consist of a single-stranded DNA molecule shown in SEQ ID No.4 and a single-stranded DNA molecule shown in SEQ ID No. 5; the probe for detecting ABL1 gene mRNA is a single-stranded DNA molecule shown in SEQ ID No.6, the 5 'end of the probe can be marked with a fluorescence reporting group (such as FAM), and the 3' end of the single-stranded probe can be marked with a fluorescence quenching group (such as BHQ).
The kit can also comprise a data processing device A; a module is arranged in the data processing device A; the module has the following functions: diagnosing whether the testee is a multiple myeloma patient according to the relative expression quantity of AZGP1 in the bone marrow mononuclear cells of the testee: if the relative expression amount of AZGP1 in the bone marrow mononuclear cells of the tested person is larger than that of the healthy control person, the tested person is or is suspected to be a patient with multiple myeloma; otherwise, the subject is not or suspected of not being a multiple myeloma patient.
The kit may further comprise a data processing device B; a module is arranged in the data processing device B; the module has the following functions: assessing the prognostic risk of a multiple myeloma patient based on the relative expression level of AZGP1 in bone marrow mononuclear cells of the multiple myeloma patient: the risk of prognosis for patients with multiple myeloma in AZGP 1-low expression group is lower than or the candidate for patients with multiple myeloma in AZGP 1-high expression group.
The kit may further comprise a data processing device C; the data processing device C is internally provided with a module; the module has the following functions: assessing progression-free survival of multiple myeloma patients based on the relative expression level of AZGP1 in bone marrow mononuclear cells of multiple myeloma patients: the progression free survival rate of patients with multiple myeloma in AZGP1 low expression group is higher than or candidates for patients with multiple myeloma in higher AZGP1 expression group.
The kit may further comprise a data processing device D; a module is arranged in the data processing device D; the module has the following functions: assessing the progression-free survival time of the multiple myeloma patients according to the relative expression amount of AZGP1 in bone marrow mononuclear cells of the multiple myeloma patients: the progression free survival time of patients with multiple myeloma in AZGP1 low expression group is longer than or candidates for patients with multiple myeloma longer than that in AZGP1 high expression group.
The kit may further comprise a data processing device D; a module is arranged in the data processing device D; the module has the following functions: assessing the disease course progress of the multiple myeloma patients according to the relative expression amount of AZGP1 in bone marrow mononuclear cells of the multiple myeloma patients: multiple myeloma patients in the AZGP 1-low expression group have a lower risk of relapse than or a candidate for multiple myeloma patients in the AZGP 1-high expression group.
The above AZGP1 high expression group and the AZGP1 low expression group can be determined as follows: taking isolated bone marrow of a group to be detected consisting of a plurality of patients with multiple myeloma without any treatment measures as a specimen, measuring the relative expression quantity of AZGP1 in each specimen, arranging and trisecting the group to be detected according to the sequence of the relative expression quantity from low to high, taking 1/3 of the group to be detected with low expression quantity as an AZGP1 low expression group, and taking the rest 2/3 of the groups to be detected as an AZGP1 high expression group.
The use of AZGP1 as a biomarker in any of the following (a 1) - (a 6) is also within the scope of the application:
(a1) Preparing a product for diagnosing or aiding in diagnosing multiple myeloma;
(a2) Preparing a product for screening or assisting in screening patients with multiple myeloma;
(a3) Preparing a product for assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
(a4) Preparing a product for assessing or aiding in assessing the progression-free survival of a patient with multiple myeloma;
(a5) Preparing a product for assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(a6) Products are prepared for or aiding in the assessment of progression of disease in patients with multiple myeloma.
In any one of the above applications or products, the nucleotide sequence of the AZGP1 gene is shown in SEQ ID No.9 (NCBI Reference Sequence: NM-001185.4).
In the present application, the prognostic risk is a prognostic risk of a patient with primary diagnosis of multiple myeloma. The prognosis risk can be embodied as all or part of the following indexes: progression free survival time, progression free survival rate. The longer the progression free survival, the lower the risk of prognosis; the higher the progression free survival, the lower the risk of prognosis.
The disease progression (recurrence risk) refers to the recurrence risk of patients with multiple myeloma after remission.
In any of the above applications or products, the screening or aiding in screening of multiple myeloma patients includes screening or aiding in screening of primary multiple myeloma patients and screening or aiding in screening of relapsing multiple myeloma patients.
In any of the above applications or products, the multiple myeloma is adult multiple myeloma.
According to the application, the RT-qPCR method is used for analyzing the initial diagnosis and relieving the AZGP1 expression level in bone marrow mononuclear cells of MM patients, and the high AZGP1 expression is found to be a risk factor of low Progression Free Survival (PFS) of MM patients for the first time by combining clinical course and survival data. AZGP1 has important value in the diagnosis and prognosis of multiple myeloma.
Drawings
FIG. 1 shows the level of AZGP1 expression in bone marrow cells of a first diagnosis and alleviation of MM patients and healthy donors. Lines represent median and quartile values; * P < 0.0001.
Fig. 2 is a ROC graph of AZGP1 diagnostic first diagnosis MM.
FIG. 3 shows the relationship between AZGP1 expression level and MM clinical course. Expression levels of AZGP1 in bone marrow single nuclear cell specimens of initial diagnosis, remission and disease progression in 7 MM patients.
FIG. 4 shows the relationship between AZGP1 expression level and the progression-free survival of MM prognosis.
Detailed Description
The following detailed description of the application is provided in connection with the accompanying drawings that are presented to illustrate the application and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the application in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
The nucleotide sequence of the AZGP1 gene is shown as SEQ ID No.9 (NCBI Reference Sequence: NM_ 001185.4).
Example 1 use of AZGP1 as a marker for assessing prognosis risk in patients with multiple myeloma
1. Study object and method
1. Study object
Bone marrow specimens collected from 147 cases of primary multiple myeloma patients from the institute of hematopathy in the civil hospital at Beijing university during the period of 1 month to 12 months in 2014 were taken as subjects. 147 multiple myeloma patients included 93 men, 54 women, with a median age of 59 years and an age range of 33-87 years, and were followed until death, no follow-up, or 8 months of 2019. The diagnostic criteria for multiple myeloma refer to guidelines of the national integrated cancer network (NCCN), and the staging is performed according to the Durie-Salmon (D-S) staging system, the International Staging System (ISS) and the revised International staging system (R-ISS). Treatment and efficacy assessments for patients with multiple myeloma are made with reference to the guidelines above. Progression Free Survival (PFS) is defined as the time from the onset of primary treatment to the first occurrence of disease progression, which is an event.
In addition, a total of 131 bone marrow specimens were collected during follow-up of 95 primary patients with multiple myeloma, including 28 bone marrow specimens that were treated to obtain Complete Remission (CR), 31 bone marrow specimens that were treated to obtain Very Good Partial Remission (VGPR), 36 bone marrow specimens that were treated to obtain Partial Remission (PR), and 36 bone marrow specimens that were treated to develop disease progression. Normal control bone marrow specimens were obtained from adult healthy volunteers for a total of 47 cases.
Specimens used in this study protocol were approved by the ethical committee of the civil hospital at the university of Beijing. All healthy volunteers and patients signed informed consent.
2. Bone marrow mononuclear cell extraction and RT-qPCR
Mononuclear cells in bone marrow specimens were separated using Ficoll lymphocyte separation medium and density gradient centrifugation, and RNA was extracted and reverse transcribed into cDNA. Using cDNA as template, adopting AZGP1 primer pair (AZGP 1 primer pair is formed from AZGP1 upstream primer and AZGP1 downstream primerThe size of the amplified product is 95bp, the nucleotide sequence is shown as SEQ ID No. 7) and the AZGP1 probe are subjected to RT-qPCR. And (3) taking cDNA as a template, and performing RT-qPCR by using an ABL1 primer pair (wherein the ABL1 primer pair consists of an ABL1 upstream primer and an ABL1 downstream primer, the amplification product is 124bp in size, and the nucleotide sequence is shown as SEQ ID No. 8) and an ABL1 probe. The following 10 μl PCR reaction system was configured using PCR Master Mix kit:universal PCR Master Mix; upstream primer 0.9. Mu.M, downstream primer 0.9. Mu.M, probe 0.25. Mu.M; 150-500ng cDNA, primer sequences and fluorescent probe sequences are shown in Table 1. qPCR was performed using an ABI 7500FAST PCR amplification apparatus under the following reaction conditions: 50℃2min,95℃10min, then 95℃15s,60℃1min for 40 cycles. And calculating copy numbers of AZGP1 and ABL1 by a standard curve method by taking ABL1 as an internal reference. Serial dilution (10) 6 、10 5 、10 4 、10 3 、10 2 、10 1 And 10 0 Copy/. Mu.L) of ABL1 expressing plasmids (see "Gabert J, beillillard E, van der Velden VH, bi W, grimwade D, pallysgaard N, et al Standard and quality control studies of" real-time "quantitative reverse transcriptase polymerase chain reaction of fusion gene transcripts for residual disease detection in leukemia-a Europe Against Cancer program. Leukemia.2003;17:2318-2357, "ABL1 plasma" as used herein) and AZGP1 positive bone marrow specimens were used to construct standard quantitative curves. The curve threshold is set to 0.082. Obtaining Ct values of ABL1 and AZGP1 through amplification curves of sample internal reference genes ABL1 and AZGP1 genes and a set threshold (0.082), obtaining copy numbers of sample ABL1 and AZGP1 according to an ABL1 standard curve (because the amplification efficiency of AZGP1 and ABL1 is similar, and the sample ABL1 standard curve is referred to for calculation in order to reduce experimental errors), dividing the copy number of AZGP1 by the copy number of ABL1 to obtain the expression quantity of sample AZGP1, multiplying the result by one hundred percent for the consistency with the form reported by clinical routine, and finally, obtaining the result in the form of (the copy number of AZGP 1/the copy number of ABL 1) ×percent=the expression quantity of sample AZGP 1.
Table 1, AZGP1 and ABL1 primer and probe sequences
Primer(s) Sequence (5 '-3')
AZGP1 upstream primer 5'-AGACCCTGAAAGACATCGTGGA-3’(SEQ ID No.1)
AZGP1 downstream primer 5'-GCTTCTGTTATTCTCGATCTCACAAC-3’(SEQ ID No.2)
AZGP1 probe 5'-FAM-CAACGACAGTAACGGGTCTCACGTATTGC-BHQ-3’(SEQ ID No.3)
ABL1 upstream primer 5'-TGGAGATAACACTCTAAGCATAACTAAAGGT-3’(SEQ ID No.4)
ABL1 downstream primer 5'-GATGTAGTTGCTTGGGACCCA-3’(SEQ ID No.5)
ABL1 probe 5'-FAM-CCATTTTTGGTTTGGGCTTCACACCATT-BHQ-3’(SEQ ID No.6)
3. Statistical analysis
Statistical analysis was performed using SPSS26.0, R software 4.2.1, graphpad Prism 8.0.2. And comparing the difference of the two groups of data, wherein the classification variable data adopts chi-square test, the continuous variable data adopts t test, and the difference is less than 0.05, so that the statistical significance is realized. Subject work (receiver operating characteristic, ROC) curves were used to evaluate the specificity and sensitivity of diagnostic indicators. The Jooden Index (Youden Index) is used to calculate the cut-off value for the diagnosis. Survival analysis was performed using Kaplan-Meier using log-rank test. The Cox proportional hazards regression model performs multi-factor analysis. The variable with P more than 0.1 is gradually removed by a back-off method, and P less than 0.05 has statistical significance.
2. Results of the study
1. Expression level of AZGP1 in multiple myeloma
147 primary multiple myeloma patients, 95 multiple myeloma remission patients and 47 healthy donor control bone marrow specimens were analyzed for AZGP1 expression levels.
The results show that the expression level of AZGP1 in bone marrow cells of primary patients with multiple myeloma (median of 1.91%, range of 0-3305.45%) is significantly higher than that of patients with multiple myeloma relief (median of 0.14%, range of 0-160.84%, P < 0.0001) and healthy donor controls (median of 0.08%, range of 0-0.47%, P < 0.0001). While there was no significant difference in expression levels between the multiple myeloma-alleviating patients and healthy donor controls (p=0.378) (fig. 1).
ROC curve analysis showed that the area under the curve (area under the curve, AUC) of AZGP1 diagnostic MM was 0.756 (95% confidence interval 0.690-0.822; p < 0.0001; fig. 2), with a maximum approximate log index corresponding to an AZGP1 expression level of 0.48%. With this as the diagnostic threshold for MM, the over-expression rate of AZGP1 in the initial MM was 58.50%.
2. AZGP1 expression level is related to clinical course of multiple myeloma
To further investigate the relationship between AZGP1 and the clinical course of adult multiple myeloma, the analysis of AZGP1 expression levels was performed on bone marrow specimens from 7 adult multiple myeloma patients at initial diagnosis, complete remission and disease progression.
The results showed that the level of AZGP1 expression in bone marrow cells was significantly reduced compared to the initial bone marrow cells at complete remission, whereas the level of AZGP1 expression in bone marrow cells was significantly increased compared to complete remission at relapse (disease progression) (fig. 3).
3. Relationship between AZGP1 expression level and general clinical characteristics of multiple myeloma
147 cases of patients initially diagnosed with multiple myeloma were classified into AZGP1 high-expression groups and AZGP1 low-expression groups according to the following methods according to the AZGP1 expression levels: determining the relative expression quantity of AZGP1 in each patient bone marrow specimen, then arranging and trisecting the groups to be tested according to the sequence from low to high of the relative expression quantity of AZGP1, taking 1/3 of the groups to be tested with low relative expression quantity as AZGP1 low expression groups, taking the rest 2/3 of the groups to be tested as AZGP1 high expression groups, and analyzing the relation between the expression level of AZGP1 and the general clinical characteristics of multiple myeloma.
The results show that at initial diagnosis, AZGP1 high-expression group patients are more prone to develop stages IIIA and IIIB (94.8% vs.72.9%, p=0.001), bone destruction at 3 and above (65.2% vs.45.8%, p=0.042), and bone marrow plasma cell fraction is higher (p=0.011), serum light chain and monoclonal antibody levels are higher (p=0.012, p=0.009), and peripheral platelet count is lower (p=0.018), and low serum albumin levels (< 35g/L,65.6% vs.40.0%, p=0.005) are more likely to occur compared to AZGP1 low-expression group patients. In addition, patients in AZGP 1-high expressing groups are more prone to high risk cytogenetic markers associated with poor prognosis (42.0% vs.5.6%, P < 0.001). The remaining indices of both groups were not statistically different (table 2).
Table 2, relationship between AZGP1 expression level and general clinical characteristics of adult multiple myeloma in initial diagnosis
4. Relationship between AZGP1 expression level and prognosis of patients with multiple myeloma
Survival analysis was performed on 121 patients with multiple myeloma with prognostic information by Kaplan-Meier method, and according to the above step 3, the patients were divided into two groups (AZGP 1 high-expression group and AZGP1 low-expression group) according to the relative expression amount of AZGP1, and it was found that 60 month progression-free survival (PFS) was 0% and 18.0% in the AZGP1 high-expression group (n=83) patient and the AZGP1 low-expression group (n=38) patient, respectively, and that the AZGP1 high-expression group patient had a shorter progression-free survival time (median 19.5 months vs.27.7 months, p=0.0064) than the AZGP1 low-expression group patient (fig. 4).
Since there are many factors affecting MM prognosis, to further clarify the effect of AZGP1 on prognosis, the following factors were included in the multifactor analysis: ISS stage at initial diagnosis (stage I-II vs. stage III), bone marrow plasma cell proportion at initial diagnosis (more than or equal to 47% vs. < 47%), blood calcium level at initial diagnosis (more than 2.65mmol/L vs.. Less than or equal to 2.65 mmol/L), hematopoietic stem cell transplantation (whether vs. no). The results show that high AZGP1 expression and ISS stage iii are risk factors for progression free survival (table 3).
TABLE 3 multifactorial analysis of Progression Free Survival (PFS) in adult multiple myeloma patients
End of the year Risk ratio (95% confidence interval) P value
AZGP1: low expression vs. high expression 0.582(0.346-0.979) 0.041*
ISS staging: phase I-II vs. III 0.611(0.377-0.992) 0.046*
Bone marrow plasma cell ratio: more than or equal to 47 percent vs.<47％ 1.307(0.836-2.044) 0.240
Blood calcium level: more than 2.65vs. 2.65 (mmol/L) 1.101(0.603-2.012) 0.754
Receiving hematopoietic stem cell transplantation: is vs. no 0.733(0.405-1.326) 0.304
The present application is described in detail above. It will be apparent to those skilled in the art that the present application can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the application and without undue experimentation. While the application has been described with respect to specific embodiments, it will be appreciated that the application may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the application following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the application pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.

Claims (10)

1. Use of a substance for detecting the expression level of AZGP1 in any one of the following (a 1) to (a 6):
(a1) Preparing a product for diagnosing or aiding in diagnosing multiple myeloma;
(a2) Preparing a product for screening or assisting in screening patients with multiple myeloma;
(a3) Preparing a product for assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
(a4) Preparing a product for assessing or aiding in assessing the progression-free survival of a patient with multiple myeloma;
(a5) Preparing a product for assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(a6) Products are prepared for or aiding in the assessment of progression of disease in patients with multiple myeloma.
2. Use of a substance for detecting the expression level of AZGP1 in any one of the following (b 1) to (b 6):
(b1) Diagnosing or aiding in the diagnosis of multiple myeloma;
(b2) Screening or aiding in screening patients with multiple myeloma;
(b3) Assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
(b4) Assessing or aiding in assessing progression-free survival of a patient with multiple myeloma;
(b5) Assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(b6) The disease course of the patients with the multiple myeloma is evaluated or assisted in evaluation.
3. Use according to claim 1 or 2, characterized in that: the substances for detecting the AZGP1 expression quantity comprise reagents and/or instruments for detecting the AZGP1 expression quantity in bone marrow mononuclear cells.
4. A use according to any one of claims 1-3, characterized in that: the reagent for detecting the AZGP1 expression level in the bone marrow mononuclear cells comprises a specific amplification primer and/or a probe for detecting the mRNA of the AZGP1 gene.
5. The use according to claim 4, characterized in that: the specific amplification primer consists of a single-stranded DNA molecule shown as SEQ ID No.1 and a single-stranded DNA molecule shown as SEQ ID No. 2;
the probe is a single-stranded DNA molecule shown as SEQ ID No. 3.
6. A kit comprising a substance for detecting the expression level of AZGP 1; the function of the kit is any one of the following (c 1) - (c 6):
(c1) Diagnosing or aiding in the diagnosis of multiple myeloma;
(c2) Screening or aiding in screening patients with multiple myeloma;
(c3) Assessing or aiding in assessing the risk of prognosis of a patient with multiple myeloma;
(c4) Assessing or aiding in assessing progression-free survival of a patient with multiple myeloma;
(c5) Assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(c6) The disease course of the patients with the multiple myeloma is evaluated or assisted in evaluation.
7. The kit of claim 6, wherein: the substances for detecting the AZGP1 expression quantity comprise reagents and/or instruments for detecting the AZGP1 expression quantity in bone marrow mononuclear cells.
8. The kit according to claim 6 or 7, wherein: the reagent for detecting the AZGP1 expression level in the bone marrow mononuclear cells comprises a specific amplification primer and/or a probe and a probe for detecting the mRNA of the AZGP1 gene.
9. The kit of claim 8, wherein: the specific amplification primer consists of a single-stranded DNA molecule shown in SEQ ID No.1 and a single-stranded DNA molecule shown in SEQ ID No. 2;
the probe is a single-stranded DNA molecule shown as SEQ ID No. 3.
Use of azgp1 as a biomarker in any one of the following (a 1) - (a 6):
(a1) Preparing a product for diagnosing or aiding in diagnosing multiple myeloma;
(a2) Preparing a product for screening or assisting in screening patients with multiple myeloma;
(a3) Preparing a product for assessing or aiding in assessing prognosis of a patient with multiple myeloma;
(a4) Preparing a product for assessing or aiding in assessing the progression-free survival of a patient with multiple myeloma;
(a5) Preparing a product for assessing or aiding in assessing the progression free survival of a patient with multiple myeloma;
(a6) Products are prepared for or aiding in the assessment of progression of disease in patients with multiple myeloma.
CN202311089468.8A 2023-08-28 2023-08-28 Application of AZGP1 as prognosis risk marker for patients with multiple myeloma Pending CN117089618A (en)

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