CN111443210B - Bubble type echinococcosis early diagnosis and treatment biomarker and application thereof - Google Patents

Abstract

The invention discloses a bubble-type echinococcosis early diagnosis and treatment biomarker and application thereof. The invention protects the application of a substance for detecting target proteins in preparing a product for diagnosing or assisting in diagnosing the echinococcosis; the target protein is (a 1) or (a 2) or (a 3) as follows: (A1) ALDH1A1 protein and TAGLN2 protein and flnα protein; (a2) Any two proteins of ALDH1A1 protein, TAGLN2 protein, and flnα protein; (a 3) ALDH1A1 protein or TAGLN2 protein or FLNα protein. The inventors of the present invention used proteomic association analysis of liver and plasma to detect potential candidate biomarkers in AE patients. The results indicate that ALDH1A1, TAGLN2 and FLNα are possible early predictive indicators of AE patients and can be used as candidate diagnostic indicators of early AE patients.

Description

Bubble type echinococcosis early diagnosis and treatment biomarker and application thereof

Technical Field

The invention relates to a bubble-type echinococcosis early diagnosis and treatment biomarker and application thereof.

Background

The echinococcosis is a parasitic disease caused by echinococcosis bullosa (Echinococcus multilocularis), also called echinococcosis multiforme. The adults of echinococcus bubbly are similar to echinococcus granulosus, but the bodies are shorter (1.4-3.4 mm). Unlike echinococci granulosa, metacercaria does not form large vesicles, but rather spongy vesicles. The vesicle grows faster, the ascus is exogenous, and the number of the metacercaria is small. The metacercaria is mainly parasitic in the liver, even in the lung and brain, and the lesions and the consequences are heavier than those of echinococcosis granulosa. The lesions are typically in the form of individual giant blocks, sometimes nodular, or both. The vesicle is usually formed by assembling numerous small vesicles, such as sponges or honeycombs, and the vesicles are gray in appearance, hard in quality and not clearly demarcated with surrounding tissues. The vesicle content is bean curd residue-like cercaria body scraps or opaque thin liquid. If denaturation, necrosis or dissolution occurs, the gel-like liquid is formed. Such as secondary infections, are similar to abscesses. The periphery of the cyst is not provided with an intact fiber envelope, and the growth mode of the cyst is mainly to bud outside the cyst to generate a plurality of asca. The vesicles can infiltrate the surrounding tissues like cancer and invade blood vessels or lymphatic vessels and spread to the lung, brain, kidneys, heart and the like.

Paulownia is one of the most deadly worm infections, and there is a high incidence in many parts of the world, including northwest, middle asia, western europe and the middle east of china. Moreover, in China, xinjiang, qinghai, tibet and other pastures are more highly developed areas, particularly the Sanjiang source area of Qinghai province. More importantly, echinococci are present primarily in the liver and can spread to adjacent organs and tissues and sometimes metastasize to distant organs including the lungs and brain. Co-treatment with combination of curative surgery and albendazole (twice daily dose 10 mg/kg) in operable patients for two years is a strongly recommended treatment. Although the number of echinococcosis patients is limited, this also places a great financial burden on the patients, in europe, each patient is equally burdened with 108,762 euros, which is of course also the main burden of communities in epidemic regions of china. Indeed, diagnosis of Paulownia is largely dependent on a combination of imaging and serological examinations, as most patients only demonstrate minimal or ambiguous clinical symptoms and signs. Therefore, the novel detection method and the early diagnosis have extremely important significance and effect on preventing and treating the bubble-type echinococcosis.

As a major target organ for bleb echinococcosis, several biomarkers have been identified in liver tissue using various novel methods. Molecular biology methods demonstrate that echinococcosis can induce T cell failure via the inhibitory receptor TIGIT (T cell immune receptor with the inhibitory motif domain of immunoglobulin and tyrosine-based immune receptor), whose expression is enhanced in the liver of chronic AE patients, and can reverse functional damage T cells while blocking checkpoints, suggesting TIGIT is an important novel immunotherapy, as well as a potential biomarker for diagnosing echinococcosis in vesicles. Similarly, miRNA microarray analysis showed a significant increase in a variety of has-mirnas including miR-483-3p in bleacher tissues and demonstrated that these mirnas are associated with cellular immune responses (T cell proliferation). Further experiments show that miR-483-3p can be used as a potential marker for diagnosing the hepatic bleb type echinococcosis. IL-5 and IL-23 are elevated in both liver and plasma in patients with vesicular echinococcosis and are also considered markers of pathological metabolic activity in plasma. But these potential markers lack further validation. In addition, imaging diagnostics, including Ultrasound (US), magnetic Resonance Imaging (MRI), fluorodeoxyglucose positron emission tomography (FDG-PET) and Computed Tomography (CT), as well as serological examinations, such as immunoblotting (WB) and enzyme-linked immunosorbent assays (ELISA), are also used as potential methods for diagnosing blister-type echinococcosis, which are expected to provide more data for the detection of the disease process characterization, or to detect effective biomarkers for the prevention and treatment of blister-type echinococcosis. However, even with these advanced imaging techniques, it is still difficult to diagnose early stage Paulownia effectively.

Disclosure of Invention

The invention aims to provide a bubble-type echinococcosis early diagnosis and treatment biomarker and application thereof.

In a first aspect, the invention first provides the use of a substance for detecting a protein of interest for the preparation of a product for the diagnosis or assisted diagnosis of echinococcosis; the target protein is (a 1) or (a 2) or (a 3) as follows:

(a1) ALDH1A1 protein and TAGLN2 protein and flnα protein;

(a2) Any two proteins of ALDH1A1 protein, TAGLN2 protein, and flnα protein;

(a3) ALDH1A1 protein or TAGLN2 protein or flnα protein.

In a second aspect, the invention provides the use of a substance for detecting a protein of interest and a carrier describing a diagnostic method for the preparation of a product for the diagnosis or for the assisted diagnosis of echinococcosis; the target protein is (a 1) or (a 2) or (a 3) as follows:

(a1) ALDH1A1 protein and TAGLN2 protein and flnα protein;

(a2) Any two proteins of ALDH1A1 protein, TAGLN2 protein, and flnα protein;

(a3) ALDH1A1 protein or TAGLN2 protein or flnα protein;

the diagnosis method comprises the following steps: detecting the expression quantity and/or activity of target protein in the sample of the person to be detected and the control sample respectively, and comparing the detected values; if the expression quantity and/or activity value of the target protein in the sample of the tested person is greater than the expression quantity and/or activity value of the target protein in the control sample, the tested person is or is candidate to be a bubble echinococcosis patient; the control sample is a sample of a healthy person who does not have echinococcosis.

The greater than may specifically be a statistically significant greater than.

The sample may in particular be a plasma sample.

In a third aspect, the invention provides the use of a vector carrying the diagnostic method described above for the preparation of a product; the product is used for detecting or assisting in detecting the echinococcosis.

In a fourth aspect, the present invention provides a protective product comprising a substance for detecting a protein of interest; the product is used for diagnosing or assisting in diagnosing the echinococcosis; the target protein is (a 1) or (a 2) or (a 3) as follows:

(a1) ALDH1A1 protein and TAGLN2 protein and flnα protein;

(a2) Any two proteins of ALDH1A1 protein, TAGLN2 protein, and flnα protein;

(a3) ALDH1A1 protein or TAGLN2 protein or flnα protein.

Further, the product also includes a carrier on which the above diagnostic method is recorded.

In a fifth aspect, the present invention protects a system for diagnosing or aiding in the diagnosis of echinococcosis, comprising:

(A1) A substance for detecting a target protein; the target protein is (a 1) or (a 2) or (a 3) as follows:

(a1) ALDH1A1 protein and TAGLN2 protein and flnα protein;

(a2) Any two proteins of ALDH1A1 protein, TAGLN2 protein, and flnα protein;

(a3) ALDH1A1 protein or TAGLN2 protein or flnα protein;

(A2) The device comprises a data input module, a data comparison module and a conclusion output module;

the data input module is used for inputting the target protein expression quantity and/or the activity value in the sample of the to-be-detected person detected by the (A1);

the data comparison module is used for comparing the target protein expression quantity and/or the activity value in the sample of the person to be tested with a control value; the control value is the expression quantity and/or activity of target protein in a healthy subject sample without the echinococcosis;

the conclusion output module is used for outputting a conclusion according to the following standard: if the expression level and/or the activity value of the target protein in the sample of the tested person is greater than the control value, the tested person is or is candidate to be a patient with the echinococcosis.

The greater than may specifically be a statistically significant greater than.

The sample may in particular be a plasma sample.

In the above aspects, the substance for detecting a target protein is a substance for detecting an expression amount and/or activity of the target protein. Further, the substance for detecting the expression amount and/or activity of the target protein is a substance for detecting the expression amount and/or activity of the target protein in plasma. Further, the substance for detecting the expression amount and/or activity of the target protein in the blood plasma comprises an ELISA kit. For example, the ELISA kit for detecting ALDH1A1 protein can be specifically obtained from Shanghai Jiang Lai Biotechnology Inc. under the product of JL19746. For example, the ELISA kit for detecting TAGLN2 can be specifically obtained from Shanghai Jiang Lai Biotechnology Inc. under the product of JL47102. For example, the ELISA kit for detecting FLNα may be specifically manufactured by Shanghai Jiang Lai Biotechnology Inc. under the product of JL19895.

In a sixth aspect, the invention provides the use of a protein of interest as a marker for diagnosis or assisted diagnosis of echinococcosis; the target protein is (a 1) or (a 2) or (a 3) as follows:

(a1) ALDH1A1 protein and TAGLN2 protein and flnα protein;

(a2) Any two proteins of ALDH1A1 protein, TAGLN2 protein, and flnα protein;

(a3) ALDH1A1 protein or TAGLN2 protein or flnα protein.

Any of the above-described vesicular echinococcosis may be specifically human vesicular echinococcosis.

The inventors of the present invention used proteomic correlation analysis of liver and plasma to detect potential candidate biomarkers in patients with echinococcosis. The results show that ALDH1A1, TAGLN2 and FLNα are possible early predictive indicators of patients with echinococcosis, and can be used as candidate diagnostic indicators of early AE patients.

Drawings

Fig. 1 is a sample pathology evaluation result.

FIG. 2 shows the overall distribution of proteome data.

FIG. 3 shows the basic situation of differentially expressed proteins.

FIG. 4 is a differential protein GSEA enrichment analysis.

FIG. 5 shows the results of a combined analysis of a tissue sample and a blood sample.

FIG. 6 shows the results of bioinformatic analysis of differential proteins shared by tissues and blood samples.

FIG. 7 shows marker protein expression assays and ROC curves.

Detailed Description

The following examples facilitate a better understanding of the present invention, but are not intended to limit the same. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the examples described below, unless otherwise specified, were purchased from conventional biochemical reagent stores. All data in the examples below were analyzed using one-way anova and shown as mean ± SEM, p-values <0.05 were considered statistically significant, and Principal Component Analysis (PCA) and subject operating characteristics (ROC) curves were performed using SIMCA 14.

Example 1 proteomic analysis

The samples used in this example were ex vivo liver and plasma samples from five clinically confirmed participants with echinococcosis (AE) and plasma samples from five normal participants (who did not have echinococcosis). The protocol has been approved by the ethics committee of the affiliated hospital at Qinghai university and informed consent has been obtained from each subject.

1. Pathological assessment

Isolated livers from participants with echinococcosis (AE) were divided into three groups: (1) Normal tissue of liver (Normal); (2) liver Lesion tissue (fusion); (3) Edge portions (Edge) of the diseased tissue lie between normal and diseased. Histological examination showed that normal parts were not affected by echinococcosis blepharum (fig. 1a, d), but that histologic lesions were found in liver fibrous tissue (fig. 1b, c, d).

2. Protein expression profiling in liver tissue

Label-free proteomics techniques were used to analyze specific protein expression profiles in three groups of liver tissues.

The sample settings were as follows: the Normal tissues (Normal) of the livers of five patients suffering from bubble-type echinococcosis are classified as Normal1 to Normal5, and equal amounts of Normal1 to Normal5 are mixed and classified as Normal C; the liver Lesion tissues (Lesion) of five bubble-type echinococcosis patients are classified into Lesion1 to Lesion5, and Lesion1 to Lesion5 are equally mixed and classified into Lesion C; edge parts (Edge) of lesion tissues of five patients suffering from the bubble type echinococcosis are classified into Edge1 to Edge5, and equal amounts of Edge1 to Edge5 are mixed and classified into Edge C.

1. Protein extraction and digestion

(1) Liver tissue samples were taken at 0.1g and extracted with extraction buffer [0.1% SDS (Affymetrix/Thermo Fisher Scientific, U.S.) 100mM Tris/HCl (Affymetrix/Thermo Fisher Scientific, U.S.) pH 7.6, 10mM DTT (Affymetrix/Thermo Fisher Scientific, U.S. ].

(2) After completion of step (1), liver samples were homogenized on ice using a homogenizer (T18, germany) for 1 minute.

(3) After the completion of step (2), the extract was vortexed at room temperature for 1 minute, centrifuged at 14,000g for 20 minutes at 4℃and the supernatant was taken.

(4) After completion of step (3), the supernatant sample was heated at 56℃for 30 minutes and then centrifuged at 14,000g at 20℃for 20 minutes. The supernatant was transferred to a new labelling tube, then mixed, aliquoted and stored at-20 ℃ until use.

(5) After quantifying the protein sample prepared in step (4), 0.1mg of protein was taken, reduced with 120. Mu.L of buffer (10 mM DTT,8M urea, 100mM TEAB, pH 8) at 60℃for 1h, then treated with iodoacetamide (IAA; sigma-Aldrich/Merck, USA,50 mM) in the absence of light for 40min, and then digested with trypsin according to the FASP method.

(6) After completion of step 5, the samples were purified using Pierce C18 column (Affymetrix/Thermo Fisher Scientific, USA), dried under vacuum and stored at-80 ℃. For each sample, about 2 μg of peptide was isolated and used for subsequent analysis.

2. LC-MS/MS analysis

The sample obtained in step 1 was analyzed using a Q-exact mass spectrometer coupled with nano UPLC (EASY-nLC 1200) (Thermo Finnigan, waltham, mass.). The separation was carried out using a reverse phase column (100 μm, ID. Times.15 cm, reprosil-Pur 120C 18-AQ,1.9 μm, dr. Math). The mobile phase is A phase (H) ₂ O, 0.1% FA,2% ACN) and phase B (80% ACN,0.1% FA). The separation of the samples was carried out at a flow rate of 300nL/min with a gradient of 120min. The mobile phase gradient is: 95% mobile phase a and 5% mobile phase B for 5 minutes; mobile phase B from 8% to 30%,60 minutes; mobile phase B from 30% to 50%,25 minutes; mobile phase B from 50% to 80%,5 minutes; mobile phase B remained 80% for 15 min; mobile phase B from 80% to 5%,2 minutes; mobile phase B remained 5% for 8 min; the total time was 120min.

Data-dependent acquisition was performed in profile and forward mode using an Orbitrap analyzer with a MS1 resolution of 70,000 (@ 200 m/z) and m/z in the range of 350-1600. For MS2, the resolution is set to 17,500 and has a dynamic first quality. The Automatic Gain Control (AGC) target for MS1 is set to 3.0E+6 for a maximum IT 50 μms, and MS2 is set to 5.0E+4 for a maximum IT 100 μms. HCD breaks the first 20 strongest ions.

3. MaxQuant database search

Protein identification and LFQ were analyzed using MaxQuant (1.6.3.4 version), alignment Uniprot database (uniprot_organization_2016_09). There are at most two deletion cuts using the enzymatic principle of trypsin/P. The tolerance of the main search peptide was set to 50ppm and the ion trap MS/MS matching tolerance was set to 0.5Da. The level of match of the peptide to the profile was set at 0.01FDR and the Andromeda score for the modified peptide was 40. Protein FDR was set to 0.01 and evaluated using the reverse search sequence. The standard set by MaxQuant is used to perform label-free quantification. Potential contaminants and reverse sequences were removed and it was believed that no less than two peptide proteins with q <0.05 were successfully identified in each set of MS data. The proteins identified in at least two biological replicates are ultimately included in subsequent assays. Differentially Expressed Proteins (DEPs) were identified based on 1.5 fold changes with P values < 0.05.

4. Bioinformatics analysis

Gene Ontology (GO) enrichment analysis was performed on Uniprot online using the Blast2GO software program (website: http:// www.geneontology.org). Pathway analysis was performed using the "kyoto encyclopedia of genes and genome" (KEGG) (http:// www.genome.jp/KEGG/mapper. Html). In addition, differential protein interactions were analyzed using STRING (http:// STRING-db. Org /), and protein-protein interactions (PPI) were visually analyzed using ClueGo. Pheatmap (version 1.0.12) and ggplot2 (version 3.0.0) of R software (version 3.5.0) were used to visualize the heat and bubble maps, respectively. Gene enrichment analysis (GSEA) was performed according to the method (literature: subramann A, tamayo P, mootha VK, et al Gene set enrichment analysis: a knowledges-based approach for interpreting genome-width expression profiles. Proceedings of the National Academy of Sciences ofthe United States of America.2005;102: 15545-15550; ang YS, rivas RN, ribeiro AJS, et al disease Model of GATA4 Mutation Reveals Transcription Factor Cooperativity in Human cart. Cell.2016;167:1734-1749e 1722).

5. Results

The protein expression profile in the liver groups is shown in fig. 2A, and the protein expression profile of each group is significantly different, especially normal protein, and the profile is opposite to the other two groups. For Edge group, its protein profile is more similar to that of the Lesion group. Here, PCA scoring was also performed to directly distinguish expression profiles. Two different parts are shown, indicating that AE alters the protein expression profile of the lesions compared to the normal group (fig. 2B). Likewise, both the heat map and PCA analysis showed similar results (fig. 2a, b).

The identification results of DEPs in three groups are shown in FIG. 3, and 2087 DEPs (FIG. 3A) were successfully identified in three groups, of which 1701 DEPs, 884 DEPs and 1184 DEPs belong to lesion/normal (FIG. 3A, 3B), edge/normal (FIG. 3A, 3C) and lesion/edge (FIG. 3A, 3D) groups, respectively. And up-regulating or down-regulating DEP is shown in FIGS. 3B-D.

The GO enrichment analysis results of DEP show that various biological processes related to DEP are significantly enriched, especially organic acid catabolic processes, carboxylic acid catabolic processes, cellular amino acid catabolic processes, alpha-amino acid metabolic processes, alpha-amino acid catabolic process preparation methods, monocarboxylic acid decomposition methods, fatty acid beta-oxidation methods, fatty acid decomposition methods, fatty acid oxidation methods, lipid oxidation methods and the like. The results show that DEPs are significantly enriched for proteins associated with molecular function, mainly carboxylic acid binding, amino acid binding, NAD binding and organic acid binding. Meanwhile, within the cellular component, DEP is localized mainly in peroxisome/peroxisome parts, myelin, mitochondrial membrane parts, microsomes and microsomal parts/cavities.

GO enrichment analysis was further performed on DEP shared between lesions/normal and lesions/edges and DEP shared between lesions/normal and edges/normal. First, DEP shared between lesions/normal and lesions/edges is significantly enriched in coenzyme biosynthesis processes, carboxylic acid catabolic processes, long chain fatty acid metabolic processes and organic acid catabolic processes. Within the cellular component, DEPs are mainly enriched in fiber-rich gel-1 particles, microsomes and peroxisomes. Finally, DEP is mainly rich in oxidoreductase activity and serine hydrolases within the molecular function. Then, for shared DEP between lesions/normal and marginally/normal, proteins associated with phagocytic biological processes will be significantly enriched. However, in cellular components, DEP is mainly rich in structural components of the extracellular matrix and electrons. Finally, within the molecular functional range, DEPs are mainly enriched in the anilinophiles particles, the anilinophiles particle membranes and the primary lysosomes.

GSEA analysis was performed using GO gene sets of specific DEP in the lesion/normal group. The partial results are shown in FIG. 4. In the lesion/normal group, genes involved in 7 biological processes are associated with genes up-regulated in the normal group, including small ribonucleoprotein complexes, glycosaminoglycan metabolic processes, negative regulation of protein secretion, positive regulation of reactive oxygen species metabolic processes, protein phosphatase binding, and regulation of nitric oxide biosynthesis processes. These results indicate that a particular DEP may be a candidate biomarker in AE further treatment.

The results of pathway analysis of specific DEPs in the lesion/normal group show that the enriched pathway mainly includes arginine biosynthesis, oxidative phosphorylation, citrate cycle (TCA cycle), metabolism of heterologous organisms by cytochrome P450 and degradation of valine, leucine and isoleucine (also referred to as BCAA), etc., indicating that this occurrence of developing AE is a co-action of multiple DEPs involved in various pathways. In addition, path analysis was performed for DEPs shared between lesions/normal and lesions/edges, lesions/normal and edges/normal. DEP shared between lesions/normal and lesions/edges is primarily involved in chemokine signaling pathways, leukocyte trans-endothelial migration, NF-. Kappa.B signaling pathways, alzheimer's disease and glutathione metabolism, and the like. However, the DEP shared between lesions/normal and limbus/normal is shown to be different, with proteins localized primarily to regulation of lipolysis in adipocytes, HIF signaling pathways, glycolysis/gluconeogenesis, adhesion, spliceosomes and fatty acid extension.

The development of AE is the result of a variety of DEPS interactions, and thus PPI analysis was used to determine the interacting proteins. The results show that a total of 9 pattern clusters have interactions, including mainly cluster 1, ATP5h interactions with uqrc 2, ATP5J, ATP5A1, ATP5O, ATP5D, NDUFB10, COX5B, COX7C, NDUFA 5; cluster 2, arg1 interacts with VAT1, ECH1, PRDX6, GSTK1, ANXA2, EPHX2, C3, AGXT, DYNC1H1, ORM2, HMGCL, IDH1, PYGB and CAT; in cluster 3, ALDH6A1 interacts with ACSS3, ACAA1, DBT, HADHB, PCCA and PCCB; cluster 4, mdh2 has interactions with FH, SDHA, DLST, SUCLG and SUCLG 2.

3. Protein expression profile in plasma

Plasma samples from five clinically confirmed participants (PP) with echinococcosis (AE) and from five normal participants (no echinococcosis, NP) were used, all plasma samples were centrifuged at 3000rpm for 20 minutes and the supernatants were proteomic analyzed with reference to the above method. The results are shown in FIG. 5. 297 DEPs were successfully identified in the PP/NP group, four of which had 2 total proteins, three groups other than the lesion/limbic group had 22 total proteins, and the PP/NP and lesion/normal groups had 34 total proteins. However, co-expressed proteomic analysis between PP/NP and lesion/normal groups would be the most likely method to screen for potential candidate biomarkers, especially from proteins common to both groups. Thus, a total of 101 DEPs (total) determined between the lesion/normal group and the PP/NP group were selected for further analysis.

GO enrichment analysis was performed to identify specific functions of the shared DEP. The shared DEPs are rich in proteins associated with biological processes, including regulation of the coagulation response, lipoprotein particle assembly, positive regulation of actin filaments, regulation of growth factors, regulation of phospholipase activity, negative regulation of glycosaminoglycan metabolic processes and phosphate (FIG. 6). Within molecular functions, DEP is mainly enriched in protein tyrosine kinase activator activity, peptidoglycan receptor activity and hydrolytic activity (fig. 6). Finally, within the cellular component, DEP is mainly enriched in lipoprotein particles (fig. 6).

At the same time, a path analysis is performed to identify specific functions of the shared DEP. DEPs are mainly enriched in the complement and coagulation cascade, glycolysis/gluconeogenesis, carbon metabolism, metabolic pathways, amino acid biosynthesis, tyrosine metabolism, arginine biosynthesis, fatty acid degradation and the HIF-1 signaling pathway, indicating that AE development is the result of various pathway actions.

In summary, proteins between PP/NP and lesion/normal group, including ALDH1A1, TAGLN2, flnα, apoH, CORO1A and Igk were selected as candidate markers.

Example 2 verification of markers

The samples of this example were plasma samples from fifty-six clinically diagnosed AE patients and fifty-five healthy controls (who did not have echinococcosis). None of the subjects had a history of respiratory or cardiovascular disease, such as chronic obstructive pulmonary disease, asthma, infectious disease, congenital heart disease, or hypertensive heart disease. Informed consent has been obtained from each subject.

The plasma samples were incubated at room temperature for 20 minutes, then centrifuged at 3000rpm for 20 minutes, the supernatant transferred to a fresh tube, and the samples were stored at-80 ℃ for further ELISA analysis.

ELISA assays were performed using commercial kits according to the instructions.

ALDH1A1 detection kit was purchased from Shanghai Jiang Lai Biotechnology Co., ltd., product number: JL19746.

TAGLN2 detection kit was purchased from Shanghai Jiang Lai Biotech Co., ltd., cat: JL47102.

Flnα detection kit was purchased from shanghai Jiang Lai biotechnology limited, cat: JL19895.

ApoH detection kit was purchased from Wuhan Genetime Biotechnology Co., ltd., cat No.: JYM0783Hu.

CORO1A assay kit was purchased from Shanghai Jiang Lai Biotechnology Co., ltd., product number: JL47869.

Igk test kit was purchased from shanghai Jiang Lai biotechnology limited, cat: JL47834.

The detection results are shown in FIG. 7. The results show that the activity of four proteins including ALDH1A1, TAGLN2, FLNα and ApoH was significantly increased in AE participants. However, the activity of CORO1A and Igk was not different in both participants. Meanwhile, the area under the curve (AUC) of the working characteristic curve (ROC) of the subject proves that all the proteins have higher prediction and distinguishing capability, in particular ALDH1A1, TAGLN2 and FLNα, and the areas under the ROC line of the three proteins are 84.09%, 82.95% and 77.65% respectively; the area of the curve under the ROC line combined by the three is 90.15 percent (combination), the diagnosis sensitivity is 83.3 percent, the specificity is 90.9 percent, the positive predictive value is 91.7 percent, and the negative predictive value is 86 percent.

The above results indicate that three proteins, ALDH1A1, TAGLN2 and flnα, are likely candidates for evaluation of the development of AE.

Claims (9)

1. Use of a substance for detecting a protein of interest for the preparation of a product for the diagnosis or assisted diagnosis of echinococcosis; the target protein is as follows (a 1) or (a 2):

(a1) ALDH1A1 protein and TAGLN2 protein and flnα protein;

(a2) ALDH1A1 protein or TAGLN2 protein or flnα protein.

2. Use of a substance for detecting a target protein and a carrier described with a diagnostic method for the preparation of a product for diagnosing or aiding in the diagnosis of echinococcosis; the target protein is as follows (a 1) or (a 2):

(a1) ALDH1A1 protein and TAGLN2 protein and flnα protein;

(a2) ALDH1A1 protein or TAGLN2 protein or flnα protein;

the diagnosis method comprises the following steps: detecting the expression quantity and/or activity of target protein in the sample of the person to be detected and the control sample respectively, and comparing the detection values; if the expression quantity and/or activity value of the target protein in the sample of the tested person is greater than the expression quantity and/or activity value of the target protein in the control sample, the tested person is or is candidate to be a bubble echinococcosis patient; the control sample is a sample of a healthy person who does not have echinococcosis.

3. Use according to claim 1 or 2, characterized in that: the substance for detecting a target protein is a substance for detecting the expression amount and/or activity of the target protein.

4. A use according to claim 3, wherein: the target protein is a target protein in plasma.

5. The use according to claim 4, wherein: the product comprises an ELISA kit.

6. A system for diagnosing or aiding in the diagnosis of echinococcosis bubble comprising:

(A1) A substance for detecting a target protein; the target protein is as follows (a 1) or (a 2):

(a1) ALDH1A1 protein and TAGLN2 protein and flnα protein;

(a2) ALDH1A1 protein or TAGLN2 protein or flnα protein;

(A2) The device comprises a data input module, a data comparison module and a conclusion output module;

the data input module is used for inputting the target protein expression quantity and/or the activity value in the sample of the to-be-detected person detected by the (A1);

the data comparison module is used for comparing the target protein expression quantity and/or the activity value in the sample of the person to be tested with a control value; the control value is the target protein expression quantity and/or activity value in a healthy subject sample without the bubble type echinococcosis;

the conclusion output module is used for outputting a conclusion according to the following standard: if the expression level and/or the activity value of the target protein in the sample of the tested person is greater than the control value, the tested person is or is candidate to be a patient with the echinococcosis.

7. The system of claim 6, wherein: the substance for detecting a target protein is a substance for detecting the expression amount and/or activity of the target protein.

8. The system of claim 7, wherein: the target protein is a target protein in plasma.

9. The system as recited in claim 8, wherein: the substance for detecting the expression amount and/or activity of the target protein in the blood plasma comprises an ELISA kit.