CN111443210A - Early diagnosis and treatment biomarker for alveolar echinococcosis and application thereof - Google Patents

Abstract

The invention discloses an application of a substance for detecting target proteins in preparation of products for diagnosing or assisting in diagnosing alveolar echinococcosis, wherein the target proteins are (a1) or (a2) or (A3) (a1) any two of A L DH1A1 protein, TAG L N2 protein and F L N L protein, (a L) A L DH1A L protein, TAG L N L protein and F L N L protein, (a L) A L protein or TAG L N L protein or F L N L protein, the inventor of the invention uses proteome correlation analysis of liver and plasma to detect potential candidate biological AE markers in patients, and the result of the potential AE candidate biological markers in the patients can be used as early diagnosis indexes of AE patients and can be used as early diagnosis candidates for AE patients.

Description

Early diagnosis and treatment biomarker for alveolar echinococcosis and application thereof

Technical Field

The invention relates to a biomarker for early diagnosis and treatment of alveolar echinococcosis and application thereof.

Background

Alveolar echinococcosis is a parasitic disease caused by alveolar Echinococcus (Echinococcus multilocularis), which is also called echinococcosis multicavicularis. The adult echinococcus vesiculosus is similar to the echinococcus granulosus, but the polypide is shorter (1.4-3.4 mm). Unlike echinococcus granulosus, the echinococcus granulosus does not form large vesicles, but forms sponge-like vesicles. The vesicles grow faster, the ascomycetes are exogenic, and the number of metacercaria is also less. The alveolar miracidium is mainly parasitic in the liver and occasionally occurs in the lung and the brain, and the pathological changes and the consequences are more serious than those of echinococcosis granulosa. Macroscopic lesions are usually in the form of single macro-blocks, sometimes nodular, or both. The cyst is usually formed by assembling numerous small vesicles, such as sponge-shaped or honeycomb-shaped vesicles, which are gray in appearance, hard in quality and unclear with surrounding tissues. The vesicle contents are bean curd residue-like larva debris or opaque thin liquid. If the denaturation, necrosis or dissolution occurs, the gel-like liquid is formed. If secondary infection occurs, the abscess is exactly the same. The outer periphery of the vesicular cyst has no complete fibrous capsule, and the growth mode of the vesicular cyst is mainly that the cyst grows outwards to generate a plurality of ascomycetes. The vesicle can infiltrate into surrounding tissues like cancer and can invade blood vessels or lymphatic vessels and spread to the lung, brain, kidney, heart and the like.

Alveolar echinococcosis is one of the most lethal helminth infections and has a high incidence in many parts of the world, including northwest, middle asia, western europe and middle east of china. Moreover, in China, Xinjiang, Qinghai, Tibet and other pasturing areas are more high-incidence areas, especially in the three-river source areas of Qinghai province. More importantly, alveolar echinococcus is mainly present in the liver and can spread to adjacent organs and tissues, sometimes metastasize to distant organs including the lungs and brain. Combination of radical surgery and albendazole (10 mg/kg twice daily) in combination for two years is a strongly recommended treatment in operable patients. Although the number of patients with alveolar echinococcosis is limited, it also imposes a huge financial burden on the patients, and in europe, each patient is loaded with 108,762 euro on average, which is also the main burden of the community in the epidemic area of china. Indeed, the diagnosis of alveolar echinococcosis depends to a large extent on a combination of imaging and serological examinations, since most patients only describe few or vague clinical symptoms and signs. Therefore, the new detection method and early diagnosis have extremely important significance and effect on preventing and treating the alveolar echinococcosis.

Molecular biological methods demonstrate that alveolar echinococcosis can induce T cell failure by the inhibitory receptor TIGIT (T cell immunoreceptor with immunoglobulin and tyrosine-based immunoreceptor inhibitory motif), which is an important new immunotherapy and also a potential biomarker for diagnosing alveolar echinococcosis, and can reverse functional damage to T cells while blocking checkpoints, similarly miRNA microarray analysis indicates that various has-mirnas including miR-483-3p in alveolar echinococcosis tissue are significantly increased and confirm that these mirnas are associated with cellular immune responses (T cell proliferation), further experiments indicate that miR-483-3p can be a potential marker for diagnosing alveolar echinococcosis, I L-5 and I L-23 in alveolar echinococcosis patients are increased in both liver and plasma, and that these are considered to be useful as diagnostic markers for early diagnosis of alveolar echinococcosis (e.g. diagnostic imaging of acute liver diseases) and diagnostic imaging of acute alveolar diabetes, and diagnostic imaging of acute myelodysplasia (e.e. PET-x) using positron emission tomography (MRI), and MRI).

Disclosure of Invention

The invention aims to provide a biomarker for early diagnosis and treatment of alveolar echinococcosis and application thereof.

In a first aspect, the invention firstly protects the application of a substance for detecting a target protein in preparing a product for diagnosing or assisting in diagnosing alveolar echinococcosis; the target protein is (a1) or (a2) or (a3) as follows:

(a1) a L DH1a1 protein and TAG L N2 protein and F L N α protein;

(a2) any two proteins of A L DH1A1 protein, TAG L N2 protein and F L N α protein;

(a3) a L DH1A1 protein or TAG L N2 protein or F L N α protein.

In a second aspect, the invention provides the use of a substance for detecting a target protein and a vector carrying a diagnostic method for the manufacture of a product for diagnosing or aiding in the diagnosis of alveolar echinococcosis; the target protein is (a1) or (a2) or (a3) as follows:

(a1) a L DH1a1 protein and TAG L N2 protein and F L N α protein;

(a2) any two proteins of A L DH1A1 protein, TAG L N2 protein and F L N α protein;

(a3) a L DH1A1 protein or TAG L N2 protein or F L N α protein;

the diagnosis method comprises the following steps: respectively detecting the expression quantity and/or activity of the target protein in the sample of the person to be detected and the control sample, and comparing the detection values; if the expression amount and/or activity value of the target protein in the sample of the subject is larger than the expression amount and/or activity value of the target protein in the control sample, the subject is or is selected as a patient with alveolar echinococcosis; the control sample is a sample of a healthy subject not suffering from alveolar echinococcosis.

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

The sample may specifically be a plasma sample.

In a third aspect, the invention protects the use of a vector describing the above diagnostic method in the manufacture of a product; the product is used for detecting or assisting in detecting the alveolar echinococcosis.

In a fourth aspect, the invention provides a protected product comprising a substance for detecting a target protein; the product is used for diagnosing or assisting in diagnosing the alveolar echinococcosis; the target protein is (a1) or (a2) or (a3) as follows:

(a1) a L DH1a1 protein and TAG L N2 protein and F L N α protein;

(a2) any two proteins of A L DH1A1 protein, TAG L N2 protein and F L N α protein;

(a3) a L DH1A1 protein or TAG L N2 protein or F L N α protein.

Further, the product also comprises a carrier which is recorded with the diagnosis method.

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

(A1) a substance for detecting a target protein; the target protein is (a1) or (a2) or (a3) as follows:

(a1) a L DH1a1 protein and TAG L N2 protein and F L N α protein;

(a2) any two proteins of A L DH1A1 protein, TAG L N2 protein and F L N α protein;

(a3) a L DH1A1 protein or TAG L N2 protein or F L N α 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 (A1) the detected expression quantity and/or activity value of the target protein in the sample of the person to be detected;

the data comparison module is used for comparing the target protein expression quantity and/or activity value in the sample of the testee with the control value; the control value is the expression level and/or activity of the target protein in a sample of a healthy subject not suffering from alveolar echinococcosis;

the conclusion output module is used for outputting the conclusion according to the following standards: if the expression level and/or activity value of the target protein in the sample of the subject is greater than the control value, the subject is or is selected as a patient with alveolar echinococcosis.

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

The sample may specifically be a plasma sample.

The substance for detecting the expression level and/or activity of the target protein further comprises an E L ISA kit, an E L0 ISA kit for detecting the A L DH1A1 protein can be specifically Shanghai Jianella Biotech Co., Ltd., product No. J L19746, an E L ISA kit for detecting TAG L N2 can be specifically Shanghai Jianella Biotech Co., Ltd., product No. J L47102, an E L ISA kit for detecting F L N α can be specifically Shanghai Jianella Biotech Co., Ltd., product No. J L19895.

In a sixth aspect, the invention protects the application of the target protein as a marker in diagnosing or assisting in diagnosing the alveolar echinococcosis; the target protein is (a1) or (a2) or (a3) as follows:

(a1) a L DH1a1 protein and TAG L N2 protein and F L N α protein;

(a2) any two proteins of A L DH1A1 protein, TAG L N2 protein and F L N α protein;

(a3) a L DH1A1 protein or TAG L N2 protein or F L N α protein.

Any one of the aforementioned alveolar echinococcosis may specifically be human alveolar echinococcosis.

The results show that A L DH1A1, TAG L N2 and F L N α are possible early prediction indexes of the Pachybotrys japonica patients and can be used as candidate diagnosis indexes of early AE patients.

Drawings

FIG. 1 shows the results of pathological evaluation of a sample.

FIG. 2 is the global distribution of proteome data.

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

Figure 4 is an enrichment assay for the differential protein GSEA.

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

FIG. 6 shows the results of bioinformatic analysis of common differential proteins in tissues and blood samples.

FIG. 7 shows the detection of marker protein expression and ROC curve.

Detailed Description

The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. All data in the examples below were analyzed using one-way analysis of variance and shown as mean ± SEM, p-values <0.05 were considered statistically significant, and Principal Component Analysis (PCA) and Receiver Operating Characteristic (ROC) curves were performed using SIMCA 14.

Example 1 proteomics analysis

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

First, pathological evaluation

Excised livers from participants with Alveolar Echinococcosis (AE) were divided into three groups, (1) Normal tissue of the liver (Normal), (2) diseased tissue of the liver (L esion), (3) marginal portion of diseased tissue (Edge), located between Normal and diseased tissue histological examination showed no alveolar echinococcosis invasion in the Normal part (FIGS. 1A, D), but tissue lesions were found in the hepatic fibrous tissue (FIGS. 1B, C, D).

Second, protein expression profiles in liver tissues

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

The sample is set as follows, Normal tissues (Normal) of the liver of five alveolar echinococcosis patients are divided into Normal 1-Normal 5, Normal 1-Normal 5 are mixed into Normal C in equal amount, diseased tissues (L) of the liver of five alveolar echinococcosis patients are divided into L esion 1-L esion5, L esion 1-L esion5 are mixed into L esion C in equal amount, marginal parts (Edge) of the diseased tissues of five alveolar echinococcosis patients are divided into Edge 1-Edge 5, and Edge 1-Edge 5 are mixed into Edge C in equal amount.

1. Protein extraction and digestion

(1) 0.1g of liver tissue sample was extracted by adding extraction buffer [ 0.1% SDS (Affymetrix/ThermoFisher Scientific, USA), 100mM Tris/HCl (Affymetrix/Thermo Fisher Scientific, USA) pH 7.6, 10mM DTT (Affymetrix/Thermo Fisher Scientific, USA) ].

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

(3) After completion of step (2), the extract was vortexed at room temperature for 1 minute, centrifuged at 14,000g at 4 ℃ for 20 minutes, 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 labeled 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 reduced with 120. mu. L buffer (10mM DTT, 8M urea, 100mM TEAB, pH 8) at 60 ℃ for 1h, then alkylated with iodoacetamide (IAA; Sigma-Aldrich/Merck, USA, 50mM) for 40min in the dark, and then digested with trypsin according to the FASP method.

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

2. L C-MS/MS analysis

The sample obtained in step 1 was analyzed by Q-active mass spectrometry using nano UP L C (EASY-n L C1200) (Thermo Finnigan, Waltham, MA.) phase inversion column (100 μm, ID × 15cm, Reprosil-Pur120C 18-AQ, 1.9 μm, Dr. Math) for separation₂O, containing 0.1% FA, 2% ACN) and phase B (80% ACN, 0.1% FA). the separation of the samples is carried out at a flow rate of 300n L/min with a gradient of 120min, the mobile phase gradient being 95% mobile phase A and 5% mobile phase B for 5 min, the mobile phase B being from 8% to 30%, 60 min and the mobile phase B being from 30%To 50%, 25 minutes; mobile phase B from 50% to 80%, 5 minutes; the mobile phase B is kept for 15 minutes at 80 percent; mobile phase B from 80% to 5%, 2 minutes; mobile phase B was held for 5%, 8 minutes; the total time is 120 min.

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

3. MaxQuant database search

Protein identification and L FQ. analysis using the enzymatic rule of trypsin/P using MaxQuant (version 1.6.3.4), alignment of Uniprot _ organization _2016_09) protein identification and L FQ. enzymatic rules of cleavage with at most two deletions using trypsin/P the tolerance of the main search peptide was set to 50ppm, the ion trap MS/MS match tolerance was set to 0.5da, the level of peptide match to the profile was set to 0.01FDR, the Andromeda score for the modified peptide was 40, the protein FDR was set to 0.01 and estimated using the reverse search sequence the standard set for MaxQuant was used to perform unlabeled quantitation, the potential contaminants and reverse sequences were removed and it was considered successful to identify proteins with q values <0.05 in each set of MS data, not less than two peptides identified in at least two biological replicates was finally included in the subsequent analysis.

4. Bioinformatics analysis

Gene Ontology (GO) enrichment analysis was performed online using the Blast2GO software program (website: http:// www.geneontology.org) versus Uniprot. Pathway analysis was performed using "Kyoto encyclopedia of genomics" (KEGG) (http:// www.genome.jp/KEGG/mapper. html). Furthermore, interaction analysis of the differential protein was performed using STRING (http:// STRING-db. org /), and visual analysis of protein-protein interaction (PPI) was performed using ClueGo. Pheatmap (version 1.0.12) and ggplot2 (version 3.0.0) of the R software (version 3.5.0) were used to visualize the heat map and bubble map, respectively. Gene Set Enrichment Analysis (GSEA) was performed according to the methods (literature: Subramanian A, Tamayo P, Mootha VK, et al. Gene set expression analysis: a knowledge-based approach for interpretation of genes-with expression profiles, proceedings of the National Academy of sciences of the United States of America.2005; 102: 15545. 15550. Ang. YS, Rivas RN, Ribeiro AJS, et al. area Model of GATA4 Mutation recovery transcription operator Cooperation in Human genetics. cell.2016; 167: 1734. e 1722.).

5. Results

The protein expression profiles in the liver group are shown in FIG. 2A, with the protein expression profile of each group being significantly different, particularly the normal protein, with the profile being reversed from that of the other two groups, for the Edge group the protein profile is more similar to that of the L esion group, where a PCA scoring plot is also performed to directly distinguish the expression profiles, two different portions are shown in the plot, indicating that AE changes the protein expression profile of the lesion compared to the normal group (FIG. 2B). Again, both the heatmap and PCA analysis show similar results (FIGS. 2A, B).

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

The results of GO enrichment analysis 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, α -amino acid catabolic processes, α -amino acid catabolic process preparation, monocarboxylic acid catabolic processes, fatty acid β -oxidation, fatty acid catabolic processes, fatty acid oxidation and lipid oxidation.

GO enrichment analysis was further performed on the DEP shared between lesion/normal and lesion/margin and between lesion/normal and margin/normal. First, DEP shared between diseased/normal and diseased/borderline is significantly enriched in coenzyme biosynthesis processes, carboxylic acid catabolism processes, long chain fatty acid catabolism processes and organic acid catabolism processes. Within the cellular component, DEPs are mainly enriched in Fibrogel-1 particles, microbodies and peroxisomes. Finally, within molecular function, DEP is mainly rich in oxidoreductase activity and serine hydrolases. Then, for the shared DEP between diseased/normal and borderline/normal, proteins associated with the biological process of phagocytosis will be significantly enriched. Within the cellular component, however, DEP is mainly rich in structural components of the extracellular matrix and electrons. Finally, in the molecular function range, DEPs are mainly enriched in aniline-philic blue particles, aniline-philic blue particle membranes and primary lysosomes.

GSEA analysis was performed using the GO gene set for specific DEPs in the lesion/normal group. Partial results are shown in FIG. 4. In the lesion/normal group, genes involved in 7 biological processes are associated with genes upregulated in the normal group, including micronucleus ribonucleoprotein complex, glycosaminoglycan metabolic process, negative regulation of protein secretion, positive regulation of reactive oxygen species metabolic process, protein phosphatase binding and regulation of nitric oxide biosynthetic process. These results indicate that specific DEP may be a candidate biomarker in further treatment of AE.

Pathway analysis of specific DEPs in the lesion/normal group shows that the enriched pathways include mainly arginine biosynthesis, oxidative phosphorylation, citrate cycle (TCA cycle), heterologous organism metabolism by cytochrome P450 and degradation by valine, leucine and isoleucine (also known as BCAA), etc., indicating that this development of AE is a common role of multiple DEPs involved in various pathways.

The development of AE is the result of multiple DEPS interactions and therefore PPI analysis was used to determine interacting proteins the results showed that there were 9 patterns of cluster interactions, mainly including cluster 1, ATP5H interacting with UQCRC2, ATP5J, ATP5A1, ATP5O, ATP5D, NDUFB10, COX5 10, COX 710, NDUFA 10, cluster 2, ARG 10 interacting with VAT 10, ECH 10, PRDX 10, GSTK 10, ANXA 10, EPHX 10, C10, AGXT, DYNC1H 10, ORM 10, HMGC 10, PYGB and CAT, cluster 3, A10 DH6A 10 interacting with ACSS 10, ACAA 10, DBT, HAB, PCCA and PCCB, cluster 4, MDH 10 interacting with SDFH, SDHA, SUC 10 and SUC 10.

Third, protein expression profiling in plasma

Plasma samples from five clinically confirmed participants (PP) with Alveolar Echinococcosis (AE) and from five Normal Participants (NP) were used, all plasma samples were centrifuged at 3000rpm for 20 minutes, and the supernatants were subjected to proteomic analysis with reference to the above method. The results are shown in FIG. 5. 297 DEPs were successfully identified in the PP/NP group, with four groups sharing 2 proteins, three groups except the lesion/margin group sharing 22 proteins, and PP/NP and lesion/normal groups sharing 34. However, co-expression proteomic analysis between PP/NP and lesion/normal groups would be the most likely approach to screening potential candidate biomarkers, particularly from proteins common to both groups. Thus, a common DEP (total of 101 DEPs) determined between the lesion/normal and PP/NP groups was selected for further analysis.

GO enrichment analysis was performed to identify specific functions of the consensus DEP. The shared DEP is rich in proteins associated with biological processes, including regulation of coagulation response, lipoprotein particle assembly, positive regulation of actin filaments, regulation of growth factors, regulation of phospholipase activity, glycosaminoglycan metabolic processes, and negative regulation of 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, DEP was mainly enriched in lipoprotein particles within the cellular component (fig. 6).

Meanwhile, pathway analysis was performed to identify specific functions of the common 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 HIF-1 signaling pathways, indicating that the development of AE is a consequence of the role of various pathways.

Combining the above results, proteins between PP/NP and lesion/normal groups, including A L DH1A1, TAG L N2, F L N α 1A and Igk, were screened as candidate markers.

Example 2 validation of markers

The samples of this example were plasma samples from fifty-six clinically confirmed AE patients and fifty-five healthy controls (not suffering from alveolar 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.

Plasma samples were incubated at room temperature for 20 minutes, then centrifuged at 3000rpm for 20 minutes, the supernatant was transferred to a new tube, and the samples were stored at-80 ℃ for further analysis of the E L ISA.

The E L ISA assay was performed using a commercial kit according to the instructions.

The A L DH1A1 detection kit is purchased from Shanghai Jianglai biological science and technology Limited, with the code of J L19746.

The TAG L N2 detection kit is purchased from Shanghai Jianglai Biotechnology GmbH, with the product number of J L47102.

The F L N α detection kit is purchased from Shanghai Jianglai Biotechnology GmbH, with the product number of J L19895.

The ApoH detection kit is purchased from Wuhan Gene American biotechnology limited company, and has the following product number: JYM0783 Hu.

The CORO1A test kit is purchased from Shanghai Jianglai Biotech limited, cat # J L47869.

The Igk test kit is available from Shanghai Jianglai Biotech Co., Ltd, cat # J L47834.

The results are shown in FIG. 7. the results show that in AE participants, the activity of four proteins including A L DH1A1, TAG L N2, F L N α and ApoH is significantly increased, however, the activity of CORO1A and Igk is not different in both participants, meanwhile, the area under the curve (AUC) of the receiver operating characteristic curve (ROC) proves that all the proteins have high prediction and discrimination capabilities, especially A L DH1A1, TAG L N2 and F L N α, and the area under the ROC line of the three proteins is 84.09%, 82.95% and 77.65%, respectively, and the area under the ROC line of the three proteins is 90.15% (combination), the diagnosis sensitivity is 83.3%, the specificity is 90.9%, the positive predictive value is 91.7% and the negative predictive value is 86%.

The above results indicate that three proteins, a L DH1a1, TAG L N2 and F L N α, are likely candidate evaluation indicators for developing AE.

Claims (10)

1. The use of a substance for detecting a target protein for the preparation of a product for the diagnosis or the auxiliary diagnosis of alveolar echinococcosis; the target protein is (a1) or (a2) or (a3) as follows:

(a1) a L DH1a1 protein and TAG L N2 protein and F L N α protein;

(a2) any two proteins of A L DH1A1 protein, TAG L N2 protein and F L N α protein;

(a3) a L DH1A1 protein or TAG L N2 protein or F L N α protein.

2. The use of a substance for detecting a target protein and a vector carrying a diagnostic method for the preparation of a product for diagnosing or aiding in the diagnosis of alveolar echinococcosis; the target protein is (a1) or (a2) or (a3) as follows:

(a1) a L DH1a1 protein and TAG L N2 protein and F L N α protein;

(a2) any two proteins of A L DH1A1 protein, TAG L N2 protein and F L N α protein;

(a3) a L DH1A1 protein or TAG L N2 protein or F L N α protein;

the diagnosis method comprises the following steps: respectively detecting the expression quantity and/or activity of the target protein in the sample of the person to be detected and the control sample, and comparing the detection values; if the expression amount and/or activity value of the target protein in the sample of the subject is larger than the expression amount and/or activity value of the target protein in the control sample, the subject is or is selected as a patient with alveolar echinococcosis; the control sample is a sample of a healthy subject not suffering from alveolar echinococcosis.

3. Use of a vector as claimed in claim 2 describing a diagnostic method in the manufacture of a product; the product is used for detecting or assisting in detecting the alveolar echinococcosis.

4. A product comprising a substance for detecting a protein of interest; the product is used for diagnosing or assisting in diagnosing the alveolar echinococcosis; the target protein is (a1) or (a2) or (a3) as follows:

(a1) a L DH1a1 protein and TAG L N2 protein and F L N α protein;

(a2) any two proteins of A L DH1A1 protein, TAG L N2 protein and F L N α protein;

(a3) a L DH1A1 protein or TAG L N2 protein or F L N α protein.

5. The product of claim 4, wherein: the product further comprising a vector as defined in claim 3, which is loaded with a diagnostic method.

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

(A1) a substance for detecting a target protein; the target protein is (a1) or (a2) or (a3) as follows:

(a1) a L DH1a1 protein and TAG L N2 protein and F L N α protein;

(a2) any two proteins of A L DH1A1 protein, TAG L N2 protein and F L N α protein;

(a3) a L DH1A1 protein or TAG L N2 protein or F L N α 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 (A1) the detected expression quantity and/or activity value of the target protein in the sample of the person to be detected;

the data comparison module is used for comparing the target protein expression quantity and/or activity value in the sample of the testee with the control value; the control value is the expression level and/or activity of the target protein in a sample of a healthy subject not suffering from alveolar echinococcosis;

the conclusion output module is used for outputting the conclusion according to the following standards: if the expression level and/or activity value of the target protein in the sample of the subject is greater than the control value, the subject is or is selected as a patient with alveolar echinococcosis.

7. The use or product or system of any of claims 1 to 6, wherein: the substance for detecting the target protein is a substance for detecting the expression amount and/or activity of the target protein.

8. The use or product or system of claim 7, wherein: the substance for detecting the expression level and/or activity of the target protein is a substance for detecting the expression level and/or activity of the target protein in blood plasma.

9. The use, product or system of claim 8 wherein the substance for detecting the expression level and/or activity of a target protein in plasma comprises E L ISA kit.

10. The application of the target protein as a marker in diagnosing or assisting in diagnosing the alveolar echinococcosis; the target protein is (a1) or (a2) or (a3) as follows:

(a1) a L DH1a1 protein and TAG L N2 protein and F L N α protein;

(a2) any two proteins of A L DH1A1 protein, TAG L N2 protein and F L N α protein;

(a3) a L DH1A1 protein or TAG L N2 protein or F L N α protein.

Images