CN117554629B - Marker for differential diagnosis of neurosyphilis and application thereof - Google Patents
Marker for differential diagnosis of neurosyphilis and application thereof Download PDFInfo
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- 101000609413 Homo sapiens Inter-alpha-trypsin inhibitor heavy chain H4 Proteins 0.000 claims abstract description 18
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/26—Infectious diseases, e.g. generalised sepsis
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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Abstract
The invention relates to the technical field of medical detection, in particular to a marker for differential diagnosis of neurosyphilis and application thereof. The invention uses a proteomics combination deep learning method to obtain three biomarkers ITIH4, SERPINA3 and SEMA7A of the neurosyphilis from a cerebrospinal fluid sample, wherein the three biomarkers of the neurosyphilis can distinguish the neurosyphilis patients from non-neurosyphilis patients and the neurosyphilis patients infected by other brain diseases. The present invention demonstrates these three neurosyphilis biomarkers using Elisa and brain histopathology. In addition, the three biomarkers and the neurosyphilis evaluation model are used for predicting suspected neurosyphilis patients which cannot be distinguished clinically, the three biomarkers are found to have higher relevance to the neurosyphilis evaluation model, and the patients with the three biomarkers are found to be diagnosed as neurosyphilis positive in follow-up.
Description
Technical Field
The invention relates to the technical field of medical detection, in particular to a marker for differential diagnosis of neurosyphilis and application thereof.
Background
Early infections of neurosyphilis may be asymptomatic, but can lead to serious organ injury and irreversible dysfunction, even life threatening, and early identification and treatment is therefore critical.
Current diagnostic methods are based primarily on clinical presentation and cerebrospinal fluid testing, but have no accepted diagnostic criteria. Diagnostic criteria in the U.S. centers for disease control and defense divide neurosyphilis into two categories, definitive diagnosis and suspicion, but result in a high proportion of false negatives and misdiagnosis due to the low sensitivity of the test. Therefore, finding biomarkers specific to disease is critical for understanding pathogenesis of neurosyphilis, improving early diagnosis rate, and reducing occurrence of complications.
Although there have been studies using immune factors as diagnostic markers, these markers are also expressed in other neurological diseases. Some studies suggest that uPA, sTREM2, N-acetyl-L-tyrosine and microRNAs may be potential biomarkers, but the sensitivity, specificity and cut-off values of these studies are not consistent due to sample heterogeneity.
In view of this, the present invention has been made.
Disclosure of Invention
In order to solve the technical problems, the invention provides application of a marker or a detection reagent thereof in preparing a product for differential diagnosis of the neurosyphilis, wherein the marker comprises at least one of the following protein markers: ITIH4, SERPINA3, and SEMA7A.
The expression level of any one of the protein markers has obvious difference in the nerve syphilis, the non-nerve syphilis and other brain diseases, can accurately distinguish the nerve syphilis, the non-nerve syphilis and other brain diseases, and has higher specificity and sensitivity. Therefore, the protein marker can be independently used for differential diagnosis of the neurosyphilis.
With respect to the combination of the above markers, it is understood that since any one of the above markers can be used alone for differential diagnosis of neurosyphilis, the combined use thereof can also be used for differential diagnosis of neurosyphilis and may exhibit higher accuracy.
For the markers, the expression levels of ITIH4 and SERPINA3 are up-regulated in the body of a patient with the neurosyphilis compared with the expression levels of the patient without the neurosyphilis; and/or, the expression level of SEMA7A is down-regulated compared to a non-neural syphilis patient.
Preferably, the product is a medicament or a kit.
In a second aspect, the present invention provides the use of a marker or an antagonist thereof in the manufacture of a medicament for the treatment of a neurosyphilis, said marker comprising at least one of the following protein markers: ITIH4, SERPINA3 and SEMA7A; the marker is used for a drug target for treating the nerve syphilis.
In a third aspect, the present invention provides an apparatus for differential diagnosis of a neurosyphilis, the apparatus comprising:
The detection module is used for detecting the content of the marker in the sample to be detected; the markers include at least one of the following protein markers: ITIH4, SERPINA3 and SEMA7A;
the input module is used for acquiring the detection result of the detection module;
the judging module is used for comparing the detection result obtained by the input module with a patient with the nerve syphilis and judging whether the patient is the nerve syphilis or not;
and the output module is used for outputting the diagnosis result.
Preferably, the judging criteria of the judging module include: if the content of ITIH4 and/or SERPINA3 in the sample to be tested is significantly up-regulated compared with that of a non-neurotensin patient, the neurotensin is judged.
Preferably, the judging criteria of the judging module include: and if the SEMA7A content in the sample to be detected is obviously reduced compared with the non-neural syphilis patient, judging that the sample is neural syphilis.
Preferably, the sample to be tested is blood, plasma or serum.
In a fourth aspect, the present invention provides a method of differential diagnosis of a neurosyphilis, the method comprising: detecting the expression quantity of the marker for differential diagnosis of the neurosyphilis in a human body to be diagnosed, and judging whether the neurosyphilis exists or not according to the change condition of the expression quantity.
The basis of the judgment comprises at least one of the following (1) - (3): (1) If the content of ITIH4 in the sample to be detected is obviously up-regulated compared with the non-neural syphilis patient, the neural syphilis is judged. (2) If the content of SERPINA3 in the sample to be tested is obviously up-regulated compared with the non-neural syphilis patient, the sample is judged to be the neural syphilis. (3) And if the SEMA7A content in the sample to be detected is obviously reduced compared with the non-neural syphilis patient, judging that the sample is neural syphilis.
The beneficial effects are that:
Three biomarkers ITIH4, SERPINA3 and SEMA7A of the nervous syphilis were identified from cerebrospinal fluid samples by proteomics methods in this study. These three markers can distinguish between neurosyphilis and non-neurosyphilis patients and are confirmed by Elisa and brain histopathology. The study also combined these three markers with a neurosyphilis evaluation model and confirmed in follow-up that these patients with these three markers were diagnosed as neurosyphilis positive.
Drawings
In order to more clearly illustrate the technical solutions of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be described below.
FIG. 1 is a diagram showing the proteomic profile of cerebrospinal fluid of a patient suffering from a neurosyphilis according to example 1 of the present invention.
FIG. 2 is a graph showing the analysis of the protein co-expression pattern in cerebrospinal fluid of a patient suffering from a neurosyphilis according to example 2 of the present invention.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The invention uses a proteomics combination deep learning method to obtain three biomarkers ITIH4, SERPINA3 and SEMA7A of the neurosyphilis from a cerebrospinal fluid sample, wherein the three biomarkers of the neurosyphilis can distinguish the neurosyphilis patients from non-neurosyphilis patients and the neurosyphilis patients infected by other brain diseases.
The research process of the invention is specifically as follows:
The study divided patients into multiple groups including neurosyphilis group (NS), non-neurosyphilis syphilis group (NNS), etc., and identified diagnostic markers of neurosyphilis by proteomic mass spectrometry and machine learning models of cerebrospinal fluid samples.
Proteins are extracted from cerebrospinal fluid samples by mass spectrometry and data processing to process into polypeptide mixtures. DIA data was analyzed using an Orbitrap Fusion mass spectrometer and Proteome Discoverer and a spectral library of specific samples was generated.
Through five steps of data set preprocessing, feature selection, model training, identification of the most important features and model evaluation, a classifier based on Random Forest (RF) is developed and used for identifying potential biomarkers to classify the neurosyphilis.
Finally, three biomarkers (SEMA 7A, SERPINA3 and ITIH 4) were validated in multiple queues and 115 cerebrospinal fluid samples. Experimental results show that ITIH4 and SERPINA3 expression are up-regulated and SEMA7A expression is down-regulated in the NS group. Cerebrospinal fluid testing of PTNS patients found reduced expression levels of ITIH4 and SERPINA 3. These results indicate that these three proteins can serve as biomarkers for NS diagnosis.
Example 1
Proteomic characteristics and potential biomarkers of cerebrospinal fluid of patients with neurosyphilis
In this example, a total of 40 cerebrospinal fluid samples were collected from 15 neurosyphilis and 25 NNS patients for proteomic testing. Cerebrospinal fluid samples from NNS (n=25), NS (n=15) and other groups of patients were analyzed using quantitative proteomic and bioinformatic analysis methods. Using high resolution mass spectrometry, samples were subjected to data dependent acquisition mode (DDA) library construction, and then each sample was subjected to data independent acquisition mode (DIA) for protein identification and quantification.
The abbreviations referred to in this invention have the following meanings:
NNS: syphilis-nonneural syphilis;
NS: neurosyphilis;
PTNS: post-treatment neurosyphilis;
NIBD: no syphilis but no infectious brain disease;
IBD: there is no syphilis but there is an infectious brain disease.
The present invention uses a non-dependent data acquisition (DIA) strategy, where thousands of proteins are identified, including expressed proteins from the group of neurosyphilis (1749) and NNS (1719). The results showed that protein abundance was associated with the neurosyphilis and NNS group.
Hierarchical Cluster Analysis (HCA) and volcanic image analysis results are shown in fig. 1, panels a and b.
In figure a: proteins expressed differentially (total 358 proteins) in cerebrospinal fluid of NNS (n=25) and NS (n=15) samples (cohort 1). Wherein the red and blue boxes or boxes represent up-and down-regulated proteins of NS compared to NNS.
B, in the graph: biological process analysis of differentially expressed proteins in NS and NNS cerebrospinal fluid was ordered according to log10 p values. The color represents the functional category. The expression levels of selected functional proteins vary, with these proteins being significantly up-regulated (upper right region red font) and down-regulated (lower left region blue font) between NNS and NS samples.
Hierarchical Cluster Analysis (HCA) and volcanic image analysis showed 358 differentially expressed proteins between the Neurosyphilis (NS) and non-neurosyphilis (NNS) groups.
Example 2
To further confirm biomarkers of neurosyphilis, this example constructs a ranking table for protein prioritization based on machine learning model scores and the like, and expands the sample size for RPM and Elisa validation of priority proteins.
The results of the analysis of the protein co-expression pattern in the cerebrospinal fluid of patients with neurosyphilis are shown in FIG. 2. The co-expression pattern of proteins in cluster 1 and cluster 2 modules represents proteins that are up-regulated (cluster 1) and down-regulated (cluster 2) in the NS group compared to the NNS group and recovered in the PTNS group.
The results in FIG. 2 show that the 3 proteins ITIH4, SERPINA3 and SEMA7A can be used as biomarkers for diagnosis of neurosyphilis.
The biomarkers provided in this example can be used in clinical diagnosis of patients with suspected neurosyphilis. These markers can be used to develop in vivo diagnostic products to reduce the occurrence of brain damage in patients with neurosyphilis due to misdiagnosis or missed diagnosis.
The above examples merely represent a few embodiments of the present invention, which facilitate a specific and detailed understanding of the technical solutions of the present invention, but are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention.
Claims (8)
1. Use of a marker for the preparation of a product for the differential diagnosis of neurosyphilis, characterized in that said marker comprises at least one of the following protein markers: ITIH4, SERPINA3, and SEMA7A.
2. The use according to claim 1, wherein the expression level of ITIH4, SERPINA3 is up-regulated in a patient suffering from neurosyphilis compared to a patient suffering from non-neurosyphilis; and/or, the expression level of SEMA7A is down-regulated compared to a non-neural syphilis patient.
3. The use according to claim 1, wherein the markers comprise the following protein markers: ITIH4, SERPINA3 and SEMA7A; the product is a drug or a kit.
4. Use of a marker in the manufacture of a medicament for the treatment of a neurosyphilis, wherein the marker comprises at least one of the following protein markers: ITIH4, SERPINA3 and SEMA7A; the marker is used for a drug target for treating the nerve syphilis.
5. A device for differential diagnosis of a neurosyphilis, said device comprising:
The detection module is used for detecting the content of the marker in the sample to be detected; the markers include at least one of the following protein markers: ITIH4, SERPINA3 and SEMA7A;
the input module is used for acquiring the detection result of the detection module;
the judging module is used for comparing the detection result obtained by the input module with a patient with the nerve syphilis and judging whether the patient is the nerve syphilis or not;
and the output module is used for outputting the diagnosis result.
6. The apparatus for differential diagnosis of neurosyphilis according to claim 5, wherein the judgment criteria of the judgment module comprises: if the content of ITIH4 and/or SERPINA3 in the sample to be tested is significantly up-regulated compared with that of a non-neurotensin patient, the neurotensin is judged.
7. The apparatus for differential diagnosis of a neurosyphilis according to claim 5 or 6, wherein the judgment criteria of the judgment module comprises: and if the SEMA7A content in the sample to be detected is obviously reduced compared with the non-neural syphilis patient, judging that the sample is neural syphilis.
8. The device for differential diagnosis of neurosyphilis according to claim 7, wherein the sample to be tested is blood, plasma or serum.
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