Use of MxA as a marker for diagnosing severe viral pneumonia-related diseases

Abstract

The present invention discloses the use of MxA or its binding antibody or fragment in the preparation of a diagnostic reagent for warning an individual of severe viral pneumonia. The diagnostic kit for diagnosing an individual as belonging to severe viral pneumonia contains a diagnostic reagent, the diagnostic reagent contains MxA or its binding antibody or fragment, and the MxA or its binding antibody or fragment is compared with the individual's blood, plasma or serum sample for detection. In the invention, the sensitivity and specificity of MxA binding antibody or its fragment in identifying severe viral pneumonia from the pneumonia population are 85% and 89.1%, and MxA is used as a biomarker for the diagnosis of severe viral pneumonia, and its diagnostic detection effect is very outstanding.

Description

Technical Field

[0001] The present invention belongs to the technical field of biological diagnostic products, and in particular to a use of MxA or its binding antibody or fragment in the preparation of a diagnostic reagent for early warning of severe viral pneumonia in an individual. Background Art

[0002] Viral pneumonia is one of the common diseases of community-acquired pneumonia in children, and it is also one of the most serious types. It mostly occurs in infants and young children aged 6 months to 5 years. Common respiratory viruses include respiratory syncytial virus, influenza virus, adenovirus, parainfluenza virus and rhinovirus. Emerging viruses include human metapneumovirus, bocavirus, new coronavirus, human avian influenza virus, etc. The clinical manifestations of severe viral pneumonia are repeated high fever, severe systemic poisoning symptoms, mental depression or irritability, shortness of breath, cyanosis of the lips and even respiratory failure. It has a long course of disease, is difficult to treat, and progresses rapidly. The mortality rate can be as high as 10%. Among the survivors, 14% to 60% may have sequelae of varying degrees, such as obliterative bronchiolitis (BO), which is currently one of the important causes of death and disability from pneumonia in infants and young children. To date, the pathogenesis of severe viral pneumonia is still unclear. It is believed that it may be related to the virus itself and the inflammatory response induced by the body. The activation of the immune system and the massive release of cytokines or inflammatory mediators play a vital role.

[0003] During viral infection, a cellular response to secrete type I interferons (IFNs) occurs. Type I IFNs participate in the innate immune response and have immunomodulatory, anti-cellular proliferation, and antiviral effects. Although sometimes used as a biological marker for viruses, type I IFNs have a short half-life in body fluids and have not shown good diagnostic efficacy. The antiviral activity of type I IFNs mostly comes from some proteins, mediated by the production of myxovirus resistance protein A (MxA). MxA was first described as an antiviral protein in influenza virus infection, and was subsequently found to have a broad spectrum of antiviral activity against a variety of RNA viruses and some DNA viruses.

[0004] To this end, we propose a method of using MxA or its binding antibody or fragment in the preparation of a diagnostic reagent for warning of severe viral pneumonia in individuals. Summary of the invention

[0005] The object of the present invention is to provide a method for preparing a diagnostic reagent for early warning of severe viral pneumonia in an individual by using MxA or its binding antibody or fragment, so as to solve the problems raised in the above background technology.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] A use of MxA or its binding antibody or fragment in the preparation of a diagnostic reagent for early warning of severe viral pneumonia in an individual, comprising the following:

[0008] Use of MxA or its binding antibody or fragment in the preparation of a diagnostic reagent for warning of severe viral pneumonia, wherein the diagnostic reagent warns whether an individual is at risk of developing severe viral pneumonia by detecting the MxA content in an individual sample.

[0009] Use of MxA or its binding antibody or fragment in the preparation of a diagnostic reagent for warning of severe viral pneumonia in an individual, wherein the diagnostic reagent diagnoses viral pneumonia by detecting the content of MXA in an individual sample and comparing it with a normal level.

[0010] Use of MxA or its binding antibody or fragment in the preparation of a diagnostic reagent for warning an individual of severe viral pneumonia, wherein the diagnostic reagent warns whether the individual is at risk of developing severe viral pneumonia by detecting the MxA content in an individual sample.

[0011] Use of MxA or its binding antibody or fragment in the preparation of a diagnostic reagent for warning of severe viral pneumonia in an individual, wherein the diagnostic reagent diagnoses viral pneumonia by detecting the content of MxA in an individual sample and comparing it with a normal level.

[0012] A diagnostic product for severe viral pneumonia-related diseases using MxA as a marker, including the following:

[0013] A diagnostic product for distinguishing whether an individual has non-severe viral pneumonia or severe viral pneumonia. The diagnostic product contains a diagnostic reagent, which includes an MxA-binding antibody or a binding antibody fragment. The binding antibody or the binding antibody fragment binds to MxA in an individual sample, and then detects the MxA content in the sample for diagnosis.

[0014] A diagnostic product for determining whether an individual suffers from viral pneumonia, the diagnostic product containing a diagnostic reagent, the diagnostic reagent containing an MxA binding antibody or a binding antibody fragment, and the binding antibody or the binding antibody fragment binds to MxA in an individual sample, thereby detecting the MxA content in the sample for diagnosis.

[0015] In a further embodiment, the diagnostic reagent can detect the content of MxA by any method based on its structure and / or component composition, including but not limited to electrophoresis, immunofluorescence, direct competition, indirect competition, ELISA, RIA, flow cytometry or immunochromatography.

[0016] The diagnostic scheme of MxA in severe viral pneumonia-related diseases includes the following:

[0017] A method for diagnosing whether an individual suffers from viral pneumonia, the method comprising the step of using a diagnostic reagent to detect the MxA content in an individual sample, and further comparing the detected MxA content with a normal level to further determine whether the individual suffers from viral pneumonia.

[0018] A method for diagnosing an individual to distinguish between non-severe viral pneumonia and severe viral pneumonia, the method comprising the step of using a diagnostic reagent to detect the MxA content in an individual sample to further distinguish between the individual to belong to non-severe viral pneumonia and severe viral pneumonia.

[0019] In a further embodiment, the diagnostic reagent can detect the content of MxA by any method based on its structure and / or component composition, including but not limited to electrophoresis, immunofluorescence, direct competition, indirect competition, ELISA, RIA, flow cytometry or immunochromatography.

[0020] In yet further embodiments, the sample from the individual is one or more of whole blood, plasma or serum taken from the individual.

[0021] In a further embodiment, the normal level can refer to the MxA level of a person who is not infected with the virus, such as the MxA level of a normal person. In the present invention, the normal level can be implemented based on whether the MxA level is greater than 20 ng / mL.

[0022] In a further embodiment, the diagnostic reagent is provided with a warning line identification mark for MxA, and the warning line identification mark is used for the user to compare whether the MxA content in the reference sample exceeds the normal level; the warning line identification mark preferably includes the test result corresponding to the sample MxA content of 18-25 ng / mL; the warning line identification mark most preferably includes the test result when the MxA content is 20 ng / mL.

[0023] In a further embodiment, the diagnostic reagent is provided with a warning line identification mark for MxA, and the warning line identification mark is used for the user to compare whether the MxA content in the reference sample exceeds the set level; the warning line identification mark preferably includes the test result corresponding to the sample MxA content of 90-110 ng / mL; the warning line identification mark most preferably includes the test result when the MxA content is 100 ng / mL.

[0024] Compared with the prior art, the beneficial effects of the present invention are: the sensitivity and specificity of MxA detection in distinguishing non-severe cases from the pneumonia population are 89.1% and 85%, and the sensitivity and specificity in distinguishing severe cases from the pneumonia population are 85% and 89.1%. MxA is used as a biomarker for the diagnosis of viral pneumonia, the identification of non-severe viral pneumonia and severe viral pneumonia, and its diagnostic detection effect is very outstanding. DETAILED DESCRIPTION

[0025] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0026] The technical concept of the present invention is to use MxA as a biomarker to develop diagnostic schemes, diagnostic products and uses for early warning of severe viral pneumonia.

[0027] Example 1, the present invention first provides the relevant use of MxA in the preparation of diagnostic reagents for early warning of severe viral pneumonia, including the following:

[0028] Use of MxA or its binding antibody or fragment in the preparation of a diagnostic reagent for warning of severe viral pneumonia, wherein the diagnostic reagent warns whether an individual is at risk of developing severe viral pneumonia by detecting the MxA content in an individual sample.

[0029] Use of MxA or its binding antibody or fragment in the preparation of a viral pneumonia diagnostic reagent, wherein the diagnostic reagent diagnoses viral pneumonia by detecting the content of MxA in an individual sample and comparing it with a normal level.

[0030] The use of MxA as a biomarker in a diagnostic reagent for early warning of severe viral pneumonia, wherein the diagnostic reagent warns whether an individual is at risk of developing severe viral pneumonia by detecting the MxA content in an individual sample.

[0031] Use of MxA as a biomarker in the preparation of a viral pneumonia diagnostic reagent, wherein the diagnostic reagent diagnoses viral pneumonia by detecting the content of MxA in an individual sample and comparing it with a normal level.

[0032] Example 2, the present invention further directly provides a diagnostic scheme for MxA in severe viral pneumonia-related diseases, including the following:

[0033] A method for diagnosing whether an individual suffers from viral pneumonia, the method comprising the step of using a diagnostic reagent to detect the MxA content in an individual sample, and further comparing the detected MxA content with a normal level to further determine whether the individual suffers from viral pneumonia.

[0034] A method for diagnosing an individual to distinguish between non-severe viral pneumonia and severe viral pneumonia, the method comprising the step of using a diagnostic reagent to detect the MxA content in an individual sample to further distinguish between the individual to belong to non-severe viral pneumonia and severe viral pneumonia.

[0035] Example 3: The third aspect of the present invention also provides a diagnostic product using MxA as a biomarker, including the following:

[0036] A diagnostic product for distinguishing whether an individual has non-severe viral pneumonia or severe viral pneumonia. The diagnostic product contains a diagnostic reagent, which includes an MxA-binding antibody or a binding antibody fragment. The binding antibody or the binding antibody fragment binds to MxA in an individual sample, and then detects the MxA content in the sample for diagnosis.

[0037] A diagnostic product for determining whether an individual suffers from viral pneumonia, the diagnostic product containing a diagnostic reagent, the diagnostic reagent containing an MxA binding antibody or a binding antibody fragment, and the binding antibody or the binding antibody fragment binds to MxA in an individual sample, thereby detecting the MxA content in the sample for diagnosis.

[0038] According to the specific implementation of the present invention and in combination with the knowledge in the art, in the uses, diagnostic methods or diagnostic products provided by the present invention above: the diagnostic reagent mentioned can detect the content of MxA by any method through its structure and / or component composition, including but not limited to electrophoresis, immunofluorescence, direct competition, indirect competition, ELISA, RIA, flow cytometry or immunochromatography.

[0039] According to the specific implementation of the present invention and in combination with the knowledge in the art, in the uses, diagnostic methods or diagnostic products provided by the present invention above: the individual sample mentioned is whole blood, plasma or serum taken from the individual.

[0040] According to the specific implementation of the present invention and in combination with the knowledge in the art, in the uses, diagnostic methods or diagnostic products provided by the present invention above: the normal level can refer to the MxA content level of people who do not have viral infection, such as the MxA content level of normal people. In the present invention, the normal level can be implemented based on whether the MxA content is greater than 20 ng / mL.

[0041] According to the specific implementation of the present invention and in combination with the knowledge in the art, the uses, diagnostic methods or diagnostic products provided by the present invention above: warning whether the individual is at risk of developing severe viral pneumonia, and whether the MxA content in the sample is greater than 100 ng / mL.

[0042] According to the specific implementation of the present invention and in combination with the knowledge in the art, in the uses, diagnostic methods or diagnostic products provided by the present invention above: the diagnostic reagent is provided with a warning line identification mark for MxA, and the warning line identification mark is used for the user to compare whether the MxA content in the reference sample exceeds the normal level; the warning line identification mark preferably includes the test result corresponding to the sample MxA content of 18-25 ng / mL; the warning line identification mark most preferably includes the test result when the MxA content is 20 ng / mL.

[0043] According to the specific implementation of the present invention and in combination with the knowledge in the art, in the uses, diagnostic methods or diagnostic products provided by the present invention above: to warn whether an individual is at risk of developing severe viral pneumonia, the diagnostic reagent is provided with a warning line identification mark for MxA, and the warning line identification mark is used for the user to compare whether the MxA content in the reference sample exceeds the set level; the warning line identification mark preferably includes the test result corresponding to the sample MxA content of 90-110 ng / mL; the warning line identification mark most preferably includes the test result when the MxA content is 100 ng / mL.

[0044] Clinical Validation

[0045] For specific clinical trial methods not listed, please refer to relevant national standards and common industry practices and will not be elaborated here.

[0046] 1. The experiment uses the MxA detection kit as the research and verification test object.

[0047] MxA Detection Kit The detection kit can be prepared by the method described in the Chinese invention patent with application number CN113866428A previously applied by the applicant of the present invention.

[0048] 2. The test participants were selected from 57 cases of viral pneumonia and 10 cases of surgical indirect hernia surgery control cases from the Children's Hospital Affiliated to Chongqing Medical University for validation research. The diagnostic criteria for viral pneumonia were in accordance with Zhu Futang's Practical Pediatrics, and the diagnostic criteria for severe viral pneumonia were in accordance with the Guidelines for the Management of Community-Acquired Pneumonia in Children (Revised in 2013) formulated by the Respiratory Group of the Pediatric Branch of the Chinese Medical Association.

[0049] 3. Test methods:

[0050] 1. The MxA test kit was used to test 67 patients: the MxA content of 20ng / mL was used as the diagnostic threshold for viral pneumonia. The MxA content of 100ng / mL was used as the diagnostic threshold for severe viral pneumonia. If the test results show that the MxA content in the individual sample is greater than 20ng / mL and not greater than 100ng / mL, it is marked as viral pneumonia. If the test results show that the MxA content in the individual sample is greater than 100ng / mL, it is marked as severe viral pneumonia. If the MxA content is not greater than 20ng / mL, it is marked as normal.

[0051] 2. Use conventional means to diagnose the conditions of 57 patients: Use a variety of conventional clinical methods to diagnose the conditions of 57 patients and determine their true conditions.

[0052] 3. Compare and contrast the diagnostic marker results of the MxA detection kit with those of conventional methods, and statistically analyze the detection sensitivity and specificity.

[0053] 4. Test results:

[0054] According to the above test method, the test verification results are shown in Table 1 and Table 2 below:

[0055] Table 1 Results of 67 cases in Children's Hospital Affiliated to Chongqing Medical University

[0056] Case series MxA ng / mL Admission diagnosis Classification 1 4.85 Indirect hernia No virus infection group 2 1.37 Indirect hernia No virus infection group 3 7.13 Indirect hernia No virus infection group 4 3.66 Indirect hernia No virus infection group 5 8.76 Indirect hernia No virus infection group 6 3.49 Indirect hernia No virus infection group 7 5.37 Indirect hernia No virus infection group 8 2.36 Indirect hernia No virus infection group 9 4.53 Indirect hernia No virus infection group 10 9.82 Indirect hernia No virus infection group 11 20.45 Viral pneumonia Non-severe group 12 39.95 Viral pneumonia Non-severe group 13 36.88 Viral pneumonia Non-severe group 14 36.33 Viral pneumonia Non-severe group 15 44.25 Viral pneumonia Non-severe group 16 42.78 Viral pneumonia Non-severe group 17 78.23 Viral pneumonia Non-severe group 18 69.54 Viral pneumonia Non-severe group 19 38.36 Viral pneumonia Non-severe group 20 145.23 Viral pneumonia Non-severe group 21 112.23 Viral pneumonia Non-severe group 22 79.65 Viral pneumonia Non-severe group 23 156.33 Viral pneumonia Non-severe group 24 80.42 Viral pneumonia Non-severe group 25 68.98 Viral pneumonia Non-severe group 26 65.25 Viral pneumonia Non-severe group 27 64.54 Viral pneumonia Non-severe group 28 94.23 Viral pneumonia Non-severe group 29 49.31 Viral pneumonia Non-severe group 30 76.32 Viral pneumonia Non-severe group 31 30.34 Viral pneumonia Non-severe group 32 55.56 Viral pneumonia Non-severe group 33 75.32 Viral pneumonia Non-severe group 34 24.45 Viral pneumonia Non-severe group 35 21.29 Viral pneumonia Non-severe group 36 57.76 Viral pneumonia Non-severe group 37 49.26 Viral pneumonia Non-severe group 38 25.62 Viral pneumonia Non-severe group 39 46.62 Viral pneumonia Non-severe group 40 115.32 Viral pneumonia Non-severe group 41 36.89 Viral pneumonia Non-severe group 42 79.72 Viral pneumonia Non-severe group 43 67.88 Viral pneumonia Non-severe group 44 45.23 Viral pneumonia Non-severe group 45 42.44 Viral pneumonia Non-severe group 46 76.52 Viral pneumonia Non-severe group 47 73.46 Viral pneumonia Non-severe group 48 110.22 Viral pneumonia Severe group 49 208.11 Viral pneumonia Severe group 50 187.04 Viral pneumonia Severe group 51 207.21 Viral pneumonia Severe group 52 133.04 Viral pneumonia Severe group 53 389.23 Viral pneumonia Severe group 54 456.21 Viral pneumonia Severe group 55 184.23 Viral pneumonia Severe group 56 146.23 Viral pneumonia Severe group 57 94.7 Viral pneumonia Severe group 58 78.6 Viral pneumonia Severe group 59 338.36 Viral pneumonia Severe group 60 573.82 Viral pneumonia Severe group 61 150.02 Viral pneumonia Severe group 62 87.32 Viral pneumonia Severe group 63 145.23 Viral pneumonia Severe group 64 212.23 Viral pneumonia Severe group 65 179.65 Viral pneumonia Severe group 66 356.33 Viral pneumonia Severe group 67 187.58 Viral pneumonia Severe group

[0057] 5. Results and Statistics

[0058] The specificity and sensitivity in each case are statistically analyzed in Table 1.

[0059] Statistical Table 1 Data:

[0060] ① The sensitivity and specificity of MxA detection in diagnosing viral pneumonia infection in all populations, and the test results when the warning value MxA content is 20ng / mL

[0061] Sensitivity: 57 cases of systemic infection were diagnosed routinely upon admission, of which 57 were correctly detected by MxA test, with a sensitivity of 100%;

[0062] Specificity: 10 cases of non-viral infection were diagnosed routinely on admission, of which 10 were correctly detected by MxA test, with a sensitivity of 100%;

[0063] ② The sensitivity and specificity of MxA detection in distinguishing non-severe viral pneumonia from severe viral pneumonia. The detection result when the warning value of MxA content is 100 ng / mL 1)

[0065] Thirty-seven cases of non-severe viral pneumonia were routinely diagnosed upon admission, of which 33 were correctly detected by MxA testing;

[0066] Twenty cases of severe viral pneumonia were routinely diagnosed upon admission, of which 17 were correct by MxA testing;

[0067] That is, the sensitivity and specificity of MxA detection in distinguishing non-severe cases from the pneumonia population were 89.1% and 85%, and the sensitivity and specificity of MxA detection in distinguishing severe cases from the pneumonia population were 85% and 89.1%.

[0068] It can be seen that the diagnostic detection effect of MxA as a biomarker for the diagnosis of viral pneumonia, non-severe viral pneumonia and severe viral pneumonia is very outstanding.

[0069] In addition, for the convenience of clinical use, warning line identification marks for MxA viral pneumonia, or non-severe viral pneumonia and severe viral pneumonia can be further designed on the MxA detection kit. The warning line identification marks can, for example, correspond to the test result pattern when the sample MxA content is 20 ng / mL and the test result pattern when the sample MxA content is 100 ng / mL.

[0070] It will be apparent to those skilled in the art that the invention is not limited to the details of the exemplary embodiments described above and that the invention can be implemented in other specific forms without departing from the spirit or essential features of the invention. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations within the meaning and scope of the equivalent elements of the claims be included in the invention. Any marking in a claim should not be considered as limiting the claim to which it relates.

[0071] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. Use of an antibody that binds to MxA in the preparation of a diagnostic reagent for warning an individual of severe viral pneumonia, comprising measuring the MxA biomarker in a sample of blood, plasma or serum taken from the individual, wherein the reagent for detecting MxA is an antibody that binds to MxA, the individual being suspected of being at risk of severe viral pneumonia, wherein determining whether the individual is at risk for developing severe viral pneumonia comprises determining whether the concentration of MxA in the sample is greater than 100 ng / mL; The normal level is based on an MxA content of 20 ng / mL to diagnose whether an individual has viral pneumonia.

2. Use of the MxA binding antibody according to claim 1 in the preparation of a diagnostic reagent for early warning of severe viral pneumonia in an individual, characterized in that: The content of MxA was detected by electrophoresis, immunofluorescence, ELISA, RIA, FCM or immunochromatography.

3. Use of the MxA binding antibody according to claim 1 in the preparation of a diagnostic reagent for early warning of severe viral pneumonia in an individual, characterized in that: The diagnostic reagent is provided with a warning line identification mark of MxA, and the warning line identification mark is used for the user to compare whether the MxA content in the reference sample exceeds the normal human level.