JP2013205242A - Method for examining prognosis of brain infarction by il-8 protein/nihss - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method with which a prognosis of brain infarction can be examined simply and efficiently.SOLUTION: An examination method for predicting a prognosis of brain infarction in a test animal includes a step for measuring a protein content of IL-8 protein in a blood specimen which is sampled from the animal. The examination method for predicting the prognosis of brain infarction in the test animal also includes a step for measuring the protein content of IL-8 protein and NIHSS in the blood specimen which is sampled from the animal.

Description

  The present invention relates to a prognosis test method for cerebral infarction and a prognosis test kit for cerebral infarction.

  Stroke is a general term for cerebrovascular disorders such as cerebral infarction, cerebral hemorrhage, and subarachnoid hemorrhage. Of these stroke types, cerebral infarction has been relatively increasing recently. Thrombolytic therapy, anticoagulant therapy, antiplatelet therapy, administration of cerebral protective drugs, etc. are used as drug therapy in the acute phase of cerebral infarction. Surgical surgery is performed for patients with severe or hemorrhagic cerebral infarction. Is done. Cerebral infarction symptoms are strongest in the acute phase and then gradually improve. This is because the necrotic brain tissue is swollen and the surrounding brain tissue is also compressed and damaged. As the swelling subsides, the surrounding tissues recover their function and the symptoms are fixed. However, since free radicals released from cerebral ischemic sites have the function of necrosing surrounding tissues, continuous treatment is required to improve the functional prognosis even after the acute phase.

  The clinical types of cerebral infarction include lacunar infarction that develops when the intracerebral small artery is occluded, atherothrombotic cerebral infarction that develops when the intracerebral aorta is occluded by atheroma, and a thrombus in the heart becomes an obturator. Examples include cardiogenic cerebral embolism that develops by blocking an artery. Since appropriate treatment methods for the acute and chronic phases differ depending on these clinical types, a method for rapidly diagnosing the clinical type of patients with cerebral infarction has been demanded.

As a method of classifying clinical types of cerebral infarction, clinical symptom observation and echocardiography, MRI, MRA, cervical vascular echo, etc. are performed, and a flowchart of clinical diagnosis of cerebral infarction based on TOAST classification (Non-patent Document 1), etc. As a result of simultaneous measurement of molecular markers such as CRP, D-Dimer, RAGE, MMP-9, S100B, BNP within 24 hours of cerebral infarction, Although it has been disclosed that cardiogenic cerebral embolism can be classified as another disease type by setting a specific cutoff value with BNP and D-Dimer (Non-patent Document 2), etc. There was a need for molecular markers that could be diagnosed.

The prognosis of cerebral infarction is to evaluate the degree of movement disorder in daily life by the modified Rankin Scale (Non-patent Document 3) or Barthel Index (Non-patent Document 4) between 3 months or 1 year after onset. It is judged by. Predicting the prognosis in advance is important for early determination of a treatment method for cerebral infarction. Since the prognosis depends greatly on the severity of the acute phase, NIH Stroke Scale (NIHSS) (Non-Patent Document 6), which is an evaluation index of severity, is helpful.
This alone was not enough and was just a reference. Moreover, although it is disclosed that death can be predicted by determining a specific cutoff value with BNP in plasma measured at the time of admission (Non-patent Document 5), a molecular marker for predicting the degree of disability when surviving is disclosed. It was sought after.

Lee, L.J. et al., Stroke, 1081-1089 (2000) Montaner et al., Stroke, 2280-2287 (2008) van Swieten, J.C et al., Stroke, 604-607 (1988) Mahoney, F.L et al., Maryland State Med J, 61-65 (1965) Shibazaki, K et al., Inter Med, 1601-1604 (2009) Lyden, P.D. et al., Stroke 1310-1317 (2001)

  In view of the above problems, an object of the present invention is to provide a method for quickly and surely prognosing a cerebral infarction using serum, plasma, or the like of a cerebral infarction patient or the like.

As a result of intensive studies to solve the above-mentioned problems, the present inventors have obtained the knowledge that the amount of IL-8 protein in the patient plasma differs in patients with different prognosis of cerebral infarction, and further, NIHSS which is an index of severity As a result, the inventors have found that the accuracy of prognosis is improved when combined with the above, and found that the amount of IL-8 protein and NIHSS are useful indicators for the examination of cerebral infarction, thereby completing the present invention.

That is, the gist of the present invention is as follows.
(1) A test method for predicting the prognosis of cerebral infarction in an animal using IL-8 protein amount as an index, comprising the step of measuring the protein amount of IL-8 protein in a blood sample collected from the subject animal .
(2) The blood sample collected from the test animal is collected from the test animal at any time between the 2nd day and the 20th day after the onset of cerebral infarction-like symptoms. The testing method for predicting the prognosis of cerebral infarction according to (1).
(3) For predicting the prognosis of cerebral infarction in an animal using IL-8 protein mass and NIHSS as indices, including a step of measuring IL-8 protein mass and NIHSS in a blood sample collected from the test animal Inspection method.
(4) The blood sample collected from the test animal is collected from the test animal at any time between the 2nd day and the 20th day after the onset of cerebral infarction-like symptoms. The examination method for predicting the prognosis of cerebral infarction characterized by (2).
(5) A kit for examination of cerebral infarction comprising a reagent capable of measuring the protein amount of IL-8 protein.

According to the present invention, IL-8 protein is provided as a molecular marker for cerebral infarction examination by utilizing the fact that the concentration of IL-8 protein in the plasma of cerebral infarction patients differs between cerebral infarction patients with different prognosis. The By using the IL-8 protein marker alone or in combination with NIHSS, which is an indicator of severity, it is possible to predict cerebral infarction quickly and reliably.

Hereinafter, embodiments of the present invention will be described in detail.

(1) Cerebral infarction examination method using IL-8 protein marker A first subject of the present invention is a subject (animal) comprising a step of measuring the amount of IL-8 protein in a blood sample collected from the subject animal. This is a test method for predicting the prognosis of cerebral infarction.
In the present specification, “cerebral infarction” is a disease in which the cerebral artery is blocked by a thrombus or embolus and becomes ischemic, and the brain tissue is necrotized and damaged, and a lacunar infarction that develops due to blocking of a small intracerebral artery. These include both atherothrombotic cerebral infarction that develops when the intracerebral aorta is occluded by atheroma, and cardiogenic cerebral embolism that develops when the intracerebral artery is occluded to occlude the intracerebral artery. In addition, cerebral infarction includes any of acute phase, subacute phase, recovery phase (chronic phase) and the like.

  In the present specification, the “IL-8 protein” means an IL-8 (Interleukin 8) protein that is a target for measuring the content in a blood sample in the test method of the present invention, and is a protein derived from a test animal. However, specific examples of the human-derived protein include proteins represented by the specific amino acid sequences described below. Furthermore, fragments, derivatives, and mutants of proteins having the same functions as these are also included.

  In the above examination method, the “test animal” may be any animal that develops cerebral infarction, and specifically includes rodents such as humans, monkeys, and rats. The test animal may have any cerebral infarction, and the test method for cerebral infarction in that case is particularly preferably performed in humans suspected of having cerebral infarction or after cerebral infarction onset to 20 days later. Is called.

  In the test method of the present invention, the time for collecting a blood sample from the test animal is particularly preferably 2 to 20 days after the onset of cerebral infarction, and particularly 3 to 15 days after the onset of cerebral infarction.

  The “blood sample” collected from the test animal is not particularly limited as long as it contains IL-8 protein and can measure its concentration. Specifically, EDTA plasma, citrate Any of plasma such as plasma, serum, and whole blood may be used, but among these, EDTA plasma can be easily collected, is easily stored, and is preferably used because of its large collection amount. The timing for collecting the blood sample from the test animal may be any timing as long as the prognosis for cerebral infarction is diagnosed. Since cerebral infarction changes in symptoms and pathology from onset to onset, it will be described in detail below.

  In the test method of the present invention, the IL-8 protein that is a target for measuring the content in a blood sample collected from a test animal is, for example, a database of the National Center for Biotechnology Information (NCBI) in the case of humans. Etc. having the amino acid sequence registered under NP_000575.1. The amino acid sequence is human, but when the test animal is different, a homologous protein derived from the animal becomes the protein to be measured.

The method for measuring the content of the protein in the blood sample is not particularly limited as long as the content of the protein can be measured. For example, an existing immunoassay method using an antibody of the protein or a chromatographic method. All components captured on a chromatographic carrier (eg, cation exchanger, anion exchanger, hydrophobic chromatographic carrier, metal ion, etc.) under certain conditions using a combination of holographic techniques and time-of-flight mass spectrometry (TOF-MS) For example, a method of measuring the mass of the solution at once, a two-dimensional gel electrophoresis method, or the like is used. As the immunoassay, a method of quantitative detection according to an enzyme immunoassay, a method of measurement by a fluorescence immunoassay, a chemiluminescence immunoassay, or the like is preferable. The enzyme immunoassay method is a detection method using an enzyme as a labeling substance among the labeled immunoassay methods. In addition, selecting an enzyme-linked immunosorbent assay (ELISA) method using an immunosolvent
Particularly preferred. Among the ELISA methods, the sandwich method is mentioned as one of the particularly preferable measurement modes in consideration of the convenience of operation, economic convenience, and general versatility in clinical examination. These measurement methods include, for example, the New Chemistry Laboratory (Edited by the Japanese Biochemical Society; Tokyo Kagaku Dojin), Molecular Cloning, A Laboratory Manual (T. Maniatis et al., Cold Spring Harbor Laboratory (2001)), Antibodies-A Laboratory. Manual (E. Harlow, et al., Cold
It can be carried out according to the method described in a general experiment such as Spring Harbor Laboratory (1988) or the like.

Antibodies and the like (including antibody fragments) used in the above measurement can be obtained by a known method using the target IL-8 protein as an antigen. However, it is not necessary to be produced using the target IL-8 protein as an antigen, and any protein may be used as long as it exhibits at least cross-reactivity with the protein and can measure its content. As such an immunoassay method, for example, there is a method in which measurement using the Human Map ^™ v.1.6 method is performed by requesting Rules Based or the like.

As a blood sample used in the method of the present invention, a blood sample immediately after being collected from a test animal is preferably used for the measurement, but a stored sample may be used. The method for storing the blood sample is not particularly limited as long as the amount of molecular marker for cerebral infarction examination does not change. For example, low temperature conditions such as 0 to 10 ° C. that do not freeze, dark conditions, and no vibration conditions. preferable. In the case of unavoidably freezing, a method capable of avoiding marker molecule decomposition, oxidation reaction, etc., such as deep freezing, is preferable.

  In the test method of the present invention, the content of the IL-8 protein marker in a blood sample collected from a test animal (in this specification, this may be referred to as “specimen”) is measured. Using this as an index, the prognosis of cerebral infarction can be examined. In addition, it is possible to test more accurately by using a composite index that correlates observations of clinical symptoms and results such as echocardiography, MRI, MRA, and cervical vascular echo.

(1-1) Test method for predicting prognosis of cerebral infarction using IL-8 protein marker Test method for predicting prognosis of cerebral infarction of subject (animal) using IL-8 protein marker of the present invention A specific method will be described. The prognosis of cerebral infarction is indicated by the degree of injury due to cerebral infarction of the patient after a certain period of time after the onset of cerebral infarction. The degree of failure can be assessed using an already established indicator known per se. Indicators include, for example, Japan's modified Rankin Scale (mRS) criteria (Yukito Shinohara et al., MRS Reliability Study Group, Study on Reliability of Modified Rankin Scale-Introduction of Japanese Version Criteria and Questionnaire, Stroke 2007
; 29: 6-13) is used.

After a certain period of time, it may be any time after onset, but from the viewpoint of assessing the state of rehabilitation of a subject (animal), for example, it is generally evaluated from 3 months to 1 year after the onset Is. In the prediction of prognosis, taking into consideration the usefulness of the prediction method, the amount of IL-8 protein in the blood sample obtained from the subject 2 to 20 days after the onset, and the test 3 months after the onset A method for predicting a person's condition is preferable.

In the measurement method of the present invention, the amount of IL-8 protein in a blood sample at the time of measurement for each patient who has been clinically determined in advance at the above-mentioned mRS at the time of the prediction purpose is measured, and the mRS to be distinguished at the time of the prediction purpose. In the meantime, the cutoff value which distinguishes both mRS groups by the above-mentioned method is set. More specifically, for example, in a case where a patient with mRS of 0 to 1 after 3 months of onset is distinguished from a patient with 2 to 5 of mRS, a patient with mRS of 0 to 5 3 months after onset (during prediction purposes) 2-8 days after the onset of the disease, preferably 3-15 days later (for measurement purposes), the amount of IL-8 protein in the blood sample is measured. Set a cut-off value between the patient's measurements. Thereafter, a sample at the time of measurement is collected, the content of the IL-8 protein marker in the sample is measured, and the absolute value is compared with the cut-off value set by the above method. The state of the subject (animal) at the time of purpose can be predicted.

  The cut-off value refers to a value determined when a target disease group and a non-disease group are determined by paying attention to a certain substance. When determining the target disease and non-disease, it is negative if it is below the cut-off value, positive if it is above the cut-off value, or positive if it is below the cut-off value, and if it is above the cut-off value The disease can be determined as negative (Kanai Masamitsu, Clinical Laboratory Proposal Kanehara Publishing Co., Ltd.).

  The index used for the purpose of evaluating the clinical usefulness of the cutoff value includes sensitivity and specificity. A certain population is determined using a cut-off value, and among the diseased patients, a (true positive) is positive in the determination, and b (false negative) is negative in the determination while being a disease patient. A / (a + b) when c (false positive) indicates that the patient is not a disease patient but is positive in the determination and d (true negative) indicates that the patient is not a disease patient and is negative in the determination. Can be represented as sensitivity (true positive rate) and d / (c + d) as specificity (true negative rate).

The distribution of measured values of IL-8 levels in blood samples between the target mRS and other mRS specimens usually has some overlap. Therefore, sensitivity and specificity change by raising or lowering the cutoff value. Decreasing the cut-off value increases the sensitivity, but the specificity decreases, and increasing the cut-off value decreases the sensitivity, but increases the specificity. As a determination method, it is preferable that both sensitivity and specificity are higher. A determination method in which the values of sensitivity and specificity do not exceed 0.5 is not recognized as useful.

  The method for setting the cut-off value is to set 95% of the distribution of the non-disease group as one of the values at both ends from the center as the cut-off value. When the distribution of the non-disease group shows a normal distribution And a method of setting an average value + 2 times standard deviation (SD) or an average value−2SD as a cut-off value.

  In general, in order to determine whether or not a diagnostic method is useful, the sensitivity and specificity vary depending on the cutoff value set as described above, so the sensitivity and specificity at a certain cutoff value are simply used. Rather than evaluating, it is desirable to evaluate with an index that maintains high sensitivity and specificity when the cut-off value is raised or lowered, for example, the AUC value of the ROC curve. The AUC value of the ROC curve becomes 1 when a cutoff value exists such that both sensitivity and specificity are 100%, and approaches 0.5 when the diagnostic performance is not good. Therefore, when the performance of a certain diagnostic method is judged by the AUC value of the ROC curve, it can be evaluated that the method is suitable as a diagnostic method if it is 0.7 or more. Since the IL-8 protein marker described in the present invention also has an ROC curve AUC value of 0.7 or more, it is used in the cerebral infarction examination method described later.

In the prediction method of the present invention, particularly preferably 2 to 20 days after the onset, more preferably 3 months after the onset by measuring the amount of IL-8 protein in a sample collected from a subject on the 3rd to 15th day of onset Examples include a method for predicting whether the mRS of the subject in the vicinity is 2 to 5 or less than 1.

  Thus, a subject (animal) diagnosed with cerebral infarction by the method of the present invention has a very good prognostic course and therapeutic effect by receiving treatment suitable for each disease.

(1-2) Test method for predicting prognosis of cerebral infarction using IL-8 protein marker and NIHSS The second aspect of the present invention is the amount and severity of IL-8 protein in a blood sample collected from a test animal. It is an inspection method for predicting the prognosis of cerebral infarction in a subject (animal), including a step of measuring NIHSS as an index. By using not only the IL-8 protein marker but also NIHSS in combination, the prognosis prediction of the test animal can be diagnosed more accurately. In this method, the test animal and the blood sample obtained from the animal are the same as in the above-described test method using IL-8. In the test method of the present invention, the time for collecting a blood sample from the test animal is particularly preferably 2 to 20 days after the onset of cerebral infarction, and particularly 3 to 15 days after the onset of cerebral infarction.

In the test method of the present invention, the IL-8 protein, which is a target for measuring the content in a blood sample collected from a test animal, is as described above. NIHSS is an index that is calculated by adding the degree of clinical symptoms of each defined item, and is used to evaluate the severity of stroke patients. The measured IL-8 protein mass and NIHSS and the prognosis of cerebral infarction can be correlated according to a known statistical model such as logistic regression, k-nearest neighbor method, linear multiple regression analysis or the like.
The method for measuring the content of IL-8 protein in the blood sample of the protein is as described above.

In the prediction method of the present invention, it is particularly preferable to measure IL-8 protein amount and NIHSS at the same time in a sample collected from a subject 2 to 20 days after onset, more preferably 3 to 15 days after onset. And a method for predicting whether the mRS of the subject in the vicinity of 3 months after the onset is 2 to 5 or 1 or less. In addition, a method of specifically predicting the mRS of the subject in the vicinity of 3 months after the onset by measuring the IL-8 protein amount in the sample and NIHSS at the same time is also included.
Thus, a subject (animal) diagnosed with cerebral infarction by the method of the present invention has a very good prognostic course and therapeutic effect by receiving treatment suitable for each disease.

(2) Kit for measuring cerebral infarction The present invention further includes a kit for predicting the prognosis of the cerebral infarction. The contents of the kit are configured by a combination of equipment or reagents. However, the configuration or form of the kit is not limited as long as it includes a substance that is essentially the same as each component described below, or a substance that is essentially the same as a part thereof. Even if they are different, they are included in the kit of the present invention. As a reagent, for example, when measuring an IL-8 protein marker by an immunoassay, an anti-IL-8 protein marker antibody is included. In addition, a biological sample diluent, an antibody-immobilized solid phase, a reaction buffer, a washing solution, a labeled secondary antibody or an antibody fragment thereof, a labeled detection reagent, a standard substance, and the like are also included as necessary. Examples of the diluted solution of the biological sample include an aqueous solution containing a protein such as BSA or casein in a surfactant or a buffer.

  As the antibody-immobilized solid phase, a material in which various polymer materials are shaped to suit the application and anti-molecular marker antibodies or antibody fragments thereof are solid-phased is used. Shapes are tubes, beads, plates, fine particles such as latex, sticks, etc., and materials are polystyrene, polycarbonate, polyvinyl toluene, polypropylene, polyethylene, polyvinyl chloride, nylon, polymethacrylate, gelatin, agarose, cellulose, polyethylene terephthalate, etc. Polymer materials, glass, ceramics and metals. Examples of the method for immobilizing an antibody include known methods such as a physical method and a chemical method, or a combination method thereof. For example, a 96-well polystyrene immunoassay microterplate prepared by subjecting an antibody or antibody fragment to a hydrophobic solid phase can be used.

  Any reaction buffer may be used as long as it provides a solvent environment for the binding reaction between the antibody on the antibody-immobilized solid phase and the antigen in the biological sample. However, surfactants, buffers, BSA, Reaction buffers containing proteins such as casein, preservatives, stabilizers, reaction accelerators and the like can be mentioned.

  As the labeled secondary antibody or antibody fragment thereof, the antibody or antibody fragment used in the present invention is labeled with a labeling enzyme such as horseradish peroxidase (HRP), bovine small intestine alkaline phosphatase, β-galactosidase, or a buffering agent. , BSA, casein and other proteins, preservatives and the like are used.

  As a reagent for detecting a labeled substance, for example, horseradish peroxidase according to the labeling enzyme, a substrate for absorption measurement such as tetramethylbenzidine or orthophenylenediamine, or a fluorescence such as hydroxyphenylpropionic acid or hydroxyphenylacetic acid. If the luminescent substrate such as a substrate or luminol is alkaline phosphatase, an absorbance measurement substrate such as 4-nitrophenyl phosphate, a fluorescent substrate such as 4-methylumbelliferyl phosphate, and the like can be given.

  EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples, but the present invention is not limited to these examples.

[Example 1] Evaluation of test method for predicting the prognosis of cerebral infarction using IL-8 protein marker The prognosis of cerebral infarction is a disorder caused by cerebral infarction of the patient after a certain period of time after the onset of cerebral infarction. It is indicated by the degree. Japanese version of the modified Rankin Scale (3 months after the onset of cerebral infarction)
mRS) Criteria (Yukito Shinohara et al. mRS Reliability Study Group, Study on Reliability of Modified Rankin Scale-Introduction of Japanese Version Criteria and Questionnaire, Stroke 2007; 29: 6-13) 0-5 For patients with a good prognosis, 79 patients with good prognosis (mRS 0-1) and 68 patients with poor prognosis (mRS 2 or more) 3 days after the onset of cerebral infarction (indicated as “3 days” in the table), 7 Blood was collected one day later (described as “7 days” in the table) and 14 days later (described as “14 days” in the table) to obtain EDTA plasma.
The concentration of IL-8 protein in EDTA plasma was measured by an immunoassay performed by requesting Rules Based Co., Ltd. for measurement using the Human Map ^™ v.1.6 method. Specifically, a standard curve was prepared using the standard protein attached to the kit, and the concentration of IL-8 protein in plasma was calculated from this standard curve.
As a control, it is already known that the peak concentration at 1 week after onset is associated with prognosis (Smith CJ, et al. BMC Neurol 2004; 4: 2. [PubMed: 14725719]) The CRP protein was measured in the same manner.
Next, Fawcett, T. (2004) ROC Graphs:
The AUC value of the ROC curve was calculated according to the method described in Notes and Practical Considerations for Researchers. The results are shown in Table 1.
It is shown that a value exceeding 0.7 at each time of blood collection is useful as a test method for determining the prognosis of a cerebral infarction patient. In other words, on the 3rd, 7th, and 14th days of the onset of cerebral infarction, by measuring the concentration of IL-8 protein in EDTA plasma, the Japanese modified Rankin Scale (mRS) criteria of 0 and It turned out that the patient who becomes 1 and 2 or more can be discriminated.

[Example 2] Evaluation of test method for predicting prognosis of cerebral infarction using IL-8 protein marker and NIHSS (1)
As in Example 1, 79 patients with good prognosis (mRS 0-1) and 68 patients with poor prognosis (mRS 2 or more) 3 days after onset of cerebral infarction (described as “3 days” in the table), 7 days after (It is described as “7 days” in the table) and 14 days later (described as “14 days” in the table) to obtain EDTA plasma, and in the same manner as in Example 1, EDTA IL-8 protein in plasma and IL-6 shown in Example 1
And the concentration of CRP protein was measured and calculated. NIHSS was also measured at the same time as blood collection.
Next, in order to evaluate the discrimination performance at each blood sampling point, NIHSS, IL-8, Il-6, and CRP proteins and any one or two variables of blood parameters were used as explanatory variables, and prognosis was used as a target variable. A prediction was made by regression. NIHSS performs a square root transformation to include zero,
Logarithmic transformation was performed for other protein amounts. In order to adjust the degree of overlearning according to the number of markers, ROC analysis was performed with the predicted value of 5-fold cross-validation, and the AUC value was calculated. The results are shown in Table 2.
It is shown that an AUC value exceeding 0.7 at each blood sampling time is useful as a test method for determining the prognosis of a cerebral infarction patient. However, all of the above proteins are already known to be effective alone as protein markers for predicting the prognosis of cerebral infarction. Is effective as a prognostic marker combination of cerebral infarction, so that the concentration of IL-8 protein and NIHSS in EDTA plasma were measured at about 3 days, 7 days, and 14 days from the onset of cerebral infarction.
The accuracy of the Japanese modified Rankin Scale (mRS) criteria 0 and 1 after 2 months, and the discrimination of patients who become 2 or more, is improved compared to the case where it is performed alone, and it turns out that it is an effective measurement method. It was.

[Example 3] Evaluation of test method for predicting the prognosis of cerebral infarction using IL-8 protein marker and NIHSS (2)
In Example 2, the variables were transformed and logistic regression was used for the statistical model. However, the statistical model was converted to the k-nearest neighbor method (Belur V. Dasarathy, editor (1991) Nearest Neighbor (NN) Norms: Except for NN Pattern Classification Techniques), the relationship between the amounts of NIHSS and IL-8 protein markers and the prediction of cerebral infarction was analyzed in the same manner as in Example 2. The results are shown in Table 3.
As is clear from Table 3, even when the statistical model is the k-nearest neighbor method, the IL-8 protein concentration and NIHSS in EDTA plasma were measured on the 3rd, 7th, and 14th days of the onset of cerebral infarction. About 3
It is shown that it is an effective measurement method with improved accuracy compared with the case where patients who become 0, 1 and 2 or more of Japanese version Modified Rankin Scale (mRS) criteria after a month are compared. It was.

[Example 4] Evaluation of test method for predicting prognosis of cerebral infarction using IL-8 protein marker and NIHSS (3)
In Example 2, variables were transformed and logistic regression was used for the statistical model. However, NIHSS and NIHSS were the same as Example 2 except that the same logistic regression as Example 2 was performed without performing variable transformation. The relationship between the amount of IL-8 protein marker and the prediction of cerebral infarction was analyzed. The results are shown in Table 3.
As is clear from Table 3, IL-8 protein concentration and NIHSS in EDTA plasma were measured at 3 days, 7 days, and 14 days after the onset of cerebral infarction even after logistic regression without variable exchange. By doing this, the Japanese modified Rankin Scale (mRS) criteria 0 and 1 after about 3 months
It was shown that the accuracy of the determination of the patient who becomes 2 or more is improved compared with the case where the determination is performed alone, and this is an effective measurement method.

[Example 5] Evaluation of test method for predicting prognosis of cerebral infarction using IL-8 protein marker and NIHSS (4)
Japanese modified Rankin Scale (mRS) criteria (Yukito Shinohara et al. MRS Reliability Study Group, Study on Reliability of modified Rankin Scale-Introduction of Japanese Version Criterion and Questionnaire, Stroke 2007; 29: 6-13) Blood samples were collected from 0 to 5 patients using mRI, that is, mRS0 31 people, mRS1 48 people, mRS2 29 people, mRS3 17 people, mRS4 21 people and mRS51 people 1 day after the onset of cerebral infarction. Plasma was obtained. Subsequently, in the same manner as in Example 1, the concentrations of IL-8 protein, CRP protein, and IL-6 protein in NIHSS and EDTA plasma were measured.
Linear multiple regression analysis with 1 to 2 variables of IL-8 protein mass, NIHSS, CRP protein mass, and IL-6 protein mass as explanatory variables and mRS as objective variable (Annette J. Dobson “Introduction to Generalized Linear Model”) Publishing, 2008). Variable conversion was performed in the same manner as in Example 1, and mRS was subjected to square root conversion. Next, the value of Spearman's rank correlation between the predicted value of the linear multiple regression analysis and the square root transformed mRS was calculated.
The results are shown in Table 4. As is apparent from Table 4, it was shown that when NIHSS was used alone, mRS indicating the prognosis of the patient about 3 months after onset could be predicted. In addition, when NIHSS and CRP protein, NIHSS and IL-6 protein, and NIHSS and IL-8 protein marker were combined, the prediction accuracy was improved compared to when they were used alone. It has been shown to be effective in predicting mRS indicating the patient's prognosis after about a month.

  According to the method of the present invention, it is possible to perform a prognostic test for cerebral infarction, which is useful in the medical and diagnostic fields.

Claims (5)

  A test method for predicting the prognosis of cerebral infarction in an animal using IL-8 protein mass as an index, comprising measuring the protein mass of IL-8 protein in a blood sample collected from a test animal.   The blood sample collected from the test animal is collected from the test animal at any time between the second day after the onset of cerebral infarction-like symptoms and the 20th day after the onset. The test method for predicting the prognosis of cerebral infarction according to claim 1.   A test method for predicting the prognosis of cerebral infarction in an animal using IL-8 protein mass and NIHSS as indicators, comprising a step of measuring IL-8 protein mass and NIHSS in a blood sample collected from a test animal.   The blood sample collected from the test animal is collected from the test animal at any time between the second day after the onset of cerebral infarction-like symptoms and the 20th day after the onset. The inspection method for predicting the prognosis of cerebral infarction according to claim 3.   A kit for cerebral infarction testing, comprising a reagent capable of measuring the protein amount of IL-8 protein.