Blood-based screen for detecting neurological diseases in primary care settings

Abstract

The present invention includes methods and kits for the diagnosing a neurological disease within primary care settings comprising: obtaining a blood test sample from a subject, measuring IL-7 and TNFα biomarkers in the blood sample, comparing the level of the one or a combination of biomarkers and neurocognitive screening tests with the level of a corresponding one or combination of biomarkers in a normal blood sample and neurocognitive screening tests, and predicting that an increase in the level of the blood test sample in relation to that of the normal blood sample indicates that the subject is likely to have a neurological disease.

1 . A method of screening for neurological disease comprising:
 obtaining a blood test sample from a subject;   measuring two or more biomarkers in the blood sample selected from IL7, TNFα, IL5, IL6, CRP, IL10, TNC, ICAM1, FVII, 1309, TNFR1, A2M, TARC, eotaxin3, VCAM1, TPO, FABP, IL18, B2M, SAA, PPY, DJ1, and α-synuclein;   comparing the levels of the measured biomarkers with the levels of corresponding biomarkers in a normal blood sample;   determining that the subject is likely to have a neurological disease when the level of one or more of the measured biomarkers is increased as compared to the corresponding biomarker in the normal blood sample;   when the subject is determined as likely having a neurological disease, predicting the neurological disease based on the levels of the measured biomarkers; and   selecting a course of treatment for the subject based on the neurological disease predicted.   
     
     
         2 . The method of  claim 1 , wherein at least one of the biomarker measurements is obtained by a method selected from the group consisting of immunoassay and enzymatic activity assay. 
     
     
         3 - 4 . (canceled) 
     
     
         5 . The method of  claim 1 , wherein predicting the neurological disease includes using the levels of at least 3 of the measured biomarkers to distinguish between neurological diseases. 
     
     
         6 . The method of  claim 1 , wherein the isolated biological sample is serum or plasma. 
     
     
         7 . (canceled) 
     
     
         8 . The method of  claim 1 , wherein the neurological diseases are selected from Alzheimer's Disease, Parkinson's Disease, Down's syndrome, Frontotemporal dementia, Dementia with Lewy Bodies, and neurodegenerative disease. 
     
     
         9 . The method of  claim 1 , further comprising:
 determining the subject's age, and
 determining the subject's neurocognitive screening test results, 
 wherein the combination of two or more of the biomarkers, the subject's age, and the neurocognitive screening test results are at least 90% accurate in a primary care setting for the determination of Alzheimer's disease when compared to a control subject that does not have a neurological disease or disorder. 
   
     
     
         10 . The method of  claim 1 , wherein the step of predicting the neurological disease further comprises the step of determining one or more of the following parameters: sleep disturbance (yes/no), visual hallucinations (yes/no), psychiatric/personality changes (yes/no), age, and neurocognitive screening. 
     
     
         11 . The method of  claim 1 , wherein the level of expression of the two or more biomarkers is measured by at least one of fluorescence detection, chemiluminescence detection, electrochemiluminescence detection and patterned arrays, reverse transcriptase-polymerase chain reaction, antibody binding, fluorescence activated sorting, detectable bead sorting, antibody arrays, microarrays, enzymatic arrays, receptor binding arrays, allele specific primer extension, target specific primer extension, solid-phase binding arrays, liquid phase binding arrays, fluorescent resonance transfer, or radioactive labeling. 
     
     
         12 . The method of  claim 1 , wherein the method is used to screen for at least one of mild AD with an overall accuracy of at least 94% or very early AD with an overall accuracy of at least 91%. 
     
     
         13 . The method of  claim 1 , wherein:
 the specificity is in the range of 0.97 to 1.0;   the sensitivity is in the range of 0.80 to 1.0; and   the specificity is higher than the sensitivity.   
     
     
         14 - 25 . (canceled) 
     
     
         26 . A method of performing a clinical trial to evaluate a candidate drug believed to be useful in treating a neurological disease, the method comprising:
 (a) measuring at least two biomarkers selected from IL7, TNFα, IL5, IL6, CRP, IL10, TNC, ICAM1, FVII, 1309, TNFR1, A2M, TARC, eotaxin3, VCAM1, TPO, FABP, IL18, B2M, SAA, PPY, DJ1, and α-synuclein from one or more blood samples obtained from patients suspected of having a neurological disease, the patient's age, and results from one or more neurocognitive screening tests of the patient;   (b) administering a candidate drug to a first subset of the patients, and a placebo to a second subset of the patients;   (c) repeating step (a) after the administration of the candidate drug or the placebo; and   (d) determining if there is a statistically significant reduction in the expression of the one or more biomarkers in the first subset of the patients as compared to the second subset of patients,   wherein a statistically significant reduction indicates that the candidate drug is useful in treating the neurological disease.   
     
     
         27 . The method of  claim 26 , further comprising the steps of obtaining one or more additional blood samples from the patient after a predetermined amount of time and comparing the levels of the biomarkers from the one or more additional samples to determine disease progression. 
     
     
         28 . The method of  claim 26 , further comprising the steps of treating the patient for a pre-determined period of time, obtaining one or more additional blood samples from the patient after the predetermined amount of time and comparing the levels of the biomarkers from the one or more additional samples to determine disease progression. 
     
     
         29 . (canceled) 
     
     
         30 . A method of evaluating the effect of a treatment for a neurological disease, the method comprising:
 treating a patient for a neurological disease;   measuring an expression level of at least two biomarkers from blood samples obtained from patients suspected of having a neurological disease, the at least two biomarkers being selected from IL7, TNFα, IL5, IL6, CRP, IL10, TNC, ICAM1, FVII, 1309, TNFR1, A2M, TARC, eotaxin3, VCAM1, TPO, FABP, IL18, B2M, SAA, PPY, DJ1, and α-synuclein;   determining if there is a statistically significant reduction in the expression of one or more of the biomarkers in the patient following treatment for the neurological disease as compared to the expression level of corresponding biomarkers in a control;   wherein the control includes a second subset of patients that have not been treated or a prior sample obtained from the patient; and   wherein a statistically significant reduction indicates that the treatment is useful in treating the neurological disease.   
     
     
         31 . The method of  claim 1 , wherein the two or more biomarkers include at least IL-7 and TNFα. 
     
     
         32 . The method of  claim 31 , wherein elevated levels of expression of IL-7 and TNF alpha in the blood sample from the subject, as compared to the normal blood sample, indicates a greater likelihood that the subject suffers from the neurological disease. 
     
     
         33 . The method of  claim 1 , wherein the two or more biomarkers include at least CRP and IL10. 
     
     
         34 . (canceled) 
     
     
         35 . The method of  claim 33 , wherein reduced levels of expression of CRP and IL10 in the blood sample from the subject, as compared to the normal blood sample, indicates a greater likelihood that the individual suffers from the neurological disease. 
     
     
         36 . The method of claim_, further comprising the steps of obtaining one or more additional blood samples from the patient after a predetermined amount of time and comparing the levels of the biomarkers from the one or more additional samples to determine disease progression. 
     
     
         37 - 40 . (canceled) 
     
     
         41 . A kit for screening a subject for a neurological disease, the kit comprising:
 one or more reagents for detecting, in a blood sample obtained from the subject, an expression level of at least two biomarkers selected from IL-7, TNFα, IL5, IL6, CRP, IL10, TNC, ICAM1, FVII, 1309, TNFR1, A2M, TARC, eotaxin3, VCAM1, TPO, FABP, IL18, B2M, SAA, PPY, DJ1, and α-synuclein; and   instructions for comparing the measured expression levels of the at least two biomarkers in the blood sample obtained from the subject to a reference level;   wherein the reference level of each biomarker comprises a normalized measured level of the biomarker from one or more blood samples of human individuals without neurological disease.   
     
     
         42 . The kit of  claim 41 , wherein elevated levels of expression of IL-7 and TNF alpha in the blood sample obtained from the subject as compared to the reference level, indicates a greater likelihood that the subject suffers from the neurological disease. 
     
     
         43 . (canceled) 
     
     
         44 . The kit of  claim 41 , wherein reduced levels of expression of CRP and IL10 as compared to the reference level, indicates a greater likelihood that the individual suffers from the neurological disease. 
     
     
         45 . The kit of  claim 41 , wherein the sample is a serum sample. 
     
     
         46 . The kit of  claim 41 , wherein the neurological disease is selected from Alzheimer's Disease, Down's syndrome, Frontotemporal dementia, Dementia with Lewy Bodies, Parkinson's Disease, and dementia. 
     
     
         47 . The kit of  claim 41 , wherein the expression level of the biomarkers is measured through at least one of nucleic acid level, protein level, or functionally at the protein level. 
     
     
         48 . The kit of  claim 41 , wherein the expression level of the biomarkers is measured by at least one of fluorescence detection, chemiluminescence detection, electrochemiluminescence detection and patterned arrays, reverse transcriptase-polymerase chain reaction, antibody binding, fluorescence activated sorting, detectable bead sorting, antibody arrays, microarrays, enzymatic arrays, receptor binding arrays, allele specific primer extension, target specific primer extension, solid-phase binding arrays, liquid phase binding arrays, fluorescent resonance transfer, or radioactive labeling. 
     
     
         49 - 52 . (canceled)