Diagnostic Method for Pediatric Acute-Onset Neuropsychiatric Syndrome (PANS) and Pediatric Autoimmune Neuropsychiatric Disorder Associated with Streptococci Infection (PANDAS)

Abstract

The present invention provides a panel of at least five clinical analyses or tests (using serum samples) to determine the risk of pediatric acute-onset neuropsychiatric syndrome (PANS) and/or pediatric autoimmune neuropsychiatric disorder associated with streptococcal infection (PANDAS) in an individual. These include enzyme linked immunosorbent assays (ELISAs) to measure antibody titers against neuronal antigens present in the brain; the neuronal antigens include lysoganglioside, tubulin, dopamine receptor D1, dopamine receptor D2, serotonin receptor 5HT2A, and serotonin receptor 5HT2C. Antibody titers against at least four of these neuronal antigens are required in the present methods; preferably antibody tiers against all of these neuronal antigens are measured. A final assay is used to quantify calcium/calmodulin-dependent protein kinase activity using a neuronal cell line. The results of these analyses or tests are then combined using an algorithm to determine whether a PANS or PANDAS diagnosis is appropriate for the individual. Depending on the diagnosis, an appropriate treatment can be determined.

1 . A method for determining the likelihood of a PANS or PANDAS diagnosis in a patient, said method comprising
 (1) obtaining a serum sample from the patient;   (2) in a first assay, measuring a first antibody titer against lysoganglioside in a first portion of the serum sample and comparing the measured first antibody titer to a first normal control titer against lysoganglioside;   (3) in a second assay, measuring a second antibody titer against tubulin in a second portion of the serum sample and comparing the measured second antibody titer to a second normal control titer against tubulin;   (4) in a third assay, measuring a third antibody titer against dopamine receptor D1 in a third portion of the serum sample and comparing the measured third antibody titer to a third normal control titer against dopamine receptor D1;   (5) in a fourth assay, measuring a fourth antibody titer against dopamine receptor D2 in a fourth portion of the serum sample and comparing the measured fourth antibody titer to a fourth normal control titer against dopamine receptor D2; and   (6) in a final assay, measuring calcium/calmodulin-dependent protein kinase activity using a neuronal cell line in a fifth portion of the serum sample and comparing the measured calcium/calmodulin-dependent protein kinase activity to a normal control calcium/calmodulin-dependent protein kinase activity;   whereby a PANS or PANDAS diagnosis is considered likely if any one of the measured first, second, third, or fourth antibody titers is significantly higher than the corresponding first, second, third, or fourth normal control titers, respectively; whereby the PANS or PANDAS diagnosis is considered highly likely if, independent of results from the first, second, third, and fourth assays, the measured calcium/calmodulin-dependent protein kinase activity is significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity; and whereby the PANS or PANDAS diagnosis is considered not likely if (1) none of the first, second, third, or fourth antibody titers are significantly higher than the corresponding first, second, third, or fourth normal control titers, respectively, and (2) the measured calcium/calmodulin-dependent protein kinase activity is not significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity.   
     
     
         2 . The method of  claim 1 , wherein the serum sample is obtained from whole blood from the patient. 
     
     
         3 . The method of  claim 2 , wherein measured antibody titer is considered significantly higher than the corresponding normal control titer if the measured antibody titer is greater than the normal control titer's mean value by at least 1.5 times the normal control titer's standard deviation; and wherein measured calcium/calmodulin-dependent protein kinase activity is considered significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity if the measured calcium/calmodulin-dependent protein kinase activity is greater than the normal calcium/calmodulin-dependent protein kinase activity's mean value by at least 2 times calcium/calmodulin-dependent protein kinase activity's standard deviation. 
     
     
         4 . The method of  claim 2 , wherein measured antibody titer is considered significantly higher than the corresponding normal control titer if the measured antibody titer is greater than the normal control titer's mean value by at least 2 times normal control titer's standard deviation; and wherein measured calcium/calmodulin-dependent protein kinase activity is considered significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity if the measured calcium/calmodulin-dependent protein kinase activity is greater than the normal calcium/calmodulin-dependent protein kinase activity's mean value by at least 3 times calcium/calmodulin-dependent protein kinase activity's standard deviation. 
     
     
         5 . A method for treating PANS or PANDAS in a patient, said method comprising (A) requesting an analysis of a serum sample from a patient suspected of having PANS or PANDAS, wherein the analysis is capable of determining whether the patient is likely or highly likely to have PANS or PANDAS and (B), should the analysis determine that the patient is likely or highly likely to have PANS or PANDAS, administering antibiotic therapy, anti-inflammatory therapy, intravenous immunoglobulin therapy, plasmaphoresis therapy, or combinations thereof to the patient;
 wherein the analysis comprises the steps of:   (1) obtaining a serum sample from the patient;   (2) in a first assay, measuring a first antibody titer against lysoganglioside in a first portion of the serum sample and comparing the measured first antibody titer to a first normal control titer against lysoganglioside;   (3) in a second assay, measuring a second antibody titer against tubulin in a second portion of the serum sample and comparing the measured second antibody titer to a second normal control titer against tubulin;   (4) in a third assay, measuring a third antibody titer against dopamine receptor D1 in a third portion of the serum sample and comparing the measured third antibody titer to a third normal control titer against dopamine receptor D1;   (5) in a fourth assay, measuring a fourth antibody titer against dopamine receptor D2 in a fourth portion of the serum sample and comparing the measured fourth antibody titer to a fourth normal control titer against dopamine receptor D2; and   (6) in a final assay, measuring calcium/calmodulin-dependent protein kinase activity using a neuronal cell line in a fifth portion of the serum sample and comparing the measured calcium/calmodulin-dependent protein kinase activity to a normal control calcium/calmodulin-dependent protein kinase activity;   whereby the patient is considered likely to have PANS or PANDAS if any one of the measured first, second, third, or fourth antibody titers is significantly higher than the corresponding first, second, third, or fourth normal control titers, respectively; whereby the patient is considered highly likely to have PANS or PANDAS if, independent of results from the first, second, third, and fourth assays, the measured calcium/calmodulin-dependent protein kinase activity is significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity; and whereby the patient is considered not likely to have PANS or PANDAS if (1) none of the first, second, third, or fourth antibody titers are significantly higher than the corresponding first, second, third, or fourth normal control titers, respectively, and (2) the measured calcium/calmodulin-dependent protein kinase activity is not significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity.   
     
     
         6 . The method of  claim 5 , wherein the serum sample is obtained from whole blood from the patient. 
     
     
         7 . The method of  claim 5 , wherein measured antibody titer is considered significantly higher than the corresponding normal control titer if the measured antibody titer is greater than the normal control titer's mean value by at least 1.5 times normal control titer's standard deviation; and wherein measured calcium/calmodulin-dependent protein kinase activity is considered significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity if the measured calcium/calmodulin-dependent protein kinase activity is greater than the normal calcium/calmodulin-dependent protein kinase activity's mean value by at least 2 times calcium/calmodulin-dependent protein kinase activity's standard deviation. 
     
     
         8 . The method of  claim 5 , wherein measured antibody titer is considered significantly higher than the corresponding normal control titer if the measured antibody titer is greater than the normal control titer's mean value by at least 2 times normal control titer's standard deviation; and wherein measured calcium/calmodulin-dependent protein kinase activity is considered significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity if the measured calcium/calmodulin-dependent protein kinase activity is greater than the normal calcium/calmodulin-dependent protein kinase activity's mean value by at least 3 times calcium/calmodulin-dependent protein kinase activity's standard deviation. 
     
     
         9 . A method for determining the likelihood of a PANS or PANDAS diagnosis in a patient, said method comprising
 (1) obtaining a serum sample from the patient;   (2) carrying out at least four assays on the serum sample, wherein the at least four assays are selected from the group consisting of
 (A) a first assay wherein a first antibody titer against lysoganglioside is measured in a first portion of the serum sample and then is compared to a first normal control titer against lysoganglioside, 
 (B) a second assay wherein a second antibody titer against tubulin is measured in a second portion of the serum sample and then is compared to a second normal control titer against tubulin, 
 (C) a third assay wherein a third antibody titer against dopamine receptor D1 is measured in a third portion of the serum sample and then is compared to a third normal control titer against dopamine receptor D1, 
 (D) a fourth assay wherein a fourth antibody titer against dopamine receptor D2 is measured in a fourth portion of the serum sample and then is compared to a fourth normal control titer against dopamine receptor D2, 
 (E) a fifth assay wherein a fifth antibody titer against human serotonin receptor 5HT2A is measured in a fifth portion of the serum sample and then is compared to a fifth normal control titer against human serotonin receptor 5HTT2A, and 
 (F) a sixth assay wherein a sixth antibody titer against human serotonin receptor 5HT2C is measured in a sixth portion of the serum sample and then is compared to a sixth normal control titer against human serotonin receptor 5HT2C; and 
   (3) carrying out a final assay wherein calcium/calmodulin-dependent protein kinase activity is measured using a neuronal cell line in a separate portion of the serum sample and then is compared to a normal control calcium/calmodulin-dependent protein kinase activity;   whereby a PANS or PANDAS diagnosis is considered likely if at least one of the measured first, second, third, fourth, fifth, or sixth antibody titers are significantly higher than the corresponding first, second, third, fourth, fifth, or sixth normal control titers, respectively; whereby the PANS or PANDAS diagnosis is considered highly likely if, independent of results from the first, second, third, fourth, fifth, or sixth assays, the measured calcium/calmodulin-dependent protein kinase activity is significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity; and whereby the PANS or PANDAS diagnosis is considered not likely if (1) none of the measured first, second, third, fourth, fifth, or sixth antibody titers are significantly higher than the corresponding first, second, third, fourth, fifth, or sixth normal control titers, respectively, and (2) the measured calcium/calmodulin-dependent protein kinase activity is not significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity.   
     
     
         10 . The method of  claim 9 , wherein the serum sample is obtained from whole blood from the patient. 
     
     
         11 . The method of  claim 10 , wherein measured antibody titer is considered significantly higher than the corresponding normal control titer if the measured antibody titer is greater than the normal control titer's mean value by at least 1.5 times the normal control titer's standard deviation; and wherein measured calcium/calmodulin-dependent protein kinase activity is considered significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity if the measured calcium/calmodulin-dependent protein kinase activity is greater than the normal calcium/calmodulin-dependent protein kinase activity's mean value by at least 2 times calcium/calmodulin-dependent protein kinase activity's standard deviation. 
     
     
         12 . The method of  claim 10 , wherein measured antibody titer is considered significantly higher than the corresponding normal control titer if the measured antibody titer is greater than the normal control titer's mean value by at least 2 times normal control titer's standard deviation; and wherein measured calcium/calmodulin-dependent protein kinase activity is considered significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity if the measured calcium/calmodulin-dependent protein kinase activity is greater than the normal calcium/calmodulin-dependent protein kinase activity's mean value by at least 3 times calcium/calmodulin-dependent protein kinase activity's standard deviation. 
     
     
         13 . The method of  claim 9 , wherein step (2) consists of carrying out at least five assays on the serum sample, wherein the at least five assays are selected from the group consisting of the first assay, the second assay, the third assay, the fourth assay, the fifth assay, and the sixth assay. 
     
     
         14 . The method of  claim 9 , wherein step (2) consists of carrying out all six assays of the group consisting of the first assay, the second assay, the third assay, the fourth assay, the fifth assay, and the sixth assay. 
     
     
         15 . A method for treating PANS or PANDAS in a patient, said method comprising (A) requesting an analysis of a serum sample from a patient suspected of having PANS or PANDAS, wherein the analysis is capable of determining whether the patient is likely or highly likely to have PANS or PANDAS and (B), should the analysis determine that the patient is likely or highly likely to have PANS or PANDAS, administering antibiotic therapy, anti-inflammatory therapy, intravenous immunoglobulin therapy, plasmaphoresis therapy, or combinations thereof to the patient;
 wherein the analysis comprises the steps of:   (1) obtaining the serum sample from the patient;   (2) carrying out at least four assays on the serum sample, wherein the at least four assays are selected from the group consisting of
 (A) a first assay wherein a first antibody titer against lysoganglioside is measured in a first portion of the serum sample and then is compared to a first normal control titer against lysoganglioside, 
 (B) a second assay wherein a second antibody titer against tubulin is measured in a second portion of the serum sample and then is compared to a second normal control titer against tubulin, 
 (C) a third assay wherein a third antibody titer against dopamine receptor D1 is measured in a third portion of the serum sample and then is compared to a third normal control titer against dopamine receptor D1, 
 (D) a fourth assay wherein a fourth antibody titer against dopamine receptor D2 is measured in a fourth portion of the serum sample and then is compared to a fourth normal control titer against dopamine receptor D2, 
 (E) a fifth assay wherein a fifth antibody titer against human serotonin receptor 5HT2A is measured in a fifth portion of the serum sample and then is compared to a fifth normal control titer against human serotonin receptor 5HTT2A, and 
 (F) a sixth assay wherein a sixth antibody titer against human serotonin receptor 5HT2C is measured in a sixth portion of the serum sample and then is compared to a sixth normal control titer against human serotonin receptor 5HT2C; and 
   (3) carrying out a final assay wherein calcium/calmodulin-dependent protein kinase activity is measured using a neuronal cell line in a separate portion of the serum sample and then is compared to a normal control calcium/calmodulin-dependent protein kinase activity;   whereby a PANS or PANDAS diagnosis is considered likely if at least one of the measured first, second, third, fourth, fifth, or sixth antibody titers are significantly higher than the corresponding first, second, third, fourth, fifth, or sixth normal control titers, respectively; whereby the PANS or PANDAS diagnosis is considered highly likely if, independent of results from the first, second, third, fourth, fifth, or sixth assays, the measured calcium/calmodulin-dependent protein kinase activity is significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity; and whereby the PANS or PANDAS diagnosis is considered not likely if (1) none of the measured first, second, third, fourth, fifth, or sixth antibody titers are significantly higher than the corresponding first, second, third, fourth, fifth, or sixth normal control titers, respectively, and (2) the measured calcium/calmodulin-dependent protein kinase activity is not significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity.   
     
     
         16 . The method of  claim 15 , wherein the serum sample is obtained from whole blood from the patient. 
     
     
         17 . The method of  claim 16 , wherein measured antibody titer is considered significantly higher than the corresponding normal control titer if the measured antibody titer is greater than the normal control titer's mean value by at least 1.5 times the normal control titer's standard deviation; and wherein measured calcium/calmodulin-dependent protein kinase activity is considered significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity if the measured calcium/calmodulin-dependent protein kinase activity is greater than the normal calcium/calmodulin-dependent protein kinase activity's mean value by at least 2 times calcium/calmodulin-dependent protein kinase activity's standard deviation. 
     
     
         18 . The method of  claim 16 , wherein measured antibody titer is considered significantly higher than the corresponding normal control titer if the measured antibody titer is greater than the normal control titer's mean value by at least 2 times normal control titer's standard deviation; and wherein measured calcium/calmodulin-dependent protein kinase activity is considered significantly higher than the normal control calcium/calmodulin-dependent protein kinase activity if the measured calcium/calmodulin-dependent protein kinase activity is greater than the normal calcium/calmodulin-dependent protein kinase activity's mean value by at least 3 times calcium/calmodulin-dependent protein kinase activity's standard deviation. 
     
     
         19 . The method of  claim 15 , wherein step (2) consists of carrying out at least five assays on the serum sample, wherein the at least five assays are selected from the group consisting of the first assay, the second assay, the third assay, the fourth assay, the fifth assay, and the sixth assay. 
     
     
         20 . The method of  claim 15 , wherein step (2) consists of carrying out all six assays of the group consisting of the first assay, the second assay, the third assay, the fourth assay, the fifth assay, and the sixth assay.