Diagnosis of multiple sclerosis
The present invention relates to methods and kits for diagnosing multiple sclerosis (MS) in a subject. Particularly, the present invention relates to methods and kits for diagnosing a subtype of MS in a subject, the subtype selected from relapsing-remitting MS (RRMS), secondary progressive MS (SPMS), primary progressive MS (PPMS) and a pathologic sub-type of MS lesions selected from Pattern I and Pattern II MS lesion.
1. A method of diagnosing a subtype of multiple sclerosis (MS) in a subject, the method comprising determining the reactivity of antibodies in a sample obtained from the subject to a plurality of antigens selected from the group consisting of the antigens listed in Tables 1 to 4, thereby determining the reactivity pattern of the sample to the plurality of antigens, and comparing the reactivity pattern of said sample to a control reactivity pattern, wherein the subtype of MS is selected from the group consisting of:
(i) relapsing remitting multiple sclerosis (RRMS) wherein said plurality of antigens comprises at least 5 different antigens selected from the group consisting of the antigens listed in Table 1;
(ii) primary progressive multiple sclerosis (PPMS) wherein said plurality of antigens comprises at least 5 different antigens selected from the group consisting of the antigens listed in Table 2;
(iii) secondary progressive multiple sclerosis (SPMS) wherein said plurality of antigens comprises at least 5 different antigens selected from the group consisting of the antigens listed in Table 3; and
(iv) a pathologic subtype of MS selected form Pattern I lesions and Pattern II lesions wherein said plurality of antigens comprises at least 5 different antigens selected from the group consisting of the antigens listed in Table 4;
wherein said plurality of antigens is used in the form of an antigen array comprising a glass support, wherein determining the reactivity of antibodies in the sample to the plurality of antigens is performed using a system providing quantitative measurement of antigen-antibody binding by fluorescence detection, and wherein a significant difference between the reactivity pattern of said sample obtained from the subject compared to a reactivity pattern of a control sample is an indication that the subject is afflicted with the subtype of MS.
2. The method of claim 1 , wherein the glass support is an epoxy glass support.
3. The method of claim 2 , wherein each antigen is attached to at least three separate specific addressable locations of the array.
4. The method of claim 1 for diagnosing relapsing remitting multiple sclerosis (RRMS) in a subject, wherein the plurality of antigens are selected from the antigens listed in Table 1 and the control reactivity pattern is obtained from healthy subjects.
5. The method of claim 1 , wherein each antigen is attached to at least three separate specific addressable locations of the array.
6. The method of claim 4 , wherein the plurality of antigens comprises all the antigens listed in Table 1.
7. The method of claim 1 , for diagnosing primary progressive multiple sclerosis (PPMS) in a subject, wherein the plurality of antigens are selected from the antigens listed in Table 2 and the control reactivity pattern is obtained from healthy subjects.
8. The method of claim 7 , wherein the plurality of antigens comprises all the antigens listed in Table 2.
9. The method of claim 1 for diagnosing secondary progressive multiple sclerosis (SPMS) in a subject, wherein the plurality of antigens are selected from the antigens listed in Table 3 and the control reactivity pattern is obtained from RRMS subjects.
10. The method of claim 9 , wherein the plurality of antigens comprises all the antigens listed in Table 3.
11. The method of claim 1 for diagnosing Pattern I lesions in a subject with MS, wherein the plurality of antigens is selected from the antigens listed in Table 4 and the control reactivity pattern is obtained from subjects having Pattern II lesions.
12. The method of claim 1 for diagnosing Pattern II lesions in a subject with MS, wherein the plurality of antigens is selected from the antigens listed in Table 4 and the control reactivity pattern is obtained from subjects having Pattern I lesions.
13. The method of claim 1 , wherein the control is selected from the group consisting of a sample from at least one individual, a panel of control samples from a set of individuals, and a stored set of data from control individuals.
14. The method of claim 1 , wherein the sample is a serum sample.
15. The method of claim 1 , further comprising diluting the sample 1:10 before determining the reactivity of antibodies in the sample.
16. The method of claim 1 , wherein the subtype of MS is selected from the group consisting of:
(i) relapsing remitting multiple sclerosis (RRMS) wherein said plurality of antigens is HSP70 511-530, HSP60 286-305, LACTOCEREBROSIDE, AB1-42 and AB1-12, listed in Table 1;
(ii) primary progressive multiple sclerosis (PPMS) wherein said plurality of antigens is Chondroitin 4-Sulfate, Neurofilament 68 kDa, Amyloid Beta, Secreted APPalpha and SOD, listed in Table 2;
(iii) secondary progressive multiple sclerosis (SPMS) wherein said plurality of antigens is HSP60 376-395, Amyloid beta 1-23, HSP70 601-620, Cardiolipin and beta Crystallin, listed in Table 3; and
(iv) a pathologic subtype of MS selected from Pattern I lesions and Pattern II lesions wherein said plurality of antigens is 15-ketocholestane, 15α-hydroxycholestene, 15-ketocholestene, Brain L-α-lysophosphatidylserine and 160 kDa. Neurofilament, listed in Table 4.