Background
Methods
1. Patients
2. Analysis of CSF by WB for 14-3-3
3. Analysis of total tau protein in CSF samples
4. ELISA detection of 14-3-3 in CSF
5. Analysis by the real-time QUIC (RT-QUIC) method
6. Statistical analysis
7. MRI Protocol and PSWC on EEGs
Results
1. Detection of 14-3-3 by WB in CSF samples
14-3-3 γ-isoform | 14-3-3 all isoforms | ||
---|---|---|---|
ELISA | WB | WB | |
sensitivity (%) | 95.2 | 87.1 | 91.4 |
specificity (%) | 72.7 | 84.8 | 78.8 |
disease | number | male | female | 14-3-3 γ | 14-3-3 all isoforms | ||||
---|---|---|---|---|---|---|---|---|---|
ELISA | WB | WB | |||||||
average
| SD | min | max | ||||||
CJD | 124 | 69 | 55 | 263, 549 | 21, 525 | 135 | 75, 373 | 108/124 | 114/124 |
DAT | 54 | 33 | 21 | 1, 537 | 751.2 | 0 | 2, 410 | 3/54 | 7/54 |
CVD | 7 | 5 | 2 | 975.9 | 332.4 | 521 | 1, 51 2 | 0/7 | 2/7 |
PD | 5 | 4 | 1 | 542.2 | 20.3 | 521 | 565 | 0/5 | 0/5 |
PSP | 3 | 2 | 1 | 223.3 | 226.6 | 38 | 476 | 0/3 | 0/3 |
FTLD | 2 | 1 | 1 | 425 | 236.2 | 258 | 592 | 0/2 | 0/2 |
HD | 1 | 1 | 0 | 556 | 0. | 556 | 556 | 0/1 | 0/1 |
CBD | 2 | 2 | 0 | 449.5 | 245.4 | 276 | 623 | 0/2 | 0/2 |
ALS | 3 | 1 | 2 | 303.7 | 184.8 | 179 | 516 | 0/3 | 0/3 |
limbic encephalitis | 2 | 1 | 1 | 4440.5 | 1935.4 | 3, 072 | 5, 809 | 2/2 | 2/2 |
MELAS | 4 | 2 | 2 | 5, 069.5 | 222.7 | 4, 912 | 5, 227 | 4/4 | 4/4 |
PCD/LEMS | 2 | 1 | 1 | 4, 387 | 217.8 | 4, 233 | 4, 541 | 2/2 | 2/2 |
temporal epilepsy | 4 | 1 | 3 | 3, 356.8 | 1332.4 | 1, 530 | 4, 696 | 4/4 | 4/4 |
MCI | 3 | 1 | 2 | 603 | 20.8 | 591 | 627 | 0/3 | 0/3 |
Dementia, etiology unknown | 3 | 1 | 2 | 410.3 | 150.5 | 258 | 559 | 0/3 | 0/3 |
healthy subject | 4 | 2 | 2 | 0 | 0 | 0 | 0 | 0/4 | 0/4 |
2. Detection of total tau protein in CSF Samples
3. Detection of 14-3-3 in CSF samples by ELISA
4. Analysis of biochemical markers in 21 early-stage CJD patients
Age | Sex | CJD type | d.w. | CSF | MRI | ||||
---|---|---|---|---|---|---|---|---|---|
ELISA of 14-3-3 protein AU/ml | the detection of WB method of 14-3-3a protein | the detection of WB method of 14-3-3γ protein | Real-time QUIC | DWI | |||||
71 | m | sp | probable | 0 | 4, 157 | - | - | + | + |
77 | m | sp | probable | 2 | 10, 812 | + | + | + | + |
64 | f | sp | probable | 4 | 11, 812 | + | + | + | + |
73 | m | sp | probable | 4 | 3, 850 | + | + | - | + |
67 | m | sp | probable | 4 | 10, 814 | + | + | + | + |
76 | m | sp | probable | 4 | 6, 772 | + | + | + | + |
80 | f | sp | probable | 4 | 9, 850 | + | + | + | + |
63 | f | sp | probable | 4 | 2, 987 | - | - | - | + |
67 | m | sp | probable | 4 | 3, 553 | + | + | + | + |
70 | f | sp | probable | 4 | 3, 729 | - | - | - | + |
63 | m | sp | probable | 4 | 6, 133 | + | + | + | + |
63 | m | sp | probable | 4 | 8, 297 | + | + | + | + |
67 | f | sp | probable | 5 | 3, 372 | + | - | - | - |
74 | m | sp | probable | 5 | 7, 184 | + | + | + | + |
69 | f | sp | definite | 6 | 5, 600 | + | + | + | + |
54 | f | sp | definite | 6 | 1, 897 | - | - | - | + |
70 | f | ia | probable | 6 | 9, 460 | + | + | + | - |
70 | f | fa | probable | 6 | 4, 417 | + | + | + | + |
64 | f | sp | probable | 6 | 3, 888 | + | + | + | + |
51 | f | ia | probable | 6 | 6, 313 | + | + | + | + |
74 | f | sp | probable | 6 | 6, 133 | + | + | + | + |
5. Analysis of CJD patients where 14-3-3γ was not detected by WB yet the concentration of 14-3-3γ exceeded the ELISA cut-off level (Table 4)
Age | Sex | type | Polymorphism of codon129 in PRNP gene | Mutation of PRNP gene | ELISA of 14-3-3 protein (AU/ml) | Real-time QUIC |
---|---|---|---|---|---|---|
69 | male | sp | MM | - | 2, 101 | + |
33 | female | sp | MM | - | 1, 852 | + |
70 | female | sp | MM | - | 2, 452 | - |
73 | female | sp | MM | - | 2, 531 | - |
64 | male | sp | MM | - | 5, 493 | + |
65 | male | fa | MM | E200K | 4, 621 | + |
79 | female | sp | VV | - | 2, 342 | + |
6. Analysis of CJD patients where 14-3-3γ was not detected using WB and 14-3-3γ concentration was below the cut-off level for the ELISA (Table 5)
Age | Sex | type | Polymorphism of codon129 in PRNP gene | Mutation of PRNP gene | ELISA of 14-3-3 protein (AU/ml) | Real-time QUIC |
---|---|---|---|---|---|---|
85 | female | ge | MM | V180I | 1, 086 | - |
84 | female | ge | MM | V180I | 1, 222 | - |
84 | male | ge | MM | V180I | 1, 313 | - |
84 | female | ge | MM | V180I | 1, 383 | - |
75 | male | ge | MM | M232R | 1, 676 | + |
78 | male | sp | MM | - | 135 | - |
66 | male | sp | MM | - | 1, 224 | + |
7. Analysis of non-CJD patients with positive 14-3-3γ ELISA diagnoses (Table 6)
Discussion
Age | Sex | diagnosis | WB (14-3-3 protein) | ELISA | Real-time QUIC method | |
---|---|---|---|---|---|---|
all isoforms | γ-specific isoform | γ-specific isoform | ||||
79 | male | PCD/LEMS | + | + | 4, 233 | - |
69 | male | PCD/LEMS | + | + | 4, 541 | - |
51 | female | temporal epilepsy | + | + | 3, 405 | - |
59 | female | temporal epilepsy | + | + | 3, 796 | - |
62 | male | limbic encephalitis | + | + | 3, 072 | - |
78 | male | limbic encephalitis | + | + | 5, 809 | - |
42 | female | MELAS | + | + | 4, 912 | - |
25 | female | MELAS | + | + | 5, 227 | - |
78 | male | DAT | + | + | 3, 110 | - |
62 | female | DAT | + | + | 3, 386 | - |
69 | female | DAT | + | + | 3, 892 | - |
57 | male | DAT | + | - | 3, 092 | - |
59 | male | DAT | + | - | 3, 074 | - |
61 | male | DAT | + | - | 3, 038 | - |
66 | female | DAT | + | - | 2, 941 | - |
67 | female | DAT | - | - | 2, 569 | - |
70 | female | DAT | - | - | 2, 534 | - |
78 | female | DAT | - | - | 2, 445 | - |
83 | female | DAT | - | - | 2, 410 | - |
89 | female | DAT | - | - | 2, 025 | - |
72 | male | DAT | - | - | 1, 949 | - |
59 | female | DAT | - | - | 1, 852 | - |
63 | male | DAT | - | - | 1, 709 | - |