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Technical Advance |
From the Pathophysiology Department,* Institute forNeuroscience, Tongji Medical School, Huazhong University of Science andTechnology, Wuhan, People’s Republic of China; and the Department ofNeurochemistry, New York State Institute forBasic Research in Developmental Disabilities, Staten Island, New York
We have developed an ultrasensitive bienzyme-substrate-recycle
enzyme-linked immunosorbent assay for the measurement of Alzheimer’s
disease (AD) abnormally hyperphosphorylated tau in cerebrospinal fluid
(CSF). The assay, which recognizes attomolar amounts of
tau, is 
400 and 1300 times more sensitive than
conventional enzyme-linked immunosorbent assay in determining the
hyperphosphorylated tau and total tau, respectively. With this
method, we measured both total tau and tau phosphorylated at
Ser-396/Ser-404 in lumbar CSFs from AD and control patients. We found
that the total tau was 215 ± 77 pg/ml in cognitively normal
control (n = 56), 234 ± 92 pg/ml in
non-AD neurological (n = 37), 304 ±
126 pg/ml in vascular dementia (n = 46),
and 486 ± 168 pg/ml (n = 52) in AD
patients, respectively. However, a remarkably elevated
level in phosphorylated tau was only found in AD (187 ± 84
pg/ml), as compared with normal controls (54 ± 33
pg/ml), non-AD (63 ± 34 pg/ml), and vascular
dementia (72 ± 33 pg/ml) groups. If we used the ratio of
hyperphosphorylated tau to total tau of 
0.33 as cutoff for AD
diagnosis, we could confirm the diagnosis in 96% of the
clinically diagnosed patients with a specificity of 95%,
86%, 100%, and 94% against nonneurological,
non-AD neurological, vascular dementia, and all of the
three control groups combined, respectively. It is suggested
that the CSF level of tau phosphorylated at Ser-396/Ser-404 is a
promising diagnostic marker of AD.
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