title: High sensitivity of an ELISA kit for detection of the gamma-isoform of 14-3-3 proteins: usefulness in laboratory diagnosis of human prion disease.
abstract: BACKGROUND
  The gamma-isoform of the 14-3-3 protein (14-3-3 gamma) is expressed in neurons, and could be a specific marker for neuronal damage. This protein has been reported as a detectable biomarker, especially in the cerebrospinal fluid (CSF) of Creutzfeldt-Jakob disease (CJD) patients by Western blotting (WB) or enzyme-linked immunosorbent assays (ELISAs). Western blotting for 14-3-3 gamma is not sensitive, and the reported data are conflicting among publications. An ELISA specific for 14-3-3 gamma is not available.
  
  
  METHODS
  CJD patients (n=114 sporadic CJD patients, 7 genetic CJD, and 3 iatrogenic CJD) and 99 patients with other neurodegenerative diseases were examined in this study. The CSF samples obtained were analyzed by Western blotting for 14-3-3 gamma, and by ELISA for total tau protein. We evaluated the sensitivity and specificity of the newly developed sandwich ELISA for 14-3-3 gamma.
  
  
  RESULTS
  The cut-off value of the 14-3-3 gamma ELISA was >1, 683 AU/ml; and sensitivity was 95.2%, with 72.7% specificity. This specificity was the same for the total tau protein ELISA. Seven CJD cases were negative by WB but positive using the 14-3-3 gamma ELISA, indicating that the ELISA is more sensitive. All 21 cases of early stage CJD could be diagnosed using a combination of the 14-3-3γ ELISA and diffusion weighted MR imaging (DWI-MRI).
  
  
  CONCLUSIONS
  The 14-3-3 gamma ELISA was more sensitive than conventional WB, and was useful for laboratory diagnosis of CJD, similar to the ELISA for the tau protein. Using DWI-MRI and these ELISA tests on CSF, diagnosis of CJD will be possible even at early stages of the disease.
authors.affiliation: Department of Pharmaceutical Care and Health Sciences, Faculty of Pharmaceutical Sciences, Fukuoka University, Fukuoka, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Matsui
authors.forename: Katsuya
authors.initials: K
authors.lastname: Satoh
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Miyazaki
authors.forename: Susumu
authors.initials: S
authors.lastname: Shirabe
authors.forename: Ryuichiro
authors.initials: R
authors.lastname: Atarashi
authors.forename: Kazuo
authors.initials: K
authors.lastname: Mutsukura
authors.forename: Akira
authors.initials: A
authors.lastname: Satoh
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Kataoka
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Nishida
mesh_terms: D048948:14-3-3 Proteins / Q000134:cerebrospinal fluid*; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000134:cerebrospinal fluid; D015153:Blotting, Western / Q000379:methods / Q000706:statistics & numerical data; D007562:Creutzfeldt-Jakob Syndrome / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D038524:Diffusion Magnetic Resonance Imaging / Q000379:methods; D042241:Early Diagnosis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000706:statistics & numerical data; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D019636:Neurodegenerative Diseases / Q000134:cerebrospinal fluid*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D016875:tau Proteins / Q000134:cerebrospinal fluid
chemical_list: D048948:14-3-3 Proteins; D015415:Biomarkers; D016875:tau Proteins