NR AOUS
AU Kilidireas,C.; Detzortzis,N.; Muselimi,L.; Politi,A.; Kapaki,E.; Patsouris,E.; Davaki,P.; Vasilopoulos,D.
TI Quantitative Western Blot Analysis of CSF 14-3-3 Protein in CJD Patients and Other Neurologic Patients
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - DG-73
PT Konferenz-Poster
AB
Twenty seven patients with definite CJD were studied by Quantitative Western Blot analysis for CSF 14-3-3 concentration. Thirty six other Dementia patients were studied in parallel. Band identification was carried out as strongly immunoreactive 30 kd band to anti- 14-3-3946; sc antibody co-migrating with en equally immunoreactive 30 band of LAN-5 human neuroblastoma protein extracts. Quantitative Blot was developed by the logismic analysis of the optical density of identified 14-3-3 protein. LAN-5 human neuroblastoma extracts' identified 14-3-3 protein was used as standard.
Twenty sixe out of 27 CJD patients showed 14-3-3 concentration >2000 A.U. (2340-7870 Au). The other dementia patients showed either not presence, trace or weak immunoreactive bands corresponded to 410-1530 AU of 14-3-3 concentration, so the sensitivity of the assay is 96,39% and the specificity 100%. Two out of 26 definite CJD patients were initialy negative (weak band, concentration <2000 A.U.) and showed increase in CSF 14-3-3 concentration within in 3 months in repeated samples. 6 other dementia patients showed no increase in repeated samples. In addition three probable CJD patients who were initialy negative became positive within 3 months with increase in 14-3-3 concentration >2.000 AU.
All the patients and the controls who had repeated samples were tested for other 14-3-3 isoforms (950;, 949;, 952;, 947;, 963;) presented weak or not at all immunoreactivity and a significant increase was only in 946; isoform.
In conclusion a) Quantitative Blot increases the reliability of the assay b) repeated samples are important in case of suspect CJD patients and c the 946; isoform seems to be specifically increased in CJD patients among the other isoforms tested.
AD C. Kilidireas, N. Detzortzis, L. Muselimi, A. Politi, E. Kapaki, E. Patsouris, P. Davaki, D. Vasilopoulos, National and Kapodistrian University of Athens, Medical School, Department of Neurology, Athens, Greece
SP englisch
PO Deutschland