Abstract: Objective To establish the method of two-dimensional differential gel electrophoresis and obtain high resolution 2D images from cerebrospinal fluid （CSF） of patients with neurocysticercosis. Methods CSF samples were collected from four patients diagnosed as neurocysticercosis clinically and by ELISA, computed tomography （CT） or magnetic resonance imaging（MRI）, and from four healthy subjects without neurological disorders. The CSF samples were precipitated with cold acetone, then pooled by equal amount as patients and controls. The internal standard comprised equal amounts of proteins extracted from both groups. Internal standard，and proteins from the two groups were labeled prior to electrophoresis with spectrally resolvable fluorescent dyes，cyanin dye2（Cy2），Cy3 and Cy5. Sodium dodecylsulfonate polyacrylamide gel chromatography （SDS-PAGE） and two-dimensional differential in-gel electrophoresis （2-D DIGE） of labeled samples were then run. The differential expressed proteins showed in the images of SDS-PAGE and 2-D DIGE gels scanned with 488 nm，532 nm and 633 nm wavelength laser were analyzed by ImageQuant and DeCyde 5.0 respectively. Spot detection and quantification was performed for the differential in-gel analysis （DIA） module of DeCyder. Biological variation analysis （BVA） module of DeCyder was matched gel 1 and gel 2 images to provide data on differential protein expression levels between the two groups. Results The ImageQuant result displayed that the CSF protein was compatible with the dye，and the difference of protein amount was revealed by the difference of fluorescence intensity. DIA indicated that there were 896 and 894 protein dots on gel 1 and gel 2 respectively, and 90% of them were matched each other. BVA showed that there were 55 protein spots with different expressional level between neurocysticercosis and control groups. Protein spots with two-fold increase or decrease were 47 and 8 respectively in neurocysticercosis patients compared with healthy controls. Conclusion The method of 2-D DIGE has been established with high-resolution images for the examination of cerebrospinal fluid, providing a foundation for fur-ther study of neurocysticercosis comparative proteomics.

Key words: Neurocysticercosis, Cerebrospinal fluid, Proteomics, Differential gel electrophoresi