Detection of cerebrospinal fluid leakage by isoelectric focusing on polyacrylamide gels with silver staining using the PhastSystem?

Abstract

Cerebrospinal fluid (CSF) leakage, which sometimes occurs after skull trauma, is a life-threatening condition. A prompt start with antibiotics and/or prompt surgical treatment of fistulas is essential to avoid severe complications. This requires a fast and reliable method for detecting CSF leakage. This paper describes a fast (< 2 h) method based on the identification of the tau protein (beta 2-transferrin) band(s). Tau protein is a brain-specific variant of transferrin that is characteristic of CSF. The method includes iso-electric focusing (IEF) on pre-cast polyacrylamide gels and silver staining using the PhastSystem, an automated instrument for electrophoresis and staining. In the present study, this technology was applied on 200 consecutive CSF samples, 32 of which were from healthy volunteers. Tau protein was detected in all CSF samples but 5 (2.5%), all of which were from patients with blood-brain barrier (BBB) damage. In these cases, the tau protein band was indistinct when direct silver staining was used. Therefore, immunofixation with an antitransferrin antibody was performed, and after that the tau protein band was easy to detect. The specificity of the method was high, since no brain-specific tau protein band was detected in serum, tears, saliva, or nasal secretion. As IEF of CSF using the PhastSystem is increasingly used as the routine method for detection of oligoclonal bands of IgG in neurological disorders, it could readily be used in the clinical (neuro) chemical laboratory also for the less frequent cases of suspected CSF leakage.

Endonasal Endoscopic Closure of Cerebrospinal Fluid Rhinorrhea

“…Iatrogenic effractions during functional endoscopic sinus surgery (FESS) are more frequent in the anterior ethmoid region, due to variability of the thickness of the ethmoidal roof, increasing anteriorly to posteriorly. 9 Regions directly neighboring the lamina cribrosa are very fragile, in particular the internal hillside of the ethmoidal roof, 10 the main zone of fragility. Bony defects or dehiscences can occur in other regions, such as the tectal insertion of the middle turbinate, the canals of the anterior and posterior ethmoidal artery, dehiscent in 11 to 14% of cases.…”

Guidelines on routine cerebrospinal fluid analysis. Report from an EFNS task force

“…Isolelectric focusing uses a pH gradient to separate IgG populations on the basis of charge, which are then transferred onto a nitrocellulose or other membranes before immunostaining using an anti‐human immunoglobulin [39]. Some laboratories continue to use Sliver staining to detect oligoclonal bands (OCBs) with good results [7].…”

“…The Q alb is not influenced by intrathecal protein synthesis, is corrected for the plasma concentration of albumin, and is an integral part of intrathecal immunoglobulin synthesis formulae. The Q alb is a method‐independent measure, allowing the use of the same reference values in different laboratories [7,8]. However, there are no conclusive data on how the Q alb performs compared to total protein as a measure of blood–CSF barrier function in large cohorts of unselected patients.…”

Separation of β2-Transferrin by Denaturing Gel Electrophoresis to Detect Cerebrospinal Fluid in Ear and Nasal Fluids

“…Numerous studies have been published on this topic, some using polyacrylamide gels (2-4 ) or agarose gels (5, 6 ) combined with immunofixation (7,8 ) or with fluorescein (5 ), Coomassie Brilliant Blue (4 ), or silver staining (3 ). In addition to differences in band patterns, 2 proteins, prostaglandin-D synthase (␤-trace) and transthyretin (prealbumin), have been identified as possible specific constituents of CSF.…”