NR AWJC
AU Lipscomb,I.P.; Harris,K.; Pinchin,H.; Collin,R.; Keevil,C.W.
TI Dual staining of prion amyloid and general proteinaceous contamination on surgical steel surfaces enhances prion-positive signal differentiation
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions SA-10
PT Konferenz-Poster
AB Transmissible spongiform encephalopathies are a group of rare, transmissible, and fatal neurodegenerative diseases associated with the protein agent (PrPsc). As such, the sensitive and rapid detection of prion PrPsc amyloid on the surface of suspect surgical instruments is of great importance and may even allow remedial action to be taken by sterile service departments (SSDs) prior to any further operative intervention and possible iatrogenic transmission. However, conventional PrPsc detection methodologies tend to rely on the inefficient and unreliable removal of suspect material from a surface using swabs or wipes prior to antibody analysis. Here we show how the combination of an advanced light microscope technique, Episcopic Differential Interference Contrast / Epi-Fluorescence (EDIC/EF) microscopy, and the combination of fluorescent dyes (Sypro Ruby and thiazole derivatives) can be used to detect, in-situ, sub-micron (attomole) levels of both general and PrPsc protein contamination in brain and spleen sections, smears, and homogenate on surgical stainless steel surfaces and surgical instruments. This technique can be used to verify that surgical instruments are substantially free from prion protein soiling and hence reduce the risk of iatrogenic transmission. This technique was used to screen the contamination of instrument sets obtained from health service trusts and also assess decontamination protocols for PrPsc infected brain material dried on stainless steel surfaces. The results indicate that drying times in excess of 10 minutes and different detergent or enzymatic cleaning chemistries significantly alter the ability to remove PrPsc from the surfaces. This supports the requirement to prevent instrument drying before processing in SSDs, irrespective of the ultimate cleaning chemistry.
AD School of Biological Sciences, University of Southampton, Southampton, SO16 7PX, UK. E-mail: i.lipscomb@soton.ac.uk
SP englisch
PO Italien