Decontamination of surgical instruments from prion proteins: in vitro studies on the detachment, destabilization and degradation of PrPSc bound to steel surfaces.
Effective reprocessing of surgical instruments ensuring elimination of inadvertent contamination with infectious agents causing transmissible spongiform encephalopathies (TSEs) is essential for the prevention of iatrogenic transmission of Creutzfeldt-Jakob disease (CJD) or its new variant (vCJD) from asymptomatic carriers. In a search for effective yet instrument-friendly and routinely applicable reprocessing procedures, we used an in vitro carrier assay to assess the decontamination activity exerted by different reagents on pathological prion protein (PrP(Sc)), the biochemical marker for TSE infectivity, attached to steel surfaces. In this assay, steel wires were contaminated with 263K scrapie brain homogenate and reprocessed for decontamination by exposure to several different test reagents. Residual contamination with PrP(Sc) and its protease-resistant core PrP27-30, still present after reprocessing on the wire surface or in the cleaning solution, was monitored by sensitive Western blot detection without or after proteinase K digestion. Using this approach, various reagents and processing conditions were screened for both their efficacy of decontamination and their active principles, such as detachment, destabilization or degradation of surface-bound prion protein. This revealed that, under appropriate conditions, relatively mild reagents such as 0.2 % SDS/0.3 % NaOH (pH 12.8), a commercially available alkaline cleaner (pH 11.9-12.2), a disinfectant containing 0.2 % peracetic acid and low concentrations of NaOH (pH 8.9) or 5 % SDS (pH 7.1) exert potent decontaminating activities on PrP(Sc)/PrP27-30 attached to steel surfaces. For in vivo validation, wires reprocessed in these reagents have been implanted into reporter animals in ongoing experiments.
Lemmer
Karin
K
P24 - Transmissible spongiforme Enzephalopathien, Robert Koch-Institut, Nordufer 20, 13353 Berlin, Germany.
Mielke
Martin
M
FG 14 - Angewandte Krankenhaushygiene und Infektionsprävention, Robert Koch-Institut, Nordufer 20, 13353 Berlin, Germany.
Pauli
Georg
G
ZBS1 - Hochpathogene virale Erreger, Robert Koch-Institut, Nordufer 20, 13353 Berlin, Germany.
Beekes
Michael
M
P24 - Transmissible spongiforme Enzephalopathien, Robert Koch-Institut, Nordufer 20, 13353 Berlin, Germany.
D003666:Decontamination / Q000379:methods*; D006146:Guanidines / Q000494:pharmacology; D018620:PrPSc Proteins / Q000302:isolation & purification* / Q000378:metabolism; D012972:Sodium Hydroxide / Q000494:pharmacology; D012973:Sodium Hypochlorite / Q000494:pharmacology; D013232:Steel; D013525:Surgical Instruments; D013861:Thiocyanates / Q000494:pharmacology; D014508:Urea / Q000494:pharmacology
D006146:Guanidines; D018620:PrPSc Proteins; D013861:Thiocyanates; D013232:Steel; D012972:Sodium Hydroxide; C054436:guanidine thiocyanate; D014508:Urea; D012973:Sodium Hypochlorite