Hydrated autoclave pretreatment enhancement of prion protein immunoreactivity in formalin-fixed bovine spongiform encephalopathy-affected brain

Abstract

The efficacy of three pretreatment techniques for the detection of prion protein (PrP) in formalin-fixed, paraffin-embedded bovine spongiform encephalopathy (BSE)--affected brain tissue were compared using automated image analysis. The most abundant immunostaining was in the form of particulate expression observed in sections pretreated with hydrated autoclaving for 30 min. Considerably less immunostaining occurred in sections pretreated with formic acid and no specific particulate immunostaining was detected in sections pretreated with hydrolytic autoclaving. Hydrated autoclaving pretreatment of sections prior to PrP immunolabelling gives visualisation of widespread sites of abnormal PrP deposition in the brain, allowing detailed study of the form and distribution of the protein in routinely fixed bovine central nervous system affected with BSE.

Scrapie e seu diagnóstico diferencial em ovinos no Mato Grosso do Sul

“…Em ambos os animais, estas lesões, apesar de mais evidentes no tronco encefálico ao nível do óbex, foram observadas também no mesencéfalo, ponte, medula oblonga, cornos laterais e ventrais da medula espinhal, concordando com as descrições de Summers et al (1995), Driemeier (2007), Maxie & Youssef (2007) Hoinville (1996), este teste é de grande utilidade para o diagnóstico da doença em animais recém-infectados, em fase de incubação ou com sinais iniciais da doença, quando as alterações histopatológicas ainda podem estar ausentes ou serem incipientes, com possibilidade de diagnóstico falso negativo na histopatologia. Neste caso, segundo Haritani et al (1994) (Thomzig et al 2007), leite (Konold et al 2008, Lacroux et al 2008, Maddison et al 2009, Ligios et al 2011, fezes (Tamgüney 2009;,Terry et al 2011), urina (Murayama et al 2007, Gregori et al 2008, Rubenstein et al 2011, saliva (Vascellari et al 2007, sangue (Murayama et al 2007, Edwards et al 2010 e secreções nasais (Bessen et al 2010). A placenta, segundo é via por onde o PrP sc é mais intensamente eliminado, mas as demais fontes não devem ser negligenciadas, ao se estabelecer medidas de controle sanitário da doença.…”

Detection of the Disease-Associated Isoform of the Prion Protein in Formalin-Fixed Tissues by Western Blot

“…IHC does not depend upon proteinase K for removal of PrP C , however, it does require significant training and expertise for reliable differentiation of positive and negative cases. 2,7 Each approach has advantages and disadvantages, and it is preferred that more than 1 technique (typically both IHC and Western blot) would be applied to a given case before a final diagnosis is made. 4 Confounding the combined use of IHC and Western blot for a complete diagnosis of a TSE is the fact that Western-blot approaches require either fresh or frozen tissue whereas optimal conditions for IHC are formalin-fixed tissue that has never been frozen.…”

Metabolic disorders with clinical and radiologic features of sporadic Creutzfeldt-Jakob disease

“…Furthermore, 3F4 antibody staining by hydrolytic autoclaving method revealed no evidence of prions. 7,8 The findings were not considered consistent with prion disease. …”