Limits...
Detection of CWD prions in urine and saliva of deer by transgenic mouse bioassay.

Haley NJ, Seelig DM, Zabel MD, Telling GC, Hoover EA - PLoS ONE (2009)

Bottom Line: The mechanisms of CWD transmission are poorly understood, though bodily fluids are thought to play an important role.In addition, PrP(CWD) was detected in pooled and concentrated urine by protein misfolding cyclic amplification (PMCA).These findings help extend our understanding of CWD prion shedding and transmission and portend the detection of infectious prions in body fluids in other prion infections.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.

ABSTRACT
Chronic wasting disease (CWD) is a prion disease affecting captive and free-ranging cervids (e.g. deer, elk, and moose). The mechanisms of CWD transmission are poorly understood, though bodily fluids are thought to play an important role. Here we report the presence of infectious prions in the urine and saliva of deer with chronic wasting disease (CWD). Prion infectivity was detected by bioassay of concentrated, dialyzed urine and saliva in transgenic mice expressing the cervid PrP gene (Tg[CerPrP] mice). In addition, PrP(CWD) was detected in pooled and concentrated urine by protein misfolding cyclic amplification (PMCA). The concentration of abnormal prion protein in bodily fluids was very low, as indicated by: undetectable PrP(CWD) levels by traditional assays (western blot, ELISA) and prolonged incubation periods and incomplete TSE attack rates in inoculated Tg(CerPrP) mice (373(+/-)3 days in 2 of 9 urine-inoculated mice and 342(+/-)109 days in 8 of 9 saliva-inoculated mice). These findings help extend our understanding of CWD prion shedding and transmission and portend the detection of infectious prions in body fluids in other prion infections.

Show MeSH

Related in: MedlinePlus

Western Blot detection of PrPCWD in urine and saliva-inoculated mice.Western blotting analysis of control and test mice, demonstrating PrPCWD in positive control mice (lanes 1 and 2), as well as urine (lanes 3 and 4) and saliva (lanes 5 and 6) inoculated mice. Protease-resistant prions were not detected in negative control mice (lanes 7 and 8). Flanking lanes represent undigested PrPC.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2654070&req=5

pone-0004848-g002: Western Blot detection of PrPCWD in urine and saliva-inoculated mice.Western blotting analysis of control and test mice, demonstrating PrPCWD in positive control mice (lanes 1 and 2), as well as urine (lanes 3 and 4) and saliva (lanes 5 and 6) inoculated mice. Protease-resistant prions were not detected in negative control mice (lanes 7 and 8). Flanking lanes represent undigested PrPC.

Mentions: In western blotting, PrPCWD proteinase K-resistant glycoforms spanned 21–27 kD. In all cases, the dominant PrPCWD glycoform was the di-glycosylated band, followed by mono- and non-glycosylated isoforms (Figure 2).


Detection of CWD prions in urine and saliva of deer by transgenic mouse bioassay.

Haley NJ, Seelig DM, Zabel MD, Telling GC, Hoover EA - PLoS ONE (2009)

Western Blot detection of PrPCWD in urine and saliva-inoculated mice.Western blotting analysis of control and test mice, demonstrating PrPCWD in positive control mice (lanes 1 and 2), as well as urine (lanes 3 and 4) and saliva (lanes 5 and 6) inoculated mice. Protease-resistant prions were not detected in negative control mice (lanes 7 and 8). Flanking lanes represent undigested PrPC.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2654070&req=5

pone-0004848-g002: Western Blot detection of PrPCWD in urine and saliva-inoculated mice.Western blotting analysis of control and test mice, demonstrating PrPCWD in positive control mice (lanes 1 and 2), as well as urine (lanes 3 and 4) and saliva (lanes 5 and 6) inoculated mice. Protease-resistant prions were not detected in negative control mice (lanes 7 and 8). Flanking lanes represent undigested PrPC.
Mentions: In western blotting, PrPCWD proteinase K-resistant glycoforms spanned 21–27 kD. In all cases, the dominant PrPCWD glycoform was the di-glycosylated band, followed by mono- and non-glycosylated isoforms (Figure 2).

Bottom Line: The mechanisms of CWD transmission are poorly understood, though bodily fluids are thought to play an important role.In addition, PrP(CWD) was detected in pooled and concentrated urine by protein misfolding cyclic amplification (PMCA).These findings help extend our understanding of CWD prion shedding and transmission and portend the detection of infectious prions in body fluids in other prion infections.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.

ABSTRACT
Chronic wasting disease (CWD) is a prion disease affecting captive and free-ranging cervids (e.g. deer, elk, and moose). The mechanisms of CWD transmission are poorly understood, though bodily fluids are thought to play an important role. Here we report the presence of infectious prions in the urine and saliva of deer with chronic wasting disease (CWD). Prion infectivity was detected by bioassay of concentrated, dialyzed urine and saliva in transgenic mice expressing the cervid PrP gene (Tg[CerPrP] mice). In addition, PrP(CWD) was detected in pooled and concentrated urine by protein misfolding cyclic amplification (PMCA). The concentration of abnormal prion protein in bodily fluids was very low, as indicated by: undetectable PrP(CWD) levels by traditional assays (western blot, ELISA) and prolonged incubation periods and incomplete TSE attack rates in inoculated Tg(CerPrP) mice (373(+/-)3 days in 2 of 9 urine-inoculated mice and 342(+/-)109 days in 8 of 9 saliva-inoculated mice). These findings help extend our understanding of CWD prion shedding and transmission and portend the detection of infectious prions in body fluids in other prion infections.

Show MeSH
Related in: MedlinePlus