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In vitro detection of prionemia in TSE-infected cervids and hamsters.

Elder AM, Henderson DM, Nalls AV, Wilham JM, Caughey BW, Hoover EA, Kincaid AE, Bartz JC, Mathiason CK - PLoS ONE (2013)

Bottom Line: Thus, it is important to monitor cervid and human blood products to ensure herd health and human safety.Current methods for detecting blood-associated prions rely primarily upon bioassay in laboratory animals.Our results indicate that RT-QuIC methodology as modified can provide consistent and reliable detection of blood-borne prions in preclinical and symptomatic stages of two animal TSEs, offering promise for prionemia detection in other species, including humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, United States of America.

ABSTRACT
Blood-borne transmission of infectious prions during the symptomatic and asymptomatic stages of disease occurs for both human and animal transmissible spongiform encephalopathies (TSEs). The geographical distribution of the cervid TSE, chronic wasting disease (CWD), continues to spread across North America and the prospective number of individuals harboring an asymptomatic infection of human variant Creutzfeldt-Jakob Disease (vCJD) in the United Kingdom has been projected to be ~1 in 3000 residents. Thus, it is important to monitor cervid and human blood products to ensure herd health and human safety. Current methods for detecting blood-associated prions rely primarily upon bioassay in laboratory animals. While bioassay provides high sensitivity and specificity, it requires many months, animals, and it is costly. Here we report modification of the real time quaking-induced conversion (RT-QuIC) assay to detect blood-borne prions in whole blood from prion-infected preclinical white-tailed deer, muntjac deer, and Syrian hamsters, attaining sensitivity of >90% while maintaining 100% specificity. Our results indicate that RT-QuIC methodology as modified can provide consistent and reliable detection of blood-borne prions in preclinical and symptomatic stages of two animal TSEs, offering promise for prionemia detection in other species, including humans.

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Related in: MedlinePlus

PrPD detection in hamster, white-tailed deer and muntjac by IHC.PrPD immunoreactivity in a spinal cord tissue section from a hamster 16 weeks after extranasal inoculation with HY-TME (A) detected with antibody 3F4 and ABC solution. PrPD immunoreactivity in the brainstem of CWD-infected white-tailed deer (C) and muntjac (E) detected with antibody BAR224 and AEC substrate. No immunoreactivity was seen in the corresponding tissues of mock-inoculated controls (B, D and F). The boxed areas are enlarged 10x in the insets. Scale bar = 200µm.
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pone-0080203-g007: PrPD detection in hamster, white-tailed deer and muntjac by IHC.PrPD immunoreactivity in a spinal cord tissue section from a hamster 16 weeks after extranasal inoculation with HY-TME (A) detected with antibody 3F4 and ABC solution. PrPD immunoreactivity in the brainstem of CWD-infected white-tailed deer (C) and muntjac (E) detected with antibody BAR224 and AEC substrate. No immunoreactivity was seen in the corresponding tissues of mock-inoculated controls (B, D and F). The boxed areas are enlarged 10x in the insets. Scale bar = 200µm.

Mentions: Immunohistochemistry was applied as a confirmation for the presence of PrPD deposition in the brains of animals where PrPC-converting activity was detected in blood. IHC was performed on both cervid and hamster TME-inoculated and mock-inoculated brains for detection of the disease associated isoform of the prion protein, PrPD. PrPD deposition was observed in TSE-infected animals, but not in mock-inoculated animals (Figure 7; Tables 1, 2).


In vitro detection of prionemia in TSE-infected cervids and hamsters.

Elder AM, Henderson DM, Nalls AV, Wilham JM, Caughey BW, Hoover EA, Kincaid AE, Bartz JC, Mathiason CK - PLoS ONE (2013)

PrPD detection in hamster, white-tailed deer and muntjac by IHC.PrPD immunoreactivity in a spinal cord tissue section from a hamster 16 weeks after extranasal inoculation with HY-TME (A) detected with antibody 3F4 and ABC solution. PrPD immunoreactivity in the brainstem of CWD-infected white-tailed deer (C) and muntjac (E) detected with antibody BAR224 and AEC substrate. No immunoreactivity was seen in the corresponding tissues of mock-inoculated controls (B, D and F). The boxed areas are enlarged 10x in the insets. Scale bar = 200µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815098&req=5

pone-0080203-g007: PrPD detection in hamster, white-tailed deer and muntjac by IHC.PrPD immunoreactivity in a spinal cord tissue section from a hamster 16 weeks after extranasal inoculation with HY-TME (A) detected with antibody 3F4 and ABC solution. PrPD immunoreactivity in the brainstem of CWD-infected white-tailed deer (C) and muntjac (E) detected with antibody BAR224 and AEC substrate. No immunoreactivity was seen in the corresponding tissues of mock-inoculated controls (B, D and F). The boxed areas are enlarged 10x in the insets. Scale bar = 200µm.
Mentions: Immunohistochemistry was applied as a confirmation for the presence of PrPD deposition in the brains of animals where PrPC-converting activity was detected in blood. IHC was performed on both cervid and hamster TME-inoculated and mock-inoculated brains for detection of the disease associated isoform of the prion protein, PrPD. PrPD deposition was observed in TSE-infected animals, but not in mock-inoculated animals (Figure 7; Tables 1, 2).

Bottom Line: Thus, it is important to monitor cervid and human blood products to ensure herd health and human safety.Current methods for detecting blood-associated prions rely primarily upon bioassay in laboratory animals.Our results indicate that RT-QuIC methodology as modified can provide consistent and reliable detection of blood-borne prions in preclinical and symptomatic stages of two animal TSEs, offering promise for prionemia detection in other species, including humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, United States of America.

ABSTRACT
Blood-borne transmission of infectious prions during the symptomatic and asymptomatic stages of disease occurs for both human and animal transmissible spongiform encephalopathies (TSEs). The geographical distribution of the cervid TSE, chronic wasting disease (CWD), continues to spread across North America and the prospective number of individuals harboring an asymptomatic infection of human variant Creutzfeldt-Jakob Disease (vCJD) in the United Kingdom has been projected to be ~1 in 3000 residents. Thus, it is important to monitor cervid and human blood products to ensure herd health and human safety. Current methods for detecting blood-associated prions rely primarily upon bioassay in laboratory animals. While bioassay provides high sensitivity and specificity, it requires many months, animals, and it is costly. Here we report modification of the real time quaking-induced conversion (RT-QuIC) assay to detect blood-borne prions in whole blood from prion-infected preclinical white-tailed deer, muntjac deer, and Syrian hamsters, attaining sensitivity of >90% while maintaining 100% specificity. Our results indicate that RT-QuIC methodology as modified can provide consistent and reliable detection of blood-borne prions in preclinical and symptomatic stages of two animal TSEs, offering promise for prionemia detection in other species, including humans.

Show MeSH
Related in: MedlinePlus