<i>PRNP</i> variants in goats reduce sensitivity of detection of PrP<sup>Sc</sup> by immunoassay

Madsen–Bouterse, Sally A.; Schneider, David A.; Dassanayake, Rohana P.; Truscott, Thomas C.; Zhuang, Dongyue; Kumpula-McWhirter, Nancy; O’Rourke, Katherine I.

doi:10.1177/1040638715585865

Cited by 10 publications

(19 citation statements)

References 38 publications

(89 reference statements)

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“…In addition, the performance of these tests for the identification of atypical scrapie and BSE in sheep was also evaluated. [10][11][12][13][14][15][16][17][18][19] Only IDEXX, a Bio-Rad SAP, b and Bio-Rad SG c rapid tests (enzyme-linked immunosorbent assays [ELISAs]) were able to detect atypical scrapie, a result also confirmed by routine screening for scrapie in sheep and goats. 3,22 In 2012, the European Food Safety Authority also recommended the PrioSTRIP SR d test (visual reading protocol) for the detection of TSE disease in small ruminants.…”

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confidence: 99%

EU-approved rapid tests might underestimate bovine spongiform encephalopathy infection in goats

et al. 2017

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We report the diagnostic sensitivity of 3 EU-approved rapid tests (ELISAs; 1 from IDEXX and 2 from Bio-Rad) for the detection of transmissible spongiform encephalopathy diseases in goats. Ninety-eight goat brainstem samples were tested. All the rapid tests had 100% specificity and ≥80% sensitivity, with the IDEXX test significantly more sensitive than the 2 Bio-Rad tests. All tests detected 100% of samples from goats with clinical scrapie, but missed 8% (IDEXX) to 33% (Bio-Rad SG) of samples from preclinical goats. Importantly, only IDEXX picked up all samples from clinical bovine spongiform encephalopathy (BSE)-infected goats, whereas the other 2 rapid tests missed 15% (Bio-Rad SG) to 25% (Bio-Rad SAP). These results show that a fraction of preclinical scrapie infections are likely missed by EU surveillance, with sensitivity of detection strongly dependent on the choice of the rapid test. Moreover, a significant proportion of clinical BSE infections are underestimated by using either Bio-Rad test. Assuming that the same sensitivity on preclinical goats would also occur in BSE-infected goats, our data suggest that IDEXX is likely the most sensitive test for detecting preclinical field cases of BSE infection in goats, although with an 8% failure rate. These results raise some concerns about the reliability of current EU surveillance figures on BSE infection in goats.

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confidence: 99%

EU-approved rapid tests might underestimate bovine spongiform encephalopathy infection in goats

et al. 2017

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“…The donor goats were closely monitored for clinical signs of scrapie and subsequently euthanized by venous administration of a commercially available barbiturate solution. Postmortem tissues were fixed in formalin and processed for assessment of PrP Sc accumulation by scrapie IHC (described below) [ 21 , 22 ].…”

Section: Methodsmentioning

confidence: 99%

Low-volume goat milk transmission of classical scrapie to lambs and goat kids

Madsen–Bouterse

Highland²,

Dassanayake³

et al. 2018

PLoS ONE

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The risk of classical scrapie transmission in small ruminants is highest during the neonatal period with the placenta recognized as a significant source of infection. Milk has also been identified as a source of scrapie with sheep-to-sheep transmission occurring after neonatal consumption of as little as 1–2 liters of milk; concurrent mastitis due to small ruminant lentivirus (SRLV) infection may be associated with increased scrapie transmission via milk in sheep. In contrast, goat-to-sheep transmission has been documented only after prolonged consumption of >30 liters of milk. The goal of the current study was to assess transmission of scrapie to goat kids and lambs following low volume, short duration consumption of milk from infected goats. Milk from two does (female goats) with pre-clinical scrapie was fed to four goat kids (≤4.5 L each) and four lambs (~3.7 L each) beginning ~24 hours after birth. Scrapie transmission was detected in three sheep as early as 18 months post inoculation; transmission was also detected in two goats but not until postmortem analyses at 33 months post inoculation. Each milk donor goat also had naturally-acquired infection with SRLV. Different degrees of lymphohistiocytic inflammation and PrPSc accumulation were observed in mammary gland tissues of the donors, which appeared to associate with transmission of scrapie via milk. Thus, similar to the risks of milk transmission of scrapie from sheep, even limited exposure to milk from goats can pose significant risk for scrapie transmission to both goat kids and lambs.

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“…Furthermore, identification of scrapie status of goats can be complicated due to the existence of several polymorphisms in the PRNP coding sequence which can delay the scrapie incubation period and PrP Sc accumulation (Dassanayake et al, 2015;Goldmann et al, 2011;Lacroux et al, 2014). A recent study by us revealed that the polymorphisms in caprine PRNP can also affect the sensitivity of PrP Sc detection in brain samples by anti-prion mAb-based immunoassays such as IHC and Western blot analysis (Madsen-Bouterse et al, 2015). It therefore becomes important to have a rapid, sensitive, PRNP polymorphism-independent as well as antibody-independent detection test to detect scrapie.…”

Section: Discussionmentioning

confidence: 99%

“…Scrapie-uninfected donor goats (n53) were born and housed in a scrapie-free farm at Washington State University with no direct contact with scrapie-infected animals. Of the scrapie-infected donor goats, those bearing WT prion gene (PRNP) haplotypes (n53) were born and raised by scrapie-infected does at the scrapie quarantine farm of the US Department of Agriculture (USDA) Animal Disease Research Unit (Pullman, WA, USA), whereas those heterozygous PRNP with a minor allele (n53) had been referred to us by the USDA Animal and Plant Health Inspection Service (Madsen-Bouterse et al, 2015;O'Rourke et al, 2012). The PRNP genotypes of goats were determined by sequencing the ORF of PRNP as described previously (White et al, 2008).…”

Section: Methodsmentioning

confidence: 99%

“…The prion gene (PRNP) in goats is also highly polymorphic (Vaccari et al, 2009;White et al, 2008). Although goats homozygous for PRNP haplotypes 1 and 2 (WT) are susceptible to scrapie infection, scrapie inoculation studies in goats revealed that PRNP polymorphisms at codons 127,146,154,211 and 222 may provide a prolonged incubation period or possibly resistance to scrapie infection (Acutis et al, 2012;Dassanayake et al, 2015;Lacroux et al, 2014;White et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

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Sensitive and specific detection of classical scrapie prions in the brains of goats by real-time quaking-induced conversion

Dassanayake

Orrú

Hughson

et al. 2016

Journal of General Virology

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Real-time quaking-induced conversion (RT-QuIC) is a rapid, specific and highly sensitive prion seeding activity detection assay that uses recombinant prion protein (rPrP Sen ) to detect subinfectious levels of the abnormal isoforms of the prion protein (PrP Sc ). Although RT-QuIC has been successfully used to detect PrP Sc in various tissues from humans and animals, including sheep, tissues from goats infected with classical scrapie have not yet been tested. Therefore, the aims of the present study were to (1) evaluate whether prion seeding activity could be detected in the brain tissues of goats with scrapie using RT-QuIC, (2) optimize reaction conditions to improve scrapie detection in goats, and (3) compare the performance of RT-QuIC for the detection of PrP Sc with the more commonly used ELISA and Western blot assays. We further optimized RT-QuIC conditions for sensitive and specific detection of goat scrapie seeding activity in brain tissue from clinical animals. When used with 200 mM sodium chloride, both full-length sheep rPrP Sen substrates (PrP genotypes A 136 R 154 Q 171 and V 136 R 154 Q 171 ) provided good discrimination between scrapie-infected and normal goat brain samples at 10 23 dilution within 15 h. Our findings indicate that RT-QuIC was at least 10 000-fold more sensitive than ELISA and Western blot assays for the detection of scrapie seeding activity in goat brain samples. In addition to PRNP WT samples, positive RT-QuIC reactions were also observed with three PRNP polymorphic goat brain samples (G/S127, I/M142 and H/R143) tested. Taken together, these findings demonstrate that RT-QuIC sensitively detects prion seeding activity in classical scrapie-infected goat brain samples.

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PRNP variants in goats reduce sensitivity of detection of PrP^Sc by immunoassay

Cited by 10 publications

References 38 publications

EU-approved rapid tests might underestimate bovine spongiform encephalopathy infection in goats

EU-approved rapid tests might underestimate bovine spongiform encephalopathy infection in goats

Low-volume goat milk transmission of classical scrapie to lambs and goat kids

Sensitive and specific detection of classical scrapie prions in the brains of goats by real-time quaking-induced conversion

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PRNP variants in goats reduce sensitivity of detection of PrPSc by immunoassay

Cited by 10 publications

References 38 publications

EU-approved rapid tests might underestimate bovine spongiform encephalopathy infection in goats

EU-approved rapid tests might underestimate bovine spongiform encephalopathy infection in goats

Low-volume goat milk transmission of classical scrapie to lambs and goat kids

Sensitive and specific detection of classical scrapie prions in the brains of goats by real-time quaking-induced conversion

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PRNP variants in goats reduce sensitivity of detection of PrP^Sc by immunoassay