Immunoquantitative PCR for Prion Protein Detection in Sporadic Creutzfeldt–Jakob Disease

Abstract Background: The most common human prion disorder is Creutzfeldt–Jakob disease (CJD); it includes sporadic, familial, iatrogenic, and variant subtypes. Diagnostic tests aim at detection with the highest specificity of very small deposits of abnormal prion protein (PrP). Methods: We used immunoquantitative PCR (iqPCR) to detect proteinase K–resistant PrP (PrPRes) in tissue from the middle frontal gyrus of 7 patients with sporadic CJD and 7 non-CJD cases. We compared iqPCR with routine optimized ELISA, Western blotting, and immunohistochemical analyses. Results: The 4 methods showed similar 100% sensitivity and specificity for the diagnosis of CJD. Along with high specificity, however, iqPCR had a threshold for PrPRes detection at least 10-fold lower than that of the classic ELISA. Conclusions: iqPCR is a new method for PrPRes detection that combines 100% specificity with a detection threshold at least 10-fold lower than classic techniques. This method may improve the detection of minute PrPRes deposits in tissues and body fluids and thus be useful for diagnostic and sterilization applications.

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The First Meta-Analysis of the M129V Single-Nucleotide Polymorphism (SNP) of the Prion Protein Gene (PRNP) with Sporadic Creutzfeldt–Jakob Disease

Cells ◽

10.3390/cells10113132 ◽

2021 ◽

Vol 10 (11) ◽

pp. 3132

Author(s):

Yong-Chan Kim ◽

Byung-Hoon Jeong

Keyword(s):

Prion Protein ◽

Protein Gene ◽

Meta Analysis ◽

Prnp Gene ◽

Case Control ◽

Prion Protein Gene ◽

Nucleotide Polymorphism ◽

Single Nucleotide ◽

Jakob Disease ◽

Sporadic Cjd

Prion diseases are fatal, chronic, and incurable neurodegenerative diseases caused by pathogenic forms of prion protein (PrPSc) derived from endogenous forms of prion protein (PrPC). Several case–control and genome-wide association studies have reported that the M129V polymorphism of the human prion protein gene (PRNP) is significantly associated with susceptibility to sporadic Creutzfeldt–Jakob disease (CJD). However, since some case–control studies have not shown these associations, the results remain controversial. We collected data that contain the genotype and allele frequencies of the M129V single-nucleotide polymorphism (SNP) of the PRNP gene and information on ethnic backgrounds from sporadic CJD patients. We performed a meta-analysis by collecting data from eligible studies to evaluate the association between the M129V SNP of the PRNP gene and susceptibility to sporadic CJD. We found a very strong association between the M129V SNP of the PRNP gene and susceptibility to sporadic CJD using a meta-analysis for the first time. We validated the eligibility of existing reports and found severe heterogeneity in some previous studies. We also found that the MM homozygote is a potent risk factor for sporadic CJD compared to the MV heterozygote in the heterozygote comparison model (MM vs. MV, odds ratio = 4.9611, 95% confidence interval: 3.4785; 7.0758, p < 1 × 10−10). To the best of our knowledge, this was the first meta-analysis assessment of the relationship between the M129V SNP of the PRNP gene and susceptibility to sporadic CJD.

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A Practical Approach to Avoiding Iatrogenic Creutzfeldt-Jakob Disease (CJD) from Invasive Instruments

Infection Control and Hospital Epidemiology ◽

10.1017/ice.2015.53 ◽

2015 ◽

Vol 36 (7) ◽

pp. 844-848 ◽

Cited By ~ 16

Author(s):

Paul Brown ◽

Michael Farrell

Keyword(s):

Prion Protein ◽

Diagnostic Tests ◽

Spinal Fluid ◽

Infect Control Hosp ◽

Practical Approach ◽

Cerebellar Disease ◽

Jakob Disease ◽

Contamination Events

Potential Creutzfeldt-Jakob disease instrument-contamination events continue to occur, causing widespread hospital and patient concern. We propose the use of a combination of diagnostic tests (ie, spinal fluid for 14-3-3 protein or nasal brushing for misfolded prion protein) and instrument handling procedures (ie, using a regional set of dedicated instruments), which if applied to all patients admitted with symptoms of either dementia or cerebellar disease, should eliminate the risk of iatrogenic instrument infection.Infect Control Hosp Epidemiol 2015;36(7):844–848

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Use of 14-3-3 in the diagnosis of Creutzfeldt-Jakob disease

Biochemical Society Transactions ◽

10.1042/bst0300382 ◽

2002 ◽

Vol 30 (4) ◽

pp. 382-386 ◽

Cited By ~ 20

Author(s):

A. J. E. Green

Keyword(s):

Cerebrospinal Fluid ◽

Bovine Spongiform Encephalopathy ◽

Diagnostic Tests ◽

Human Diseases ◽

Transmissible Spongiform Encephalopathies ◽

Spongiform Encephalopathy ◽

Spongiform Encephalopathies ◽

Animal Diseases ◽

Jakob Disease ◽

Sporadic Cjd

The transmissible spongiform encephalopathies include human diseases such as Creutzfeldt-Jakob disease (CJD) and kuru as well as animal diseases such as scrapie and bovine spongiform encephalopathy (BSE). The emergence of variant CJD, which is causally related to BSE, has generated much interest in the development of rapid and sensitive diagnostic tests for the pre-mortem diagnosis of CJD. In 1986 two proteins were detected in the cerebrospinal fluid (CSF) of patients with sporadic CJD. These proteins were later demonstrated to be members of the 14-3-3 family, and tests for the detection of CSF 14-3-3 were developed. A number of studies have shown that the detection of CSF 14-3-3 is an accurate test for sporadic CJD, although the results with variant CJD are less promising.

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Prion protein: detection in ‘spiked’ anaerobic sludge and degradation experiments under anaerobic conditions

Water Science & Technology ◽

10.2166/wst.2006.239 ◽

2006 ◽

Vol 53 (8) ◽

pp. 91-98 ◽

Cited By ~ 23

Author(s):

R. Kirchmayr ◽

H.E. Reichl ◽

H. Schildorfer ◽

R. Braun ◽

R.A. Somerville

Keyword(s):

Prion Protein ◽

Screening Method ◽

Protein Detection ◽

Steam Pressure ◽

Proteinase K ◽

Transmissible Spongiform Encephalopathies ◽

Anaerobic Sludge ◽

Spongiform Encephalopathies ◽

Reduction Time ◽

Clinical Phase

The behavior of the transmissible spongiform encephalopathies (TSE) causing agent denominated “prion protein” in anaerobic sludge (biogas reactor) was assessed with incubation tests. A widely applied screening method for BSE in cattle on the basis of the Western blotting protocol was adapted to detect the Proteinase K resistant, scrapie-form prion protein (PrPSC). As PrPSC source homogenized TSE infected brain tissue of animals late in the clinical phase of disease was taken (301V/VM mouse-BSE; bovine BSE and 22A/SV mouse-scrapie). The incubation under mesophilic conditions did not show any significant reduction of the PrPSC titer. Under thermophilic conditions contradictory results were obtained. The reduction time of PrPSC in water was equal to or longer than the PrPSC reduction time in anaerobic sludge. In comparison, with sterilized (121 °C, steam pressure) or poisoned (sodium azide, 1% w/v) sludge used as incubation matrix a much shorter time resulted until no prion protein could be detected.

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Creutzfeldt-Jakob disease associated with a missense mutation at codon 200 of the prion protein gene in Brazil

Dementia & Neuropsychologia ◽

10.1590/s1980-57642008dn10200017 ◽

2007 ◽

Vol 1 (2) ◽

pp. 222-224

Author(s):

Jerusa Smid ◽

Vilma Regina Martins ◽

Michele Christine Landemberger ◽

Daniele Riva ◽

Renato Anghinah ◽

...

Keyword(s):

Cognitive Decline ◽

Missense Mutation ◽

Prion Protein ◽

Clinical Picture ◽

Protein Gene ◽

Clinical Presentation ◽

Prion Protein Gene ◽

Periodic Activity ◽

Jakob Disease ◽

Sporadic Cjd

Abstract Genetic Creutzfeldt-Jakob disease (gCJD) represents less than 15% of CJD cases, and its clinical picture may be either indistinguishable from that of sporadic CJD (sCJD) or be atypical, usually with younger onset and longer duration. We report a case of 59-year old Brazilian man who presented rapidly progressive cognitive decline and cerebellar ataxia. EEG revealed periodic activity. A brother and a cousin of the patient had CJD. A point mutation at codon 200 (E200K) of the prion protein gene (PRNP) was found and death occurred 11 months after onset of symptoms. Autopsy was not performed. The clinical presentation of gCJD associated with E200K, which is the most frequent PRNP mutation, is quite similar to sCDJ. This is the first report of E200K mutation in Brazil, and it is possible that a more systematic search for its occurrence may show it to be relatively frequent in Brazil.

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Frequent Detection of Pituitary-Derived PrPres in Human Prion Diseases

Journal of Neuropathology & Experimental Neurology ◽

10.1093/jnen/nlz075 ◽

2019 ◽

Vol 78 (10) ◽

pp. 922-929

Author(s):

Hiroyuki Honda ◽

Masaki Matsumoto ◽

Masahiro Shijo ◽

Hideomi Hamasaki ◽

Shoko Sadashima ◽

...

Keyword(s):

Growth Hormone ◽

Prion Protein ◽

Prion Diseases ◽

Human Growth ◽

Pituitary Hormones ◽

Cellular Prion Protein ◽

Proteinase K ◽

Molecular Weights ◽

Pituitary Glands ◽

Jakob Disease

Abstract Human prion diseases including sporadic Creutzfeldt-Jakob disease (sCJD), inherited prion diseases, and acquired human prion diseases are lethal neurodegenerative diseases. One of the major sources of iatrogenic Creutzfeldt-Jakob disease was human growth hormone (hGH-iCJD) derived from contaminated cadaveric pituitaries. The incidence of hGH-iCJD has decreased since changing from growth hormone extracted from human cadaveric pituitaries to recombinant pituitary hormones. However, extensive analysis on the localization and detecting of abnormal prion protein in the pituitary gland are limited. In this study, we examined 9 autopsied brains and pituitary glands from 6 patients with prion disease (3 Gerstmann-Sträussler-Scheinker disease, 2 sCJD, and 1 dura mater graft-associated CJD) and 3 individuals with nonprion diseases. Western blot analysis of pituitary samples demonstrated unique glycoforms of normal cellular prion protein with molecular weights of 30–40 kDa, which was higher than the typical 25–35 kDa prion protein in brains. Proteomic analysis also revealed prion protein approximately the molecular weight of 40 kDa in pituitary samples. Moreover, proteinase K-resistant Prion protein was frequently detected in pituitary samples of the prion diseases. Immunohistochemistry for Prion protein revealed mosaic cellular distribution preferentially in growth hormone- or prolactin-producing cells.

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Characterization of Prion Disease Associated with a Two-Octapeptide Repeat Insertion

Viruses ◽

10.3390/v13091794 ◽

2021 ◽

Vol 13 (9) ◽

pp. 1794

Author(s):

Nicholas Brennecke ◽

Ignazio Cali ◽

Tze Mok ◽

Helen Speedy ◽

Laszlo Hosszu ◽

...

Keyword(s):

Prion Protein ◽

Prion Disease ◽

Genetic Variant ◽

Molecular Subtype ◽

Case Series ◽

Low Risk ◽

Proteinase K ◽

Jakob Disease ◽

Insight Into

Genetic prion disease accounts for 10–15% of prion disease. While insertion of four or more octapeptide repeats are clearly pathogenic, smaller repeat insertions have an unclear pathogenicity. The goal of this case series was to provide an insight into the characteristics of the 2-octapeptide repeat genetic variant and to provide insight into the risk for Creutzfeldt–Jakob disease in asymptomatic carriers. 2-octapeptide repeat insertion prion disease cases were collected from the National Prion Disease Pathology Surveillance Center (US), the National Prion Clinic (UK), and the National Creutzfeldt–Jakob Disease Registry (Australia). Three largescale population genetic databases were queried for the 2-octapeptide repeat insertion allele. Eight cases of 2-octapeptide repeat insertion were identified. The cases were indistinguishable from the sporadic Creutzfeldt–Jakob cases of the same molecular subtype. Western blot characterization of the prion protein in the absence of enzymatic digestion with proteinase K revealed that 2-octapeptide repeat insertion and sporadic Creutzfeldt–Jakob disease have distinct prion protein profiles. Interrogation of large-scale population datasets suggested the variant is of very low penetrance. The 2-octapeptide repeat insertion is at most a low-risk genetic variant. Predictive genetic testing for asymptomatic blood relatives is not likely to be justified given the low risk.

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A field on fire: The biochemistry of mad cows

The Biochemist ◽

10.1042/bio02704006 ◽

2005 ◽

Vol 27 (4) ◽

pp. 6-8

Author(s):

David R. Brown

Keyword(s):

Bovine Spongiform Encephalopathy ◽

Prion Protein ◽

Prion Disease ◽

Prion Diseases ◽

Human Diseases ◽

Spongiform Encephalopathy ◽

Related Disease ◽

Jakob Disease ◽

Sporadic Cjd

Prion diseases are neurodegenerative diseases1 that have been linked together because they may potentially have the same cause. These include the diseases scrapie of sheep and BSE (bovine spongiform encephalopathy) of cattle, and also several human diseases that include sporadic CJD (Creutzfeldt-Jakob) disease and a variety of inherited forms. The inherited forms of prion diseases are linked to mutations within the gene for the prion protein. Around 85% of all human cases of prion disease are sporadic CJD, which is a disease affecting people of around 60 years of age. The cause of this disease remains unknown. Unfortunately, the name of this disease causes some confusion, as it is similar to vCJD (variant CJD), a related disease of much younger people.

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