Diagnosis Of Human Prion Diseases Research Articles

Molecular diagnosis of human prion disease

Human prion diseases (PrDs) are transmissible fatal nervous system disorders with public health implications. They are characterized by the presence of a disease-associated form of the physiological cellular prion protein. Development of diagnostic procedures is important to avoid transmission, including through blood products. Methods used for the detection of disease-associated PrP have implications for other neurodegenerative diseases. In this review, the authors discuss recent progress in the understanding of the molecular background of phenotypic variability of human PrDs, and the current concepts of molecular diagnosis. Also, the authors provide a critical summary of the diagnostic methods with regard to the molecular subtypes. In spite of a lack of specific tests to detect disease-associated PrP in body fluids, the constellation of clinical symptoms, detection of protein 14-3-3 in cerebrospinal fluid, electroencephalogram, cranial MRI and prion protein gene examinations, together have increased the specificity and sensitivity of in vivo diagnostics. As new forms of PrDs are reported, continuous evaluation of their incidence and the search for their etiology is crucial. Recent studies, suggesting prion-like properties of certain proteinopathies associated with Parkinson's or Alzheimer's disease, have again brought PrDs to the center of interest as a model of diseases with disordered protein processing.

Rapid screening and confirmatory methods for biochemical diagnosis of human prion disease

Transmissible spongiform encephalopathies (TSEs) are characterised by accumulation of an abnormal isoform of prion protein (PrP sc), mainly in the brain but also in various peripheral tissues. Home-made assays consisting of non-standardised protocols are used currently for laboratory diagnosis of human TSE. The purpose of the present study was to test the ability of two commercial assays, TeSeE™ CJD ELISA and TeSeE™ Western blot, to detect PrPsc in cerebral and lymphoid tissues of TSE patients. Both tests detected a PrPsc-significant signal in the brains of 54 affected patients and not in 51 controls, yielding 100% specificity and 100% sensitivity. Furthermore, three post-mortem spleens and two pre-mortem tonsils from three patients with variant Creutzfeldt–Jakob disease (vCJD) were detected correctly. The expected PrPsc molecular patterns were found in TSE patient brain tissue and in the tonsils and spleens of the three vCJD patients. In conclusion, these rapid and robust in vitro tools were suitable for routine human TSE diagnosis and characterisation. CJD could also be diagnosed during the patient's lifetime by detection of PrPsc in the tonsil. A diagnostic strategy associating TeSeE™ CJD ELISA screening to biochemical confirmation by TeSeE™ Western blot is proposed.

Combined Diffusion Imaging and MR Spectroscopy in the Diagnosis of Human Prion Diseases

The physiopathologic bases underlying the signal intensity changes and reduced diffusibility observed in prion diseases (TSEs) are still poorly understood. We evaluated the interest of MRS combined with DWI both as a diagnostic tool and a way to understand the mechanism underlying signal intensity and ADC changes in this setting. We designed a prospective study of multimodal MR imaging in patients with suspected TSEs. Forty-five patients with a suspicion of TSE and 11 age-matched healthy volunteers were included. The MR imaging protocol included T1, FLAIR, and DWI sequences. MRS was performed on the cerebellum, pulvinar, right lenticular nucleus, and frontal cortex. MR images were assessed visually, and ADC values were calculated. Among the 45 suspected cases, 31 fulfilled the criteria for probable or definite TSEs (19 sCJDs, 3 iCJDs, 2 vCJDs, and 7 genetic TSEs); and 14 were classified as AltDs. High signals in the cortex and/or basal ganglia were observed in 26/31 patients with TSEs on FLAIR and 29/31 patients on DWI. In the basal ganglia, high DWI signals corresponded to a decreased ADC. Metabolic alterations, increased mIns, and decreased NAA were observed in all patients with TSEs. ADC values and metabolic changes were not correlated; this finding suggests that neuronal stress (vacuolization), neuronal loss, and astrogliosis do not alone explain the decrease of ADC. MRS combined with other MR imaging is of interest in the diagnosis of TSE and provides useful information for understanding physiopathologic processes underlying prion diseases.

High-b-Value Diffusion MR Imaging and Basal Nuclei Apparent Diffusion Coefficient Measurements in Variant and Sporadic Creutzfeldt-Jakob Disease

DWI using a standard b-value of 1000 s/mm(2) has emerged as the most sensitive sequence for the diagnosis of CJD. The purpose of this study was to investigate whether DWI at a high b-value (b = 3000 s/mm(2)) and ADC measurements in the basal nuclei improve the diagnosis of vCJD and sCJD compared with visual assessment of DWI at a standard b-value (b = 1000 s/mm(2)). Eight patients with vCJD, 9 patients with sCJD, and 5 healthy volunteers underwent DWI at b = 1000 s/mm(2), and 5 vCJD patients, 4 sCJD patients, and 1 growth hormone-related CJD patient underwent DWI at b = 3000 s/mm(2). Two consultant neuroradiologists performed a visual comparison of the b = 1000 and b = 3000 images. Mean MR SI and ADC values were determined for C, P, and DM thalamus ROIs bilaterally at each b-value. SI ratios for each ROI relative to white matter were calculated. In 9 out of 10 patients, the higher b-value images were more sensitive to SI change, particularly in cortex and thalamus, with higher SI ratios at b = 3000 in the DM thalamus. For sCJD at b = 1000, we found significantly lower ADC values in the C and P compared with controls (mean C ADC = 587.3 +/- 84.7 mm(2)/s in sCJD patients versus 722.7 +/- 16.6 mm(2)/s in controls; P = .007), and at b = 3000, the differences were more pronounced. In comparison, in vCJD at b = 1000, ADC values were elevated in the Pu (mean Pu ADC = 837.6 +/- 33.0 mm/s(2) in vCJD patients versus 748.0 +/- 17.3 mm/s(2) in controls; P < .001) but failed to reach significance at b = 3000. Our results demonstrate that b = 3000 DWI, being more sensitive to slowly diffusing tissue water, is more sensitive to pathology in sCJD than is conventional DWI. High-b-value DWI increases confidence in the radiologic diagnosis of human prion disease.

Rapid diagnosis of human prion disease using streptomycin with tonsil and brain tissues

The use of streptomycin in the pathological prion protein (PrPsc) detection procedures represents a new and attractive way for diagnostic purpose. With this agent, western blot readily detected PrPsc in 263K scrapie hamster and C57Bl/6 wild-type mice challenged with C506M3 scrapie strain. Our aim was to evaluate this new diagnosis procedure in the field of human transmissible spongiform encephalopathies (TSEs). First, we had confirmed the ability of streptomycin to precipitate PrPres from human brain of Creutzfeldt–Jakob disease (CJD) patient. Second, we compared the detection of PrPres with streptomycin against three other protocols using other precipitations. Then we assessed PrPres detection with streptomycin in 98 brain tissue samples from various aetiologies of human TSEs and 52 brain samples from other dementia. Finally, we applied this protocol for tonsils examination of five patients suspected of variant CJD (v-CJD). Sensitivity and specificity obtained with the streptomycin protocol were both 100% on brain tissue. For tonsil tissues, PrPres was clearly identified in the two post-mortem confirmed v-CJD cases, whereas no characteristic three-band pattern was seen in the three confirmed non-v-CJD samples. In this study, streptomycin demonstrated its efficiency to detect PrPres both in the central nervous system and in the lymphoid tissue without practical difficulty and with rapid preparation. Because of its ability to act as a good agent for PrPsc examination in different tissues, recovery of PrPsc in biological fluids using streptomycin should open further perspectives of applications in CJD diagnostics. Streptomycin effects in vivo might thus also be questioned.

Clinical Features and Diagnosis of Human Prion Diseases

Human prion diseases are a group of progressive, fatal encephalopathies united by their similar neuropathological features, their potential transmissibility and a core molecular feature: a conformational change in prion protein. They exist in sporadic/idiopathic, genetic and acquired forms. The clinical features vary according to a number of factors, including cause and the prion protein genotype of the affected individual. Common features include dementia, cerebellar ataxia and involuntary movements. There is no absolute clinical diagnostic test; definitive diagnosis is neuropathological. However, MRI, electroencephalogram, cerebrospinal fluid protein tests, genetic tests and tonsil biopsy, can be very helpful in supporting a clinical diagnosis. The development of a noninvasive diagnostic test (e.g., on blood) would be very helpful in terms of easier, earlier clinical diagnosis, as well as having other applications (such as blood donor screening or population infection prevalence surveys). A relatively new technique (protein misfolding cyclic amplification) holds promise as such a test. Various therapeutic approaches have been suggested, but there is no current effective treatment and there are significant problems in the assessment of any candidate therapies.

In Vivo Detection of Thalamic Gliosis

Increasing evidence supports the usefulness of brain magnetic resonance imaging (MRI) for the diagnosis of human prion diseases. From the neuroradiological point of view, fatal familial insomnia is probably the most challenging to diagnose because brain lesions are mostly confined to the thalamus. To determine whether multisequence MRI of the brain can show thalamic alterations and establish pathoradiologic correlations in a patient with familial fatal insomnia. Radioclinical prospective study. We describe a patient with fatal familial insomnia and normal MRI images. Because the MRI study was performed only 4 days before the patient's death, we were able to compare radiological data with the lesions observed at the neuropathologic level. A 55-year-old man with familial fatal insomnia. Magnetic resonance spectroscopy combined with the measurement of apparent diffusion coefficient of water in different brain areas. The neuroradiological study showed, in the thalamus but not in the other brain regions studied, an increase of apparent diffusion coefficient of water and a metabolic pattern indicating gliosis. These alterations closely correlated with neuropathologic data showing an almost pure gliosis that was restricted to the thalami. Considering fatal familial insomnia as a model of thalamic-restricted gliosis, this case demonstrates that multisequences of magnetic resonance can detect prion-induced gliosis in vivo, as confirmed by a neuropathologic examination performed only a few days after radiological examination.

Sporadic Creutzfeldt–Jakob disease in two adolescents

Background:Sporadic Creutzfeldt–Jakob disease (CJD) is a condition predominantly affecting older age groups, with cases aged less than 45 years rare and an age at onset or death of less than...

Diagnosis of human prion disease

Diagnosis of human prion disease

Diagnosis of human prion disease.

With the discovery of the prion protein (PrP), immunodiagnostic procedures were applied to diagnose Creutzfeldt-Jakob disease (CJD). Before development of the conformation-dependent immunoassay (CDI), all immunoassays for the disease-causing PrP isoform (PrPSc) used limited proteolysis to digest the precursor cellular PrP (PrPC). Because the CDI is the only immunoassay that measures both the protease-resistant and protease-sensitive forms of PrPSc, we used the CDI to diagnose human prion disease. The CDI gave a positive signal for PrPSc in all 10-24 brain regions (100%) examined from 28 CJD patients. A subset of 18 brain regions from 8 patients with sporadic CJD (sCJD) was examined by histology, immunohistochemistry (IHC), and the CDI. Three of the 18 regions (17%) were consistently positive by histology and 4 of 18 (22%) by IHC for the 8 sCJD patients. In contrast, the CDI was positive in all 18 regions (100%) for all 8 sCJD patients. In both gray and white matter, approximately 90% of the total PrPSc was protease-sensitive and, thus, would have been degraded by procedures using proteases to eliminate PrPC. Our findings argue that the CDI should be used to establish or rule out the diagnosis of prion disease when a small number of samples is available as is the case with brain biopsy. Moreover, IHC should not be used as the standard against which all other immunodiagnostic techniques are compared because an immunoassay, such as the CDI, is substantially more sensitive.

P2-187 Hippocampal and amygdalal brain changes in young-old and very-old with Alzheimer's disease: associations with neuropsychological functioning

P2-187 Hippocampal and amygdalal brain changes in young-old and very-old with Alzheimer's disease: associations with neuropsychological functioning

Targeting prion amyloid deposits in vivo.

The diagnosis of prion diseases in humans is challenging due to a lack of specific and sensitive non-invasive tests. Many forms of human prion disease including variant Creutzfeldt-Jakob disease (vCJD), Gerstmann-Sträussler-Scheinker (GSS) syndrome, and 10% of sporadic CJD cases are associated with amyloid deposition. Several positron emission tomography (PET) ligands have recently been developed to directly image beta-amyloid associated with Alzheimer disease. One of them, methoxy-X04, is a fluorescent derivative of Congo red with high binding affinity toward amyloid fibrils and good blood-brain barrier permeability. Using methoxy-X04, we investigated whether amyloid-targeting ligands can be also employed for direct imaging of amyloid deposits associated with some prion diseases. Such a method could potentially become a novel diagnostic approach for these conditions. Studies were performed on MB mice infected with the 87V mouse-adapted scrapie strain. Labeling of PrP amyloid plaques in brains of presymptomatic and symptomatic mice was demonstrated using in vivo transcranial two-photon microscopy after systemic administration of methoxy-X04. During real-time imaging, PrP amyloid deposits could be clearly distinguished 15 min after intravenous administration of methoxy-X04. The ligand showed rapid clearance from brain areas that did not contain amyloid deposits. PrP amyloid deposits could also be detected by direct application of methoxy-X04 on cerebellar sections from GSS patients. These results suggest that methoxy-X04 or similar derivatives could be used as PET imaging agents to improve the diagnosis of human prion diseases associated with amyloid deposition.

Prion diseases in man.

Prion diseases are uncommon fatal neurodegenerative disorders which have gained scientific and public importance as a result of major advances in the understanding of the nature of the causative agent, and the emergence of new forms of these diseases in both animals and man. The transmissible agent in prion diseases is unique and is closely associated with an abnormal isoform of a widely distributed cell-surface glycoprotein, prion protein. The precise mechanisms of conversion to the abnormal isoform are unknown; changes in protein folding are of major importance. The abnormal isoform of the protein accumulates in the central nervous system in all prion diseases, but the processes involved in protein accumulation and the pathogenesis of neuronal dysfunction and cell death are poorly understood. Human prion diseases occur as sporadic, familial, and acquired disorders, the most recently identified of which is new variant Creutzfeldt-Jakob disease, which has been aetiologically linked to exposure to the bovine spongiform encephalopathy agent through the food chain. Surveillance of human prion diseases will be crucial in the assessment of the impact of this new disease in the United Kingdom and elsewhere. Effective surveillance depends on accurate diagnosis, which in turn places a high priority on autopsy in suspected cases; neuropathology is essential for the diagnosis of human prion diseases. Phenotypic variation is prominent in all forms of human prion disease; future classifications of these disorders are likely to incorporate genetic and biochemical data in addition to clinical and pathological parameters.