elisa targeting prpcwd  (Bio-Rad)

Journal: Pathogens

Article Title: Evaluating the Diagnostic Efficacy of Using Pooled Samples for Chronic Wasting Disease Testing and Surveillance

doi: 10.3390/pathogens13121133

Figure Lengend Snippet: Figure 1. Results of ELISA (OD of single well, blue triangles) and RT-QuIC (mean time to threshold fluorescence of quadruplicate wells ± one standard error, red circles with bars) testing on pooling thresholds of (A) two deer (e.g., 1:1; n = 6), (B) three deer (e.g., 1:2, n = 10), (C) five deer (e.g., 1:4; n = 6), and (D) 10 deer (e.g., 1:9; n = 17). All pools across assays tested positive, except for P25, which failed to amplify within 24 h on RT-QuIC and is denoted with an asterisk (*).

Article Snippet: Lymph nodes were screened using the commercially available ELISA targeting PrPCWD (TeSeE Purification and Detection Kits, Bio-Rad Laboratories, Inc., Hercules, CA, USA), following the protocols from the manufacturer.

Techniques: Enzyme-linked Immunosorbent Assay, Fluorescence

Journal: Pathogens

Article Title: Evaluating the Diagnostic Efficacy of Using Pooled Samples for Chronic Wasting Disease Testing and Surveillance

doi: 10.3390/pathogens13121133

Figure Lengend Snippet: Figure 2. Results of ELISA (OD of single well, blue triangles) and RT-QuIC (mean time to threshold fluorescence of quadruplicate wells ± one standard error, red circles with bars) for eight positive deer at varied pooling thresholds indicated in parentheses next to the pool ID (A–H). Although Deer ID #16 (D) failed to amplify in one out of four RT-QuIC replicates at a pool size of three (*), it remained otherwise detectable up to a pool size of 10. Overall, OD values across individuals, regardless of pool size, were unpredictable from run to run. In contrast, the times to reach threshold fluorescence increased with pool size and remained fairly repeatable across pool size replicates by individual.

Article Snippet: Lymph nodes were screened using the commercially available ELISA targeting PrPCWD (TeSeE Purification and Detection Kits, Bio-Rad Laboratories, Inc., Hercules, CA, USA), following the protocols from the manufacturer.

Techniques: Enzyme-linked Immunosorbent Assay, Fluorescence

Journal: Pathogens

Article Title: Evaluating the Diagnostic Efficacy of Using Pooled Samples for Chronic Wasting Disease Testing and Surveillance

doi: 10.3390/pathogens13121133

Figure Lengend Snippet: Figure 3. Pool size had a significant fixed effect on assay performance, decreasing OD on ELISA (A) and increasing time, in hours, to reach threshold fluorescence on RT-QuIC (B). The solid lines reflect the fixed effect estimates for pool size from the linear mixed-effects models. Points are colored by positive deer ID and reflect outputs from single wells on ELISA (duplicate mean from pre-screening used for pool size of 1) and the means of quadruplicate wells on RT-QuIC.

Article Snippet: Lymph nodes were screened using the commercially available ELISA targeting PrPCWD (TeSeE Purification and Detection Kits, Bio-Rad Laboratories, Inc., Hercules, CA, USA), following the protocols from the manufacturer.

Techniques: Enzyme-linked Immunosorbent Assay, Fluorescence

Journal: PLOS ONE

Article Title: Are rapid tests and confirmatory western blot used for cattle and small ruminants TSEs reliable tools for the diagnosis of Chronic Wasting Disease in Europe?

doi: 10.1371/journal.pone.0286266

Figure Lengend Snippet: Results obtained from the set 1 of samples by the three ELISA rapid tests at NVI. The O.D. represents the mean of duplicates values obtained from each dilution sample. The values above the cut-off indicate positive sample and those below the cut-off indicate negative sample.

Article Snippet: The first set was analysed with all commercially available ELISA tests TeSeE TM SAP Combi Kit (Bio-Rad), TeSeE TM Sheep/Goats (Bio-Rad), HerdChek BSE-Scrapie Ag test (IDEXX) and HerdChek CWD Ag test (IDEXX).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: PLOS ONE

Article Title: Are rapid tests and confirmatory western blot used for cattle and small ruminants TSEs reliable tools for the diagnosis of Chronic Wasting Disease in Europe?

doi: 10.1371/journal.pone.0286266

Figure Lengend Snippet: Results obtained from the set 2 of samples analyzed at NVI and IRL using three ELISA rapid tests and two western blot methods. The O.D. represents the mean of duplicates /triplicates values obtained from each dilution sample by rapid tests. The values above the cut-off indicate positive sample and those below the cut-off indicate negative sample. Pos and Neg indicate positive and negative results by Western blot analyses.

Article Snippet: The first set was analysed with all commercially available ELISA tests TeSeE TM SAP Combi Kit (Bio-Rad), TeSeE TM Sheep/Goats (Bio-Rad), HerdChek BSE-Scrapie Ag test (IDEXX) and HerdChek CWD Ag test (IDEXX).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Frontiers in Molecular Biosciences

Article Title: Unexpected decrease of full-length prion protein in macaques inoculated with prion-contaminated blood products

doi: 10.3389/fmolb.2023.1164779

Figure Lengend Snippet: Biochemical detection of PrP res in brain samples of experimentally exposed macaques. Frontal, occipital, cerebellar, and medullar samples of the brains from macaques developing the myelopathic syndrome (n = 13, dull green), c-BSE (n = 18, orange), v-CJD (n = 13, red), L-BSE (n = 5, dark green), and MM1 s-CJD (n = 3, purple) or healthy primates (n = 2, blue) were purified using the TeSeE purification kit after serial 10-fold dilutions in healthy brain. After detection, a theoretical optical density of absorbance (450 nm) was calculated by multiplying the O.D. (in the linear part of reading) by the dilution. Results are presented in a 3D plot with theoretical O.D. under a logarithmic scale. Limit of detection is an O.D. = −1 (i.e., O.D. = 0.100).

Article Snippet: Biochemical PrP res analysis: Samples of the frontal cortex, occipital cortex, cerebellum, medulla, and spinal cord were serially 10-fold diluted in corresponding healthy homogenates, and PrP res was purified and detected using the TeSeE ELISA kit (Bio-Rad).

Techniques: Purification

Journal: Frontiers in Molecular Biosciences

Article Title: Unexpected decrease of full-length prion protein in macaques inoculated with prion-contaminated blood products

doi: 10.3389/fmolb.2023.1164779

Figure Lengend Snippet: Biochemical detection of PrP res in spinal cord samples of experimentally exposed macaques. Spinal cord samples from representative macaques developing the myelopathic syndrome (n = 10, dull green), c-BSE (n = 7, orange), and v-CJD (n = 5, red) or healthy primates (n = 2, blue) were tested for the presence of PrP res using the TeSeE kit under the manufacturer’s instructions. Results are presented under a logarithmic scale. The gray zone determines the O.D. below the limit of detection.

Article Snippet: Biochemical PrP res analysis: Samples of the frontal cortex, occipital cortex, cerebellum, medulla, and spinal cord were serially 10-fold diluted in corresponding healthy homogenates, and PrP res was purified and detected using the TeSeE ELISA kit (Bio-Rad).

Techniques:

Journal: Frontiers in Molecular Biosciences

Article Title: Unexpected decrease of full-length prion protein in macaques inoculated with prion-contaminated blood products

doi: 10.3389/fmolb.2023.1164779

Figure Lengend Snippet: Biochemical detection of total PrP in CNS samples of experimentally exposed macaques. The amounts of PrP c in samples of the brain (diamonds) and spinal cord (circles) from macaques developing the myelopathic syndrome (24 cortex samples and 19 spinal cord samples issued from 13 macaques, dull green), c-BSE (12 samples issued from 12 macaques, orange), and v-CJD (33 samples issued from 14 macaques, red) or healthy primates (19 cortex samples and 13 spinal cord samples issued from 7 macaques, blue) were estimated on crude homogenate using the TeSeE kit under the manufacturer’s instructions but without the preliminary step of purification. The six healthy brain samples exhibiting the higher O.D. were serially 10-fold diluted to serve as a reference curve for estimation of the relative amounts of PrP c . The gray zone determines the O.D. below the limit of detection. Significance of Student’s t-test is mentioned between groups.

Article Snippet: Biochemical PrP res analysis: Samples of the frontal cortex, occipital cortex, cerebellum, medulla, and spinal cord were serially 10-fold diluted in corresponding healthy homogenates, and PrP res was purified and detected using the TeSeE ELISA kit (Bio-Rad).

Techniques: Purification

Journal: Frontiers in Molecular Biosciences

Article Title: Unexpected decrease of full-length prion protein in macaques inoculated with prion-contaminated blood products

doi: 10.3389/fmolb.2023.1164779

Figure Lengend Snippet: Effect of denaturation on the biochemical detection of total PrP in CNS samples of experimentally exposed macaques. Crude homogenates of the spinal cord from macaques were mixed with denaturation buffer (C1, TeSeE kit, Bio-Rad) and then heated or not for 5 min at 100°C. The amounts of PrP c in the resulting samples from macaques developing the myelopathic syndrome (19 samples issued from 13 macaques, dull green diamonds), c-BSE (18 samples issued from 9 macaques, orange triangles), and v-CJD (15 samples issued from 5 macaques, red squares) or healthy primates (13 samples issued from 7 macaques, blue circles) were estimated using the TeSeE kit according to the manufacturer’s instructions. The gray zone determines the O.D. below the limit of detection.

Article Snippet: Biochemical PrP res analysis: Samples of the frontal cortex, occipital cortex, cerebellum, medulla, and spinal cord were serially 10-fold diluted in corresponding healthy homogenates, and PrP res was purified and detected using the TeSeE ELISA kit (Bio-Rad).

Techniques:

Journal: Frontiers in Molecular Biosciences

Article Title: Unexpected decrease of full-length prion protein in macaques inoculated with prion-contaminated blood products

doi: 10.3389/fmolb.2023.1164779

Figure Lengend Snippet: Effect of denaturation on the biochemical detection of total PrP in CNS samples of experimentally exposed macaques. Crude homogenates of the spinal cord from macaques were mixed with denaturation buffer (C1, TeSeE kit, Bio-Rad) and then heated or not for 5 min at 100°C. The amounts of PrP c in the resulting samples from macaques developing the myelopathic syndrome (19 samples issued from 13 macaques, dull green diamonds), c-BSE (18 samples issued from 9 macaques, orange triangles), and v-CJD (15 samples issued from 5 macaques, red squares) or healthy primates (13 samples issued from 7 macaques, blue circles) were estimated using the TeSeE kit according to the manufacturer’s instructions. The gray zone determines the O.D. below the limit of detection.

Article Snippet: The detection of PrP by using the ELISA TeSeE Bio-Rad Bovine kit is based on a capture antibody recognizing the octapeptide region and a detection antibody recognizing a C-terminal part located after the hydrophobic core; the absence of detection in myelopathic primates finally shows a deficit of full-length PrP (fl-PrP).

Techniques: