Journal: Pathogens

Article Title: Establishment of a Suitable Diagnostic Workflow to Ensure Sensitive Detection of African Swine Fever Virus Genome in Porcine Semen

doi: 10.3390/pathogens13070537

Figure Lengend Snippet: Semen samples included in the study. The ASF status as true positive (POS) or negative (NEG) was based on manual extraction using the QIAamp Viral RNA Mini kit (QIAGEN) in triplicate, followed by the VetAlert™ PCR as a reference (three independent runs on all replicates). dpi = days post inoculation; NT = not tested.

Article Snippet: Following extraction, all the samples were compared using the VetAlert ASFV DNA assay (Tetracore).

Techniques: Extraction

Journal: Pathogens

Article Title: Establishment of a Suitable Diagnostic Workflow to Ensure Sensitive Detection of African Swine Fever Virus Genome in Porcine Semen

doi: 10.3390/pathogens13070537

Figure Lengend Snippet: Specifications of all ASFV detection (qPCR) protocols and kits used in this study.

Article Snippet: Following extraction, all the samples were compared using the VetAlert ASFV DNA assay (Tetracore).

Techniques: Control

Journal: Pathogens

Article Title: Establishment of a Suitable Diagnostic Workflow to Ensure Sensitive Detection of African Swine Fever Virus Genome in Porcine Semen

doi: 10.3390/pathogens13070537

Figure Lengend Snippet: Comparison of ASFV genome extraction efficiency from boar blood and semen. Evaluation of performance using 18 ASFV-positive blood and 14 positive semen samples. Genome loads were evaluated by qPCR with the VetAlert™ ASFV DNA Test Kit. Boxes represent 25/75 percentiles, including the group median with min. and max. values; each individual is represented by a symbol. Critical EDTA blood (2 dpi) and semen (2–3 dpi) samples are indicated in red. All samples were evaluated in triplicate ( n = 3). N/A = no detection occurred within 45 cycles. Significant differences were assessed by One-Way ANOVA, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Following extraction, all the samples were compared using the VetAlert ASFV DNA assay (Tetracore).

Techniques: Comparison, Extraction

Journal: Pathogens

Article Title: Establishment of a Suitable Diagnostic Workflow to Ensure Sensitive Detection of African Swine Fever Virus Genome in Porcine Semen

doi: 10.3390/pathogens13070537

Figure Lengend Snippet: Analytical sensitivity for boar blood and semen in qPCR. Evaluation of qPCR protocol performance, utilizing 18 ASFV-positive blood and 14 positive semen samples. Boxes represent 25/75 percentiles, including the group median with min. and max. values; each individual is represented by a symbol. Critical EDTA blood (boar #2 and #3 at 2 dpi) and semen (2–3 dpi) samples are indicated in red. Samples were evaluated in triplicate ( n = 3). N/A = no detection occurred within 45 cycles. Significant differences were assessed by One-Way ANOVA, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Following extraction, all the samples were compared using the VetAlert ASFV DNA assay (Tetracore).

Techniques:

Journal: Pathogens

Article Title: Establishment of a Suitable Diagnostic Workflow to Ensure Sensitive Detection of African Swine Fever Virus Genome in Porcine Semen

doi: 10.3390/pathogens13070537

Figure Lengend Snippet: Overview of successfully detected true positive samples among critical semen samples (2–3 dpi). To account for statistical effects, samples were tested in nine replicates in total (three extraction replicates in three PCR runs).

Article Snippet: Following extraction, all the samples were compared using the VetAlert ASFV DNA assay (Tetracore).

Techniques: Extraction

Journal: Pathogens

Article Title: Establishment of a Suitable Diagnostic Workflow to Ensure Sensitive Detection of African Swine Fever Virus Genome in Porcine Semen

doi: 10.3390/pathogens13070537

Figure Lengend Snippet: Comparison of five qPCR assays in ASFV genome detection efficiency from boar blood and semen. Evaluation of qPCR method performance, utilizing 18 ASFV-positive blood and 14 positive semen samples. All qPCR kits were compared to the VetMax™ ASFV Detection Kit. ( A ) Bland–Altman plots. Gray areas represent the lower and upper limits of agreement. Blue points display differences between the VetMax™ ASFV Detection Kit and any other qPCR method tested (WOAH King et al. ). ( B ) Point-by-point result comparison of all samples tested in relation to the VetMax™ ASFV Detection Kit. N/A = no detection occurred within 40–45 cycles. Lines connect results of corresponding samples.

Article Snippet: Following extraction, all the samples were compared using the VetAlert ASFV DNA assay (Tetracore).

Techniques: Comparison

Journal: Pathogens

Article Title: Establishment of a Suitable Diagnostic Workflow to Ensure Sensitive Detection of African Swine Fever Virus Genome in Porcine Semen

doi: 10.3390/pathogens13070537

Figure Lengend Snippet: Evaluation of the detection accuracy of five qPCR kits using semen samples of infected boars. Repeated-measures ANOVA was performed to calculate significant differences in detection performance of the five qPCR kits. Variance between 2 and 3 dpi ( A ), as well as among replicates of each sample ( B ) ( n = 8 with 9 replicates each), was calculated. X-axis labeling corresponds to qPCR kits: A = VetMax™ ASFV Detection Kit, B = VetAlert™ ASFV DNA assay, C = virotype ASFV 2.0 PCR assay, D = RealPCR ASFV DNA Test, E = ASF System 1 (WOAH King et al. ).

Article Snippet: Following extraction, all the samples were compared using the VetAlert ASFV DNA assay (Tetracore).

Techniques: Infection, Labeling

Journal: Frontiers in Pharmacology

Article Title: Detection of Microsatellite Instability in Colorectal Cancer Patients With a Plasma-Based Real-Time PCR Analysis

doi: 10.3389/fphar.2021.758830

Figure Lengend Snippet: Maximum sensitivity evaluation of PNA method. The maximum sensitivity of PNA methods was evaluated using mixed samples of genomic DNA sample obtained from HeLa (MSS) and SNU-1634 (MSI-H) cells. (A) A PNA method was capable of detecting alteration in all five MSI marker genes in sample containing down to 50 pg concentration (A) and (B) down to 1% MSI-H variant.

Article Snippet: We used a peptide nucleotide acid (PNA) probe-mediated real-time PCR-based MSI test, U-TOP MSI Detection Kit Plus (Seasun Biomaterials, Daejeon, Korea), to detect MSI status.

Techniques: Marker, Concentration Assay, Variant Assay

Journal: Frontiers in Pharmacology

Article Title: Detection of Microsatellite Instability in Colorectal Cancer Patients With a Plasma-Based Real-Time PCR Analysis

doi: 10.3389/fphar.2021.758830

Figure Lengend Snippet: Representative MSI test using PNA-mediated melting point analysis (case 20) (A) results from FFPE sample (B) result from plasma sample. Alterations were detected in all five markers in both FFPF and plasma sample.

Article Snippet: We used a peptide nucleotide acid (PNA) probe-mediated real-time PCR-based MSI test, U-TOP MSI Detection Kit Plus (Seasun Biomaterials, Daejeon, Korea), to detect MSI status.

Techniques: