ArticleJournal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc2018
Effects of sample handling on the detection of porcine reproductive and respiratory syndrome virus in oral fluids by reverse-transcription real-time PCR.
Article in Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 12 citations in OpenAlex.
- Full Factorial Comparison of the Diagnostic Performance of Three Nucleic Acid Extraction Kits and Three PRRSV RT-qPCR Assays Using Swine Oral Fluids of Known Status.Microorganisms · 2026Article
- Animal Welfare-Friendly Noninvasive Oral Fluid in Swine Diagnostics: Current Applications in Disease Monitoring, Key Challenges and Future Directions.Veterinary medicine international · 2026Review
- Effect of temperature, relative humidity, and time on the detection of swine RNA viruses (PRRSV, PEDV, IAV) inoculated onto filter papers.Frontiers in cellular and infection microbiology · 2026Article
- Effect of Pooling and Freeze-Thaw Processes on the PRRSV RNA Detection in TOSc Samples.Veterinary sciences · 2025Article
- Effect of Time and Temperature on the Detection of PRRSV RNA and Endogenous Internal Sample Control in Porcine Tongue Fluids.Veterinary sciences · 2025Article
- Effect of stabilizers on the detection of swine influenza A virus (swIAV) in spiked oral fluids over time.Porcine health management · 2024Article
- Critical evaluation of strategies to achieve direct real-time PCR detection of swine pathogens in oral fluids.Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2023Article
- Best practices for performance of real-time PCR assays in veterinary diagnostic laboratories.Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2020Article
- Guidelines for oral fluid-based surveillance of viral pathogens in swine.Porcine health management · 2020Review
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Authors and funding
10 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We evaluated effects of handling procedures on detection of porcine reproductive and respiratory syndrome virus (PRRSV) in oral fluids (OFs) by reverse-transcription real-time PCR (RT-rtPCR). The experiments were conducted using a composite sample of PRRSV-positive OF collected from 5-wk-old pigs vaccinated 15 d earlier with a modified-live PRRSV vaccine. Five pre-extraction sample-handling steps and all combinations thereof were evaluated: 1) thaw temperature (4°C or 25°C); 2) sample diluent (1:1 dilution with nuclease-free water or guanidinium thiocyanate-phenol); 3a) sonication of the sample (yes or no); 3b) temperature (4°C or 25°C) at which step 3a was conducted; and 4) temperature at which the sample was maintained after step 3b and until RNA extraction was initiated (4°C or 25°C). All combinations of the 5 sample-handling steps (i.e., 32 unique treatments) were tested in a completely randomized factorial design with 4 replicates and 1 negative control for each treatment. The entire experiment was repeated on 5 separate days to produce a total of 800 PRRSV RT-rtPCR results. Binary (positive or negative) data were analyzed by logistic regression and results (Ct) were analyzed using a generalized linear model. Overall, 1 false-positive result was observed among 160 negative controls (99.4% specificity), and 85 false-negative results were observed among the 640 known-positive samples (86.7% sensitivity). The most significant factor affecting test outcome was thaw temperature (4°C or 25°C); samples thawed at 4°C had higher positivity rate (94% vs. 80%, p < 0.0001) and lower Ct (36.2 vs. 37.5, p < 0.0001).
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