Please use this identifier to cite or link to this item:
https://hdl.handle.net/1959.11/21248
Title: | Comparison of Commercial Real-Time Reverse Transcription-PCR Assays for Reliable, Early, and Rapid Detection of Heterologous Strains of Porcine Reproductive and Respiratory Syndrome Virus in Experimentally Infected or Noninfected Boars by Use of Different Sample Types | Contributor(s): | Gerber, Priscilla Freitas (author) ; O'Neill, Kevin (author); Owolodun, Olajide (author); Wang, Chong (author); Harmon, Karen (author); Zhang, Jianqiang (author); Halbur, Patrick G (author); Zhou, Lei (author); Meng, Xiang-Jin (author); Opriessnig, Tanja (author) | Publication Date: | 2013 | Open Access: | Yes | DOI: | 10.1128/JCM.02685-12 | Handle Link: | https://hdl.handle.net/1959.11/21248 | Abstract: | The aims of this study were to compare three commercial porcine reproductive and respiratory syndrome virus (PRRSV) realtime reverse transcription-PCR (RT-PCR) assays for detection of genetically diverse PRRSV isolates in serum, semen, blood swabs, and oral fluids collected from experimentally infected boars and to evaluate the effects of sample pooling. Six groups of three boars negative for PRRSV were each inoculated with one of six PRRSV isolates (sharing 55 to 99% nucleotide sequence identity in ORF5). Samples were collected on days-2, 1, 3, 5, 7, 14, and 21 postinoculation (p.i.) and tested by one of three commercially available real-time RT-PCR assays (VetMax from Applied Biosystems, Foster City, CA [abbreviated AB]; VetAlert from Tetracore, Rockville,MD[TC]; and AcuPig from AnDiaTec GmbH, Kornwestheim, Germany [AD]). At day 1 p.i., all assays detected at least one positive sample in each group. The highest detection rates were on days 3 and 5 p.i. Between days 1 and 7 p.i., serum samples had the highest detection rate (90%) with 100% agreement between tests, followed by blood swabs (kappa value of 0.97) and semen (kappa value of 0.80). Oral fluids had the lowest detection rates (AB, 55%; TC, 41%; AD, 46%) and the highest disagreement between kits (kappa value of 0.63). Pools of five samples did not reduce the detection rates if there was one positive sample with a large amount (cycle threshold,<30) of viral RNA in the pool. Serum and blood swab samples had shorter turnaround times for RNA extraction. The AB assay had a 1.6-times-shorter PCR time. In summary, serum and blood swabs had the best performance with highest detection rates and agreement between assays and the shortest turnaround times. | Publication Type: | Journal Article | Source of Publication: | Journal of Clinical Microbiology, 51(2), p. 547-556 | Publisher: | American Society for Microbiology | Place of Publication: | United States of America | ISSN: | 1098-660X 0095-1137 |
Fields of Research (FoR) 2008: | 070712 Veterinary Virology 070703 Veterinary Diagnosis and Diagnostics 070706 Veterinary Medicine |
Fields of Research (FoR) 2020: | 300914 Veterinary virology 300904 Veterinary diagnosis and diagnostics 300907 Veterinary medicine (excl. urology) |
Socio-Economic Objective (SEO) 2008: | 830308 Pigs | Socio-Economic Objective (SEO) 2020: | 100410 Pigs | Peer Reviewed: | Yes | HERDC Category Description: | C1 Refereed Article in a Scholarly Journal |
---|---|
Appears in Collections: | Journal Article |
Files in This Item:
File | Description | Size | Format |
---|
SCOPUSTM
Citations
34
checked on Mar 16, 2024
Page view(s)
926
checked on Mar 9, 2023
Download(s)
2
checked on Mar 9, 2023
Items in Research UNE are protected by copyright, with all rights reserved, unless otherwise indicated.