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A cross-sectional assessment of PRRSV nucleic acid detection by RT-qPCR in serum, ear-vein blood swabs, nasal swabs, and oral swabs from weaning-age pigs under field conditions

INTRODUCTION: The porcine reproductive and respiratory syndrome virus (PRRSV) continues to challenge swine production in the US and most parts of the world. Effective PRRSV surveillance in swine herds can be challenging, especially because the virus can persist and sustain a very low prevalence. Alt...

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Autores principales: Osemeke, Onyekachukwu H., Cezar, Guilherme A., Paiva, Rodrigo C., Moraes, Daniel C. A., Machado, Isadora F., Magalhaes, Edison S., Poeta Silva, Ana Paula S., Mil-Homens, Mafalda, Peng, Li, Jayaraman, Swaminathan, Trevisan, Giovani, Silva, Gustavo S., Gauger, Phillip C., Linhares, Daniel C. L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10449646/
https://www.ncbi.nlm.nih.gov/pubmed/37635762
http://dx.doi.org/10.3389/fvets.2023.1200376
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author Osemeke, Onyekachukwu H.
Cezar, Guilherme A.
Paiva, Rodrigo C.
Moraes, Daniel C. A.
Machado, Isadora F.
Magalhaes, Edison S.
Poeta Silva, Ana Paula S.
Mil-Homens, Mafalda
Peng, Li
Jayaraman, Swaminathan
Trevisan, Giovani
Silva, Gustavo S.
Gauger, Phillip C.
Linhares, Daniel C. L.
author_facet Osemeke, Onyekachukwu H.
Cezar, Guilherme A.
Paiva, Rodrigo C.
Moraes, Daniel C. A.
Machado, Isadora F.
Magalhaes, Edison S.
Poeta Silva, Ana Paula S.
Mil-Homens, Mafalda
Peng, Li
Jayaraman, Swaminathan
Trevisan, Giovani
Silva, Gustavo S.
Gauger, Phillip C.
Linhares, Daniel C. L.
author_sort Osemeke, Onyekachukwu H.
collection PubMed
description INTRODUCTION: The porcine reproductive and respiratory syndrome virus (PRRSV) continues to challenge swine production in the US and most parts of the world. Effective PRRSV surveillance in swine herds can be challenging, especially because the virus can persist and sustain a very low prevalence. Although weaning-age pigs are a strategic subpopulation in the surveillance of PRRSV in breeding herds, very few sample types have been validated and characterized for surveillance of this subpopulation. The objectives of this study, therefore, were to compare PRRSV RNA detection rates in serum, oral swabs (OS), nasal swabs (NS), ear-vein blood swabs (ES), and family oral fluids (FOF) obtained from weaning-age pigs and to assess the effect of litter-level pooling on the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) detection of PRRSV RNA. METHODS: Three eligible PRRSV-positive herds in the Midwestern USA were selected for this study. 666 pigs across 55 litters were sampled for serum, NS, ES, OS, and FOF. RT-qPCR tests were done on these samples individually and on the litter-level pools of the swabs. Litter-level pools of each swab sample type were made by combining equal volumes of each swab taken from the pigs within a litter. RESULTS: Ninety-six piglets distributed across 22 litters were positive by PRRSV RT-qPCR on serum, 80 piglets distributed across 15 litters were positive on ES, 80 piglets distributed across 17 litters were positive on OS, and 72 piglets distributed across 14 litters were positive on NS. Cohen's kappa analyses showed near-perfect agreement between all paired ES, OS, NS, and serum comparisons (). The serum RT-qPCR cycle threshold values (Ct) strongly predicted PRRSV detection in swab samples. There was a ≥ 95% probability of PRRSV detection in ES-, OS-, and NS pools when the proportion of positive swab samples was ≥ 23%, ≥ 27%, and ≥ 26%, respectively. DISCUSSION: ES, NS, and OS can be used as surveillance samples for detecting PRRSV RNA by RT-qPCR in weaning-age pigs. The minimum number of piglets to be sampled by serum, ES, OS, and NS to be 95% confident of detecting ≥ 1 infected piglet when PRRSV prevalence is ≥ 10% is 30, 36, 36, and 40, respectively.
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spelling pubmed-104496462023-08-26 A cross-sectional assessment of PRRSV nucleic acid detection by RT-qPCR in serum, ear-vein blood swabs, nasal swabs, and oral swabs from weaning-age pigs under field conditions Osemeke, Onyekachukwu H. Cezar, Guilherme A. Paiva, Rodrigo C. Moraes, Daniel C. A. Machado, Isadora F. Magalhaes, Edison S. Poeta Silva, Ana Paula S. Mil-Homens, Mafalda Peng, Li Jayaraman, Swaminathan Trevisan, Giovani Silva, Gustavo S. Gauger, Phillip C. Linhares, Daniel C. L. Front Vet Sci Veterinary Science INTRODUCTION: The porcine reproductive and respiratory syndrome virus (PRRSV) continues to challenge swine production in the US and most parts of the world. Effective PRRSV surveillance in swine herds can be challenging, especially because the virus can persist and sustain a very low prevalence. Although weaning-age pigs are a strategic subpopulation in the surveillance of PRRSV in breeding herds, very few sample types have been validated and characterized for surveillance of this subpopulation. The objectives of this study, therefore, were to compare PRRSV RNA detection rates in serum, oral swabs (OS), nasal swabs (NS), ear-vein blood swabs (ES), and family oral fluids (FOF) obtained from weaning-age pigs and to assess the effect of litter-level pooling on the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) detection of PRRSV RNA. METHODS: Three eligible PRRSV-positive herds in the Midwestern USA were selected for this study. 666 pigs across 55 litters were sampled for serum, NS, ES, OS, and FOF. RT-qPCR tests were done on these samples individually and on the litter-level pools of the swabs. Litter-level pools of each swab sample type were made by combining equal volumes of each swab taken from the pigs within a litter. RESULTS: Ninety-six piglets distributed across 22 litters were positive by PRRSV RT-qPCR on serum, 80 piglets distributed across 15 litters were positive on ES, 80 piglets distributed across 17 litters were positive on OS, and 72 piglets distributed across 14 litters were positive on NS. Cohen's kappa analyses showed near-perfect agreement between all paired ES, OS, NS, and serum comparisons (). The serum RT-qPCR cycle threshold values (Ct) strongly predicted PRRSV detection in swab samples. There was a ≥ 95% probability of PRRSV detection in ES-, OS-, and NS pools when the proportion of positive swab samples was ≥ 23%, ≥ 27%, and ≥ 26%, respectively. DISCUSSION: ES, NS, and OS can be used as surveillance samples for detecting PRRSV RNA by RT-qPCR in weaning-age pigs. The minimum number of piglets to be sampled by serum, ES, OS, and NS to be 95% confident of detecting ≥ 1 infected piglet when PRRSV prevalence is ≥ 10% is 30, 36, 36, and 40, respectively. Frontiers Media S.A. 2023-08-10 /pmc/articles/PMC10449646/ /pubmed/37635762 http://dx.doi.org/10.3389/fvets.2023.1200376 Text en Copyright © 2023 Osemeke, Cezar, Paiva, Moraes, Machado, Magalhaes, Poeta Silva, Mil-Homens, Peng, Jayaraman, Trevisan, Silva, Gauger and Linhares. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Osemeke, Onyekachukwu H.
Cezar, Guilherme A.
Paiva, Rodrigo C.
Moraes, Daniel C. A.
Machado, Isadora F.
Magalhaes, Edison S.
Poeta Silva, Ana Paula S.
Mil-Homens, Mafalda
Peng, Li
Jayaraman, Swaminathan
Trevisan, Giovani
Silva, Gustavo S.
Gauger, Phillip C.
Linhares, Daniel C. L.
A cross-sectional assessment of PRRSV nucleic acid detection by RT-qPCR in serum, ear-vein blood swabs, nasal swabs, and oral swabs from weaning-age pigs under field conditions
title A cross-sectional assessment of PRRSV nucleic acid detection by RT-qPCR in serum, ear-vein blood swabs, nasal swabs, and oral swabs from weaning-age pigs under field conditions
title_full A cross-sectional assessment of PRRSV nucleic acid detection by RT-qPCR in serum, ear-vein blood swabs, nasal swabs, and oral swabs from weaning-age pigs under field conditions
title_fullStr A cross-sectional assessment of PRRSV nucleic acid detection by RT-qPCR in serum, ear-vein blood swabs, nasal swabs, and oral swabs from weaning-age pigs under field conditions
title_full_unstemmed A cross-sectional assessment of PRRSV nucleic acid detection by RT-qPCR in serum, ear-vein blood swabs, nasal swabs, and oral swabs from weaning-age pigs under field conditions
title_short A cross-sectional assessment of PRRSV nucleic acid detection by RT-qPCR in serum, ear-vein blood swabs, nasal swabs, and oral swabs from weaning-age pigs under field conditions
title_sort cross-sectional assessment of prrsv nucleic acid detection by rt-qpcr in serum, ear-vein blood swabs, nasal swabs, and oral swabs from weaning-age pigs under field conditions
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10449646/
https://www.ncbi.nlm.nih.gov/pubmed/37635762
http://dx.doi.org/10.3389/fvets.2023.1200376
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