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在田间条件下,对断奶仔猪血清、耳静脉血拭子、鼻拭子和口腔拭子进行RT-qPCR检测猪繁殖与呼吸综合征病毒(PRRSV)核酸的横断面评估。

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.

作者信息

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

机构信息

Fieldepi, Iowa State University College of Veterinary Medicine, Ames, IA, United States.

Veterinary Diagnostic and Production Animal Medicine Department of the College of Veterinary Medicine, Iowa State University, Ames, IA, United States.

出版信息

Front Vet Sci. 2023 Aug 10;10:1200376. doi: 10.3389/fvets.2023.1200376. eCollection 2023.

Abstract

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.

摘要

引言

猪繁殖与呼吸综合征病毒(PRRSV)持续对美国及世界大部分地区的养猪业构成挑战。对猪群进行有效的PRRSV监测具有挑战性,特别是因为该病毒能够持续存在且流行率极低。虽然断奶仔猪是繁殖猪群中PRRSV监测的一个关键亚群,但用于该亚群监测的样本类型经过验证和特征描述的却很少。因此,本研究的目的是比较从断奶仔猪采集的血清、口腔拭子(OS)、鼻拭子(NS)、耳静脉血拭子(ES)和家庭口腔液(FOF)中PRRSV RNA的检测率,并评估窝水平混合对PRRSV RNA逆转录定量聚合酶链反应(RT-qPCR)检测的影响。

方法

在美国中西部选择了三个符合条件的PRRSV阳性猪群进行本研究。对55窝的666头猪采集血清、NS、ES、OS和FOF样本。对这些样本分别以及对拭子的窝水平混合样本进行RT-qPCR检测。每种拭子样本类型的窝水平混合样本是通过将从一窝内的猪采集的等量每种拭子混合而成。

结果

通过PRRSV RT-qPCR检测,血清检测呈阳性的有分布在22窝的96头仔猪,ES检测呈阳性的有分布在15窝的80头仔猪,OS检测呈阳性的有分布在17窝的80头仔猪,NS检测呈阳性的有分布在14窝的72头仔猪。科恩kappa分析显示,所有配对的ES、OS、NS和血清比较之间几乎完全一致()。血清RT-qPCR循环阈值(Ct)强烈预测拭子样本中PRRSV的检测。当阳性拭子样本比例分别≥23%、≥27%和≥26%时,ES、OS和NS混合样本中PRRSV检测的概率≥95%。

讨论

ES、NS和OS可作为通过RT-qPCR检测断奶仔猪PRRSV RNA的监测样本。当PRRSV流行率≥10%时,为有95%的把握检测到≥1头感染仔猪,血清、ES、OS和NS分别需要采样的仔猪最小数量为30头、36头、36头和40头。

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