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商品育肥猪中猪流行性腹泻病毒(PEDV)的检测及抗体反应

Porcine epidemic diarrhea virus (PEDV) detection and antibody response in commercial growing pigs.

作者信息

Bjustrom-Kraft Jordan, Woodard Katie, Giménez-Lirola Luis, Rotolo Marisa, Wang Chong, Sun Yaxuan, Lasley Peter, Zhang Jianqiang, Baum David, Gauger Phillip, Main Rodger, Zimmerman Jeffrey

机构信息

Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, 1850 Christensen Drive, Ames, IA, 50011-1134, USA.

Department of Statistics, College of Liberal Arts and Sciences, Iowa State University, Osborn Drive, Ames, IA, 50011, USA.

出版信息

BMC Vet Res. 2016 Jun 10;12:99. doi: 10.1186/s12917-016-0725-5.

Abstract

BACKGROUND

Longitudinal samples from two production sites were used to (1) describe the pattern of PEDV shedding (rRT-PCR) in individual rectal swabs, pen fecal samples, and pen oral fluids (OF); (2) describe the kinetics of PEDV antibody by ELISA (IgA, IgG) testing of pig serum and pen oral fluid samples; and (3) establish cutoffs and performance estimates for PEDV WV ELISAs (IgA, IgG). Site One was PEDV positive; Site Two was PEDV negative. On Site One, pen samples (feces and oral fluids) and pig samples (rectal swabs and sera) were collected both before and after the population was exposed to PEDV.

RESULTS

On Site Two, pen oral fluid samples and individual pig serum samples were negative for both PEDV antibody and nucleic acid. On Site One, PEDV was detected by rRT-PCR at 6 days post exposure (DPE) in all sample types. The last rRT-PCR positives were detected in rectal swabs and oral fluids on 69 DPE. IgG and IgA were detected in oral fluids and serum samples by 13 DPE. Analysis of the PEDV serum IgG WV ELISA data showed that a sample-to-positive (S/P) cutoff of ≥ 0.80 provided a diagnostic sensitivity of 0.87 (95% CI: 0.82, 0.91) and specificity of 0.99 (95% CI: 0.98, 1.00). Serum IgG results declined slowly over the monitoring period, with 60% of serum samples positive (S/P ≥ 0.80) at the final sampling on 111 DPE. Analysis of the PEDV oral fluid IgA WV ELISA found that a cutoff of S/P ≥ 0.80 provided a diagnostic sensitivity of 1.00 (95% CI: 0.92, 1.00) and a diagnostic specificity of 1.00 (95% CI: 0.99, 1.00). The oral fluid IgA response increased through 96 DPE and began to decline at the last sampling on 111 DPE.

CONCLUSIONS

This study showed that oral fluid-based testing could provide an easy and "animal-friendly" approach to sample collection for nucleic acid and/or antibody-based surveillance of PEDV in swine populations.

摘要

背景

来自两个生产地点的纵向样本用于:(1)描述个体直肠拭子、栏舍粪便样本和栏舍口腔液体(OF)中猪流行性腹泻病毒(PEDV)脱落(逆转录-实时荧光定量聚合酶链反应,rRT-PCR)模式;(2)通过对猪血清和栏舍口腔液体样本进行酶联免疫吸附测定(ELISA,检测IgA、IgG)来描述PEDV抗体的动力学;(3)建立PEDV全病毒ELISA(IgA、IgG)的临界值和性能评估。地点一为PEDV阳性;地点二为PEDV阴性。在地点一,在猪群接触PEDV之前和之后均采集栏舍样本(粪便和口腔液体)以及猪样本(直肠拭子和血清)。

结果

在地点二,栏舍口腔液体样本和个体猪血清样本的PEDV抗体和核酸均为阴性。在地点一,所有样本类型在接触后第6天(DPE)通过rRT-PCR检测到PEDV。在接触后第69天,直肠拭子和口腔液体中检测到最后一批rRT-PCR阳性样本。在接触后第13天,在口腔液体和血清样本中检测到IgG和IgA。对PEDV血清IgG全病毒ELISA数据的分析表明,样本/阳性(S/P)临界值≥0.80时,诊断敏感性为0.87(95%置信区间:0.82,0.91),特异性为0.99(95%置信区间:0.98,1.00)。在监测期内血清IgG结果下降缓慢,在接触后第111天的最后一次采样时,60%的血清样本呈阳性(S/P≥0.80)。对PEDV口腔液体IgA全病毒ELISA的分析发现,S/P临界值≥0.80时,诊断敏感性为1.00(95%置信区间:0.92,1.00),诊断特异性为1.00(95%置信区间:0.99,1.00)。口腔液体IgA反应在接触后第96天前增加,并在接触后第111天的最后一次采样时开始下降。

结论

本研究表明,基于口腔液体的检测可为猪群中PEDV的核酸和/或抗体监测提供一种简便且“对动物友好”的样本采集方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7aba/4902975/0dc3a9b66a14/12917_2016_725_Fig1_HTML.jpg

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