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在育肥猪中通过定量实时PCR比较血清池和口腔液样本检测2型猪圆环病毒的情况。

Comparison of serum pools and oral fluid samples for detection of porcine circovirus type 2 by quantitative real-time PCR in finisher pigs.

作者信息

Nielsen Gitte Blach, Nielsen Jens Peter, Haugegaard John, Leth Sanne Christiansen, Larsen Lars E, Kristensen Charlotte Sonne, Pedersen Ken Steen, Stege Helle, Hjulsager Charlotte K, Houe Hans

机构信息

MSD Animal Health Nordic, Havneholmen 25, 1561 Copenhagen V, Denmark.

2Department of Veterinary and Animal Sciences, University of Copenhagen, Grønnegårdsvej 2+8, 1870 Frederiksberg C, Denmark.

出版信息

Porcine Health Manag. 2018 Feb 1;4:2. doi: 10.1186/s40813-018-0079-4. eCollection 2018.

Abstract

BACKGROUND

Porcine circovirus type 2 (PCV2) diagnostics in live pigs often involves pooled serum and/or oral fluid samples for group-level determination of viral load by quantitative real-time polymerase chain reaction (qPCR). The purpose of the study was to compare the PCV2 viral load determined by qPCR of paired samples at the pen level of pools of sera (SP) from 4 to 5 pigs and the collective oral fluid (OF) from around 30 pigs corresponding to one rope put in the same pen. Pigs in pens of 2 finishing herds were sampled by cross-sectional (Herd 1) and cross-sectional with follow-up (Herd 2) study designs. In Herd 1, 50 sample pairs consisting of SP from 4 to 5 pigs and OF from around 23 pigs were collected. In Herd 2, 65 sample pairs consisting of 4 (SP) and around 30 (OF) pigs were collected 4 times at 3-week intervals.

RESULTS

A higher proportion of PCV2-positive pens (86% vs. 80% and 100% vs. 91%) and higher viral loads (mean difference: 2.10 and 1.83 log(10) PCV2 copies per ml) were found in OF versus SP in both herds. The OF cut-off value corresponding to a positive SP (>3 log(10) PCV2 copies per ml) was estimated to 6.5 and 7.36 log(10) PCV2 copies per ml for Herds 1 and 2, respectively. Significant correlations between SP and OF results were found in Herd 1 (rho = 0.69) and the first sampling in Herd 2 (rho = 0.39), but not for the subsequent consecutive 3 samplings in Herd 2.

CONCLUSIONS

The proportion and viral loads of PCV2 positive pens were higher in collective OF (including up to 30 pigs) compared to SP (including 4-5 pigs) of the same pens. Also, OF seemed to detect the PCV2 infection earlier with OF values just below 6.5 (Herd 1) and 7.36 (Herd 2) log(10) being associated with a negative SP for the same pen. Nevertheless, a statistically significant correlation between SP and OF could not be found for all sampling time points, probably due to a high within-pen variation in individual pig viral load becoming very evident in SP of only four or five pigs. Consequently, the results imply that OF is well suited for detecting presence of PCV2 but less so for determining the specific viral load of pigs in a pen.

摘要

背景

对生猪进行猪圆环病毒2型(PCV2)诊断时,通常采用混合血清和/或口腔液体样本,通过定量实时聚合酶链反应(qPCR)在群体水平上测定病毒载量。本研究旨在比较通过qPCR测定的配对样本的PCV2病毒载量,这些配对样本包括来自4至5头猪的血清池(SP)的栏位水平样本,以及来自同一栏位中约30头猪的集体口腔液体(OF)样本。两个育肥猪群栏位中的猪采用横断面研究设计(猪群1)和横断面加随访研究设计(猪群2)进行采样。在猪群1中,收集了50对样本,包括来自4至5头猪的SP和来自约23头猪的OF。在猪群2中,每隔3周收集4次,共收集了65对样本,包括4头猪的(SP)和大约30头猪的(OF)。

结果

在两个猪群中,与SP相比,OF中PCV2阳性栏位的比例更高(分别为86%对80%和100%对91%),病毒载量也更高(平均差异:每毫升2.10和1.83 log(10) PCV2拷贝)。对于猪群1和猪群2,与阳性SP(>每毫升3 log(10) PCV2拷贝)相对应的OF临界值分别估计为每毫升6.5和7.36 log(10) PCV2拷贝。在猪群1(rho = 0.69)和猪群2的首次采样(rho = 0.39)中,SP和OF结果之间存在显著相关性,但在猪群2随后的连续3次采样中未发现相关性。

结论

与同一栏位的SP(包括4 - 5头猪)相比,集体OF(包括多达30头猪)中PCV2阳性栏位的比例和病毒载量更高。此外,OF似乎能更早地检测到PCV2感染,同一栏位中OF值略低于每毫升6.5(猪群1)和7.36(猪群2) log(10)与SP阴性相关。然而,并非在所有采样时间点都能发现SP和OF之间具有统计学意义的相关性,这可能是由于仅4至5头猪的SP中个体猪病毒载量在栏位内变化很大。因此,结果表明OF非常适合检测PCV2的存在,但不太适合确定栏位中猪的具体病毒载量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/42ab/5793352/1505eda5d8c0/40813_2018_79_Fig1_HTML.jpg

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