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首页> 外文期刊>Journal of Veterinary Diagnostic Investigation >Samples sizes required to accurately quantify viral load and histologic lesion severity at the maternal-fetal interface of PRRSV-inoculated pregnant gilts
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Samples sizes required to accurately quantify viral load and histologic lesion severity at the maternal-fetal interface of PRRSV-inoculated pregnant gilts

机译：在PRRSV接种的怀孕Gilts的母体界面下准确地量化病毒载荷和组织学病变严重程度所需的样品尺寸

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Porcine reproductive and respiratory syndrome virus (PRRSV) is transmitted vertically, causing fetal death in late gestation. Spatiotemporal distribution of virus at the maternal-fetal interface (MFI) is variable, and accurate assessment of viral concentration and lesions is thus subject to sampling error. Our objectives were: 1) to assess whether viral load and lesion severity in a single sample of endometrium (END) and placenta (PLC), collected near the base of the umbilical cord (the current standard), are representative of the entire organ; and 2) to compare sampling strategies and evaluate if spatial variation in viral load can be overcome by pooling of like-tissues. Spatially distinct pieces of END and PLC of 24 fetuses from PRRSV-2-infected dams were collected. PRRSV RNA quantified by RT-qPCR was compared in 5 individual pieces per fetus and in respective pools of tissue and extracted RNA. Three distinct pieces of MFI were assessed for histologic severity. Concordance correlation and kappa inter-rater agreement were used to characterize agreement among individual samples and pools. The viral load of individual samples and pools of END had greater concordance to a referent standard than did samples of PLC. Larger pool sizes had greater concordance than smaller pool sizes. Average viral load and lesion severity did not differ by location sampled, and no technical advantages of pooling tissues versus RNA extracts were found. We conclude that multiple pieces of MFI tissues must be evaluated to accurately assess lesion severity and viral load. Three pieces per fetus provided a reasonable balance of cost and logistic feasibility.
机译：猪繁殖与呼吸综合征病毒（PRRSV）垂直传播，导致妊娠晚期胎儿死亡。病毒在母胎界面（MFI）的时空分布是可变的，因此，病毒浓度和病变的准确评估受到采样误差的影响。我们的目标是：1）评估在脐带底部（当前标准）附近采集的单个子宫内膜（END）和胎盘（PLC）样本中的病毒载量和病变严重程度是否代表整个器官；2）比较取样策略，评估病毒载量的空间变异是否可以通过汇集类似组织来克服。收集了来自PRRSV-2感染母鼠的24个胎儿的END和PLC的空间不同片段。通过RT-qPCR定量的PRRSV RNA在每个胎儿的5个单独片段中以及在各自的组织池和提取的RNA中进行比较。对三种不同的MFI进行组织学严重性评估。一致性相关和kappa评分员间一致性被用来描述单个样本和池之间的一致性。与PLC样本相比，单个样本和末端样本池的病毒载量与参考标准的一致性更高。较大的池大小比较小的池大小具有更大的一致性。平均病毒载量和病变严重程度在取样的位置上没有差异，并且没有发现汇集组织与RNA提取物相比的技术优势。我们得出结论，必须对多个MFI组织进行评估，以准确评估病变严重程度和病毒载量。每个胎儿三件可以合理平衡成本和物流可行性。

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