用于检测 A 组猪轮状病毒的微滴数字 PCR 检测方法的开发。
Development of a droplet digital PCR assay for detection of group A porcine rotavirus.
作者信息
Liu Yangkun, Han Xueying, Zhang Xinru, Liu Jiaxing, Yao Lunguang
机构信息
Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry, School of Life Science and Agricultural Engineering, Nanyang Normal University, Nanyang, China.
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
出版信息
Front Vet Sci. 2023 Mar 6;10:1113537. doi: 10.3389/fvets.2023.1113537. eCollection 2023.
Group A porcine rotavirus (PoRVA) is an important pathogen of acute enteritis in piglets, which has caused severe economic losses in the pig industry worldwide. A convenient, sensitive and specific diagnosis method is an urgent requirement for the surveillance of the PoRVA circulating in clinical samples. In this study, a novel and convenient droplet digital PCR (ddPCR) for the detection of PoRVA was developed using the conserved region of the VP6 gene. The detection limit of ddPCR was 1.81 ± 0.14 copies/rection, ~10 times greater sensitivity than TaqMan real-time quantitative PCR (qPCR). Both ddPCR and qPCR assays exhibited good linearity and repeatability, and the established ddPCR method was highly specific for PoRVA. The results of clinical sample testing showed that the positivity rate of ddPCR (5.6%) was higher than that of qPCR (4.4%). Therefore, the newly developed ddPCR assay could be widely used in clinical diagnosis of PoRVA infections.
A组猪轮状病毒（PoRVA）是仔猪急性肠炎的重要病原体，已在全球养猪业造成严重经济损失。一种便捷、灵敏且特异的诊断方法是监测临床样本中PoRVA流行情况的迫切需求。在本研究中，利用VP6基因的保守区域开发了一种新型便捷的检测PoRVA的液滴数字PCR（ddPCR）方法。ddPCR的检测限为1.81±0.14拷贝/反应，灵敏度比TaqMan实时定量PCR（qPCR）高约10倍。ddPCR和qPCR检测均表现出良好的线性和重复性，且所建立的ddPCR方法对PoRVA具有高度特异性。临床样本检测结果表明，ddPCR的阳性率（5.6%）高于qPCR（4.4%）。因此，新开发的ddPCR检测方法可广泛应用于PoRVA感染的临床诊断。
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