Li Yuqing, Zhao Yanli, Li Chen, Yang Kankan, Li Zhe, Shang Wenbin, Song Xiangjun, Shao Ying, Qi Kezong, Tu Jian
Anhui Province Engineering Laboratory for Animal Food Quality and Bio-safety, College of Animal Science and Technology, Anhui Agricultural University, Hefei, China.
Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, Anhui Agricultural University, Hefei, China.
Front Vet Sci. 2022 Jul 28;9:949172. doi: 10.3389/fvets.2022.949172. eCollection 2022.
Porcine circovirus type 4 (PCV4) is a newly emerging pathogen that was first detected in 2019 and is associated with diverse clinical signs, including respiratory and gastrointestinal distress, dermatitis and various systemic inflammations. It was necessary to develop a sensitive and specific diagnostic method to detect PCV4 in clinical samples, so in this study, a multienzyme isothermal rapid amplification (MIRA) assay was developed for the rapid detection of PCV4 and evaluated for sensitivity, specificity and applicability. It was used to detect the conserved Cap gene of PCV4, operated at 41°C and completed in 20 min. With the screening of MIRA primer-probe combination, it could detect as low as 10 copies of PCV4 DNA per reaction and was highly specific, with no cross-reaction with other pathogens. Further assessment with clinical samples showed that the developed MIRA assay had good correlation with real-time polymerase chain reaction assay for the detection of PCV4. The developed MIRA assay will be a valuable tool for the detection of the novel PCV4 in clinical samples due to its high sensitivity and specificity, simplicity of operation and short testing time.
猪圆环病毒4型（PCV4）是一种新出现的病原体，于2019年首次被检测到，与多种临床症状相关，包括呼吸和胃肠道不适、皮炎以及各种全身性炎症。开发一种灵敏且特异的诊断方法以检测临床样本中的PCV4很有必要，因此在本研究中，开发了一种多酶等温快速扩增（MIRA）检测方法用于快速检测PCV4，并对其敏感性、特异性和适用性进行了评估。该方法用于检测PCV4的保守Cap基因，在41℃下操作，20分钟内完成。通过筛选MIRA引物-探针组合，其每次反应能够检测低至10拷贝的PCV4 DNA，且具有高度特异性，与其他病原体无交叉反应。对临床样本的进一步评估表明，所开发的MIRA检测方法在检测PCV4方面与实时聚合酶链反应检测方法具有良好的相关性。所开发的MIRA检测方法因其高敏感性、特异性、操作简便和检测时间短，将成为临床样本中新型PCV4检测的有价值工具。
