Song Wenbo, Feng Yixue, Zhang Jiali, Kong Danni, Fan Jie, Zhao Mengfei, Hua Lin, Xiang Jinmei, Tang Xibiao, Xiao Shaobo, Peng Zhong, Wu Bin
National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, 430070, Wuhan, China.
Hubei Hongshan Laboratory, 430070, Wuhan, China.
Porcine Health Manag. 2024 Mar 5;10(1):12. doi: 10.1186/s40813-024-00364-y.
Diarrheal diseases caused by viral agents have led to a great morbidity, mortality, and economic loss in global pig industry. Porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine deltacoronavirus (PDCoV), and group A porcine rotavirus (RVA) are main causative agents of swine viral diarrhea with similar clinical signs on Chinese farms and their co-infection is also common. However, it is still lack of a convenient method to detect these four agents.
A TaqMan multiplex qPCR method was developed to detect PEDV, TGEV, PDCoV, and RVA, simultaneously. This method was then applied to investigate 7,342 swine fecal samples or rectal swabs, as well as 1,246 swine intestinal samples collected from 2075 farms in China in 2022.
Minimum detection limits of this method were 3 copies/µL for PEDV, 4 copies/µL for TGEV, 8 copies/µL for RVA, and 8 copies/µL for PDCoV, suggesting a good sensitivity. No signals were observed by using this method detecting other viral agents commonly prevalent in pigs, which is suggestive of a good specificity. Application of this method on investigating clinical samples demonstrated a relatively high positive rate for PEDV (22.21%, 1907/8588) and RVA (44.00%, 3779/8588). In addition, co-infection between PEDV and RVA was observed on 360 investigated farms, accounting for 17.35% (360/2075) of the farms where co-infection events were screened.
A TaqMan multiplex qPCR method targeting PEDV, TGEV, PDCoV, and RVA was developed in this study. This method demonstrated a good specificity and sensitivity on investigating these four common viruses responsible for viral diarrhea on Chinese pig farms, which represents a convenient method for the monitoring and differential diagnosis of swine viral diarrhea.
由病毒引起的腹泻疾病给全球养猪业带来了巨大的发病率、死亡率和经济损失。猪流行性腹泻病毒（PEDV）、传染性胃肠炎病毒（TGEV）、猪三角洲冠状病毒（PDCoV）和A组猪轮状病毒（RVA）是中国猪场猪病毒性腹泻的主要病原体，它们具有相似的临床症状，且共同感染也很常见。然而，目前仍缺乏一种便捷的方法来检测这四种病原体。
建立了一种TaqMan多重荧光定量PCR方法，用于同时检测PEDV、TGEV、PDCoV和RVA。该方法随后应用于检测2022年从中国2075个猪场采集的7342份猪粪便样本或直肠拭子以及1246份猪肠道样本。
该方法对PEDV的最低检测限为3拷贝/μL，对TGEV为4拷贝/μL，对RVA为8拷贝/μL，对PDCoV为8拷贝/μL，表明具有良好的敏感性。使用该方法检测猪中常见的其他病毒病原体未观察到信号，提示具有良好的特异性。将该方法应用于临床样本检测显示，PEDV的阳性率相对较高（22.21%，1907/8588），RVA的阳性率为44.00%（3779/8588）。此外，在360个被调查猪场中观察到PEDV和RVA的共同感染，占筛查到共同感染事件猪场的17.35%（360/2075）。
本研究建立了一种针对PEDV、TGEV、PDCoV和RVA的TaqMan多重荧光定量PCR方法。该方法在检测中国猪场中这四种引起病毒性腹泻的常见病毒时表现出良好的特异性和敏感性，是一种用于猪病毒性腹泻监测和鉴别诊断的便捷方法。
