Han Shanshan, Qin Tao, Chen Taichun, Xing Xiaocheng, Zhang Qiang
Enzyme Engineering Research Center of Shaanxi Province, Xi'an 710600, China.
Bio-Agriculture Institute of Shaanxi Province, Shaanxi Academy of Sciences, Xi'an 710043, China.
Vet Anim Sci. 2025 Aug 27;30:100503. doi: 10.1016/j.vas.2025.100503. eCollection 2025 Dec.
Porcine reproductive and respiratory syndrome virus (PRRSV) is an RNA virus that induces reproductive disorders in sows and respiratory diseases in growing pigs. Recently, the NADC34-like strain of PRRSV has become more prevalent, with outbreaks occurring across pig farms in China. However, a reliable diagnostic method for the clinical detection of this strain has been absent. This study developed a TaqMan probe-based dual real-time quantitative PCR assay targeting the M and GP5 genes to specifically identify the NADC34-like PRRSV strain. The assay exhibited high specificity, detecting exclusively the NADC34-like strain without cross-reactivity with other PRRSV strains. The detection limits for pMD-M and pMD-GP5 plasmids were 2.67 × 10² and 1.35 × 10¹ copies/μL, respectively, indicating high assay sensitivity. The assay also demonstrated excellent reproducibility, with coefficient of variation (CV) values for both recombinant plasmids below 2 %. Among 251 clinical samples, 27 tested positive for NADC34-like PRRSV. This study establishes an accurate, sensitive, and reliable TaqMan dual real-time PCR assay for detecting NADC34-like PRRSV, offering a valuable tool for clinical diagnostics and outbreak management in pig farms.
猪繁殖与呼吸综合征病毒（PRRSV）是一种RNA病毒，可导致母猪繁殖障碍和生长猪呼吸道疾病。最近，PRRSV的NADC34样毒株变得更加普遍，在中国的养猪场中爆发。然而，一直缺乏用于临床检测该毒株的可靠诊断方法。本研究开发了一种基于TaqMan探针的双重实时定量PCR检测方法，靶向M基因和GP5基因，以特异性鉴定NADC34样PRRSV毒株。该检测方法具有高特异性，仅能检测到NADC34样毒株，与其他PRRSV毒株无交叉反应。pMD-M和pMD-GP5质粒的检测限分别为2.67×10²和1.35×10¹拷贝/μL，表明该检测方法具有高灵敏度。该检测方法还具有出色的重复性，两种重组质粒的变异系数（CV）值均低于2%。在251份临床样本中，27份检测出NADC34样PRRSV呈阳性。本研究建立了一种准确、灵敏且可靠的用于检测NADC34样PRRSV的TaqMan双重实时PCR检测方法，为猪场的临床诊断和疫情管理提供了有价值工具。
