猪肠冠状病毒(SECoV),具有高杀伤力和传染性,是导致仔猪致命和水样腹泻并在全球传播的主要病原体。此外,这些SECoV可以引起类似的临床表现,并且经常共同感染,需要适合快速的准确测定,在原地,和差分检测。这里,我们开发了一种基于多重荧光的侧向流免疫测定法(mFB-LFIA),用于检测三种SECoV,包括猪三角洲冠状病毒(PDCoV),传染性胃肠炎病毒(TGEV),猪流行性腹泻病毒(PEDV),在猪粪便样本中。由于滤波垫的设计和合理的优化,mFB-LFIA在15分钟内同时检测三个SECoV,并提高了条带对粪便样品的耐受性。检测PDCoV的检测限(LoD),TGEV,和PEDV分别为2.1×104TCID50mL-1、3.4×102TCID50mL-1和3.6×102TCID50mL-1。此外,该方法已成功应用于PDCoV的检测,TGEV,和PEDV在猪粪便中具有较高的准确性。与金标准核酸检测相比,拟议测定的总符合率超过90%。此外,mFB-LFIA表现出优异的稳定性和可重复性。拟议的mFB-LFIA允许快速,在原地,更经济有效的同时检测PDCoV,TGEV,和PEDV与核酸检测的比较。据我们所知,这是第一份描述能够检测PDCoV的多重护理点测定的报告,TGEV,和猪粪便样本中的PEDV。我们相信我们的方法在猪场有很大的应用潜力。
Swine Enteric Coronaviruses (SECoVs), with high lethality and infectiousness, are the main pathogens causing fatal and watery diarrhea in piglets and spreading globally. Moreover, these SECoVs can cause similar clinical manifestations and are often co-infected, requiring an accurate assay suitable for rapid, in situ, and differential detection. Here, we developed a multiplexed fluorescent-based lateral flow immunoassay (mFB-LFIA) for the detection of three SECoVs, including porcine delta coronaviruses (PDCoV), transmissible gastroenteritis virus (TGEV), and porcine epidemic diarrhea virus (PEDV), in swine fecal samples. Thanks to the filter pad design and reasonable optimization, the mFB-LFIA was achieved within 15 min for three SECoVs detection simultaneously and improved the tolerance of the strips for feces samples. The limit of detection (LoD) of detecting PDCoV, TGEV, and PEDV were 2.1 × 104 TCID50 mL-1, 3.4 × 102 TCID50 mL-1, and 3.6 × 102 TCID50 mL-1, respectively. Additionally, the proposed assay was successfully applied to the detection of PDCoV, TGEV, and PEDV in swine feces with high accuracy. Compared with the gold standard nucleic acid testing, the total coincidence rate of the proposed assay was more than 90 %. Moreover, the mFB-LFIA performed excellent stability and repeatability. The proposed mFB-LFIA allows for rapid, in situ, more cost-effective and simultaneous detection of PDCoV, TGEV, and PEDV compared with nucleic acid testing. To the best of our knowledge, this is the first report to describe a multiplexed point-of-care assay capable of detecting PDCoV, TGEV, and PEDV in swine fecal samples. We believe our approach has a great potential for application to pig farm.