A filter pad design-based multiplexed lateral flow immunoassay for rapid simultaneous detection of PDCoV, TGEV, and PEDV in swine feces

• Published in: Talanta (Oxford) 2024-12, Vol.280, p.126712, Article 126712
• Main Authors: Wu, Lei, Liang, Jiajie, Teng, Peijun, Du, Yifan, He, Yong, Liao, Shusen, Wang, Juan, Zhang, Xiaoli, Wang, Zhaoguang, Zeng, Tengyue, Wang, Yaqi, Zou, Siyi, Lu, Cheng, Jia, Aiqing, Song, Qifang, Huang, Boyan, Fang, Liurong, Cheng, Wenli, Tang, Yong
• Format: Article
• Language: English
• Subjects: Animals; Deltacoronavirus - isolation & purification; Feces - chemistry; Feces - virology; Highly pathogenic; Immunoassay - methods; Lateral flow immunoassay; Limit of Detection; Porcine epidemic diarrhea virus - isolation & purification; Simultaneous detection; Swine; Swine enteric coronaviruses; Swine feces; Transmissible gastroenteritis virus - isolation & purification
• ISSN: 0039-9140; 1873-3573; 1873-3573
• DOI: 10.1016/j.talanta.2024.126712

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. [Display omitted] •A multiplexed LFIA for the detection of SECoVs in swine feces was first reported.•A filter pad design for multiplexed LFIA to remove the interference of the matrix.•A novel EuNPs-based LFIA was developed for the detection of PDCoV, TGEV and PEDV.