Triplex-Loop-Mediated Isothermal Amplification Combined with a Lateral Flow Immunoassay for the Simultaneous Detection of Three Pathogens of Porcine Viral Diarrhea Syndrome in Swine

Porcine epidemic diarrhea virus (PEDV), porcine bocavirus (PBoV), and porcine rotavirus (PoRV) are associated with porcine viral diarrhea. In this study, triplex loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick (LFD) was established for the simultaneous detection o...

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Bibliographic Details
Main Authors: Yi Hong, Biao Ma, Jiali Li, Jiangbing Shuai, Xiaofeng Zhang, Hanyue Xu, Mingzhou Zhang
Format: Article
Language:English
Published: MDPI AG 2023-06-01
Series:Animals
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Online Access:https://www.mdpi.com/2076-2615/13/12/1910
Description
Summary:Porcine epidemic diarrhea virus (PEDV), porcine bocavirus (PBoV), and porcine rotavirus (PoRV) are associated with porcine viral diarrhea. In this study, triplex loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick (LFD) was established for the simultaneous detection of PEDV, PoRV, and PBoV. The PEDV-<i>gp6</i>, PoRV-<i>vp6</i>, and PBoV-<i>vp1</i> genes were selected to design LAMP primers. The amplification could be carried out at 64 °C using a miniature metal bath within 30 min. The triplex LAMP–LFD assay exhibited no cross-reactions with other porcine pathogens. The limits of detection (LODs) of PEDV, PoRV, and PBoV were 2.40 × 10<sup>1</sup> copies/μL, 2.89 × 10<sup>1</sup> copies/μL, and 2.52 × 10<sup>1</sup> copies/μL, respectively. The consistency between rt-qPCR and the triplex LAMP–LFD was over 99% in field samples testing. In general, the triplex LAMP–LFD assay was suitable for the rapid and simultaneous detection of the three viruses in the field.
ISSN:2076-2615