A simple and rapid identification method for newly emerged porcine Deltacoronavirus with loop-mediated isothermal amplification

Zhang, Fanfan; Ye, Yu; Song, Deping; Guo, Nannan; Peng, Qi; Li, Anqi; Zhou, Xingrong; Chen, Yanjun; Zhang, Min; Huang, Dongyan; Tang, Yuxin

Currently, available methods for detection of PDCoVs include conventional reverse transcription-polymerase chain reaction (RT-PCR), nested RT-PCR, real-time RT-PCR, and ELISA [6, [10] [11] [12] [13] . However, these techniques have some shortcomings, such as higher requirements for equipment, high cost, extended detection period, and/or low sensitivity [14] . Reverse transcription loopmediated isothermal amplification (RT-LAMP) provides a potential effective tool for rapid and accurate identification of viral pathogens, which amplifies nucleic acids under isothermal conditions with high sensitivity and specificity. This novel gene detection technique is costeffective and time-saving, and only requires a constant temperature water bath. RT-LAMP has been widely used in clinical diagnosis for detection of the presence of several important viral pathogens, including PEDV, TGEV, classic swine fever virus (CSFV), and H10N8 subtype of influenza A virus [14] [15] [16] [17] . The improved LAMP assays, such as visual detection of amplified products by adding SYBR Green I or hydroxynaphthol blue (HNB), have made it easier to apply in primary clinical settings or for field use. In the present work, we developed and evaluated a specific and sensitive RT-LAMP assay for visual detection of PDCoV, which might be a good tool for the diagnosis of PDCoV in field samples.