Detection of bat coronavirus using RT-PCR: RNA was extracted from the bat specimens using the MagMAX pathogen RNA/DNA isolation kit (Invitrogen, USA). RT-PCR was performed using Superscript III one-step RT-PCR (Invitrogen, USA) with Platinum High-Fidelity Taq polymerase (Invitrogen, USA) using the published BtCoV-specific primers targeting the conserved region of RNA-dependent RNA polymerase (RdRp) gene21. The amplicon of 440 bp was separated on 1.5 per cent agarose gel and visualized under VersaDoc MP 4000 ultraviolet transilluminator (Bio-Rad, USA).