HCoV-NL63 was first isolated from babies suffering of pneumonia and bronchiolitis in 2004. HCoV-NL63 has been found worldwide with up to 9.3% detection rate in hospitalized respiratory tract samples. In 2010, a bat coronavirus termed ARCoV.2(Appalachian Ridge CoV) detected in North American tricolored bat (Perimyotis subflavus) in the US showed close relationship with HCoV-NL63. The MRCA for HCoV-NL63 and ARCoV.2 was predicted to have existed 563 to 822 years ago. Further analysis indicated that HCoV-NL63 can replicate in cell lines derived from the lungs of tricolored bats. These results suggest that prototypes of HCoV-NL63 may also exist in bats and there may also be a bat origin of this human coronavirus.