A collection of Neofabraea isolates from pear fruit grown in Oregon, Washington, and California was screened with species-specific primers in a multiplex polymerase chain reaction (PCR). Neofabraea alba was identified most frequently in samples from Oregon and California, whereas N. perennans was found most frequently in samples from Washington. N. alba also was identified from tissue of small cankers and pruning stubs on pear trees using PCR. Bull's eye rot pathogens were isolated from fruit of nine different European pear cultivars, Asian pear, and quince. Overall, N. alba was the most prevalent species in 2001 whereas N. perennans was more prevalent in 2002. An undescribed species of Neofabraea was identified in samples from Medford, OR that corresponds to a species previously found by others using molecular methods. This information increases the known geographic distribution of this undescribed species.