New PCR test for the molecular detection of avian pneumovirus

This study described the design and sensitivities of PCR tests used to detect DNA copies of an A type avian pneumovirus (APV). The objective was selection of PCR primer combinations which would provide the greatest test sensitivity for later testing of field materials. Sensitivities could be determined because of laboratory access to known quantities of purified full length DNA copies of a complete APV genome. Four new nested PCR tests were designed in the fusion (F) protein [2 tests], small hydrophobic (SH) protein and nucleocapsid (N) protein genes and compared to an established test in the attachment (G) protein gene. Sensitivities were found to differ between the tests with the most sensitive being the established G test. When one of the F PCRs was modified to position pyrimidine residues at all primer 3’ termini, its sensitivity exceeded that of the established G test and was chosen for use in future diagnostic testing.