Fig 1.
Schematic of sample collection from hummingbirds (n = 27) with lesions that were visually consistent with pox viral infections.
Both carcasses (n = 19 birds) and field-caught individuals (n = 8 birds) were included in the study.
Fig 2.
Standard curve developed for absolute quantification of viral DNA copies developed through a triplicate test of 10-fold serial dilutions of Avipoxvirus plasmid using the PCR master mix (TaqMan Universal PCR Master Mix, Thermo Fisher Scientific, Carlsbad, California, USA) used for assay development.
Fig 3.
Standard curve developed for absolute quantification of viral DNA copies developed through a triplicate test of 10-fold serial dilutions of Avipoxvirus plasmid using the PCR master mix (Taqman Fast Advanced Master Mix, Thermo Fisher Scientific, Carlsbad, California, USA) used for sample analysis.
Table 1.
Summary of sample types (n = number of samples; percentage) taken from Anna’s Hummingbirds (n = 26) and a Selasphorus spp. where Avipoxvirus was detected by conventional and real-time polymerase chain reaction assay.
Table 2.
Average ± standard deviation (range) of quantification cycle (Cq) values by sample type for samples taken from Anna’s (n = 26) and Selasphorus spp. (n = 1) Hummingbirds and tested via a real-time polymerase chain reaction assay.
Table 3.
Summary of the assessment of the performance of a real-time PCR assay for detecting Avipoxvirus in samples from Anna’s (n = 26) and Selasphorus spp. (n = 1) Hummingbirds.
Confusion matrices, Cohen's kappa, positive agreement (PA), and negative agreement (NA) were used to test for agreement between the real-time PCR and conventional PCR assays.
Fig 4.
Assessment of performance of a real-time PCR assay for detecting Avipoxvirus in samples from Anna’s (n = 26) and Selasphorus spp. (n = 1) Hummingbirds in comparison to true infection status.
Dots represent the sensitivity of the real-time PCR. F1 score statistics and the number of samples for each sample type are reported next to each sample type. Positive predictive values (PPV) for all sample types were calculated to be 1, except for the pox-like lesion swab of the wing (PPV = 0).