Recovery of divergent avian bornaviruses from cases of proventricular dilatation disease: Identification of a candidate etiologic agent

Kistler, Amy L; Gancz, Ady; Clubb, Susan; Skewes-Cox, Peter; Fischer, Kael; Sorber, Katherine; Chiu, Charles Y; Lublin, Avishai; Mechani, Sara; Farnoushi, Yigal; Greninger, Alexander; Wen, Christopher C; Karlene, Scott B; Ganem, Don; DeRisi, Joseph L

PCR screening 1 ul of the randomly amplified material generated from these RNA samples was used as input template for ABV consensus PCRs as described above. In parallel, as an independent control for input specimen RNA integrity, PCR for glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA was performed on all specimens using designed based on Friedman-Einat et al [30] and Gallu gallus GAPDH sequence: Gg_GAPDHf: 5'-AGTCATCCCT-GAGCTSAAYGG*GAAGC-3' (bp708-733 in Gallus gallus cDNA (NCBI accession NM_204305), * indicates the junction of GAPDH exon 8 and 9 spanned by this primer), Gg_GAPDHr 5'-ACCATCAAGTC-CACAACACGG-3' (Spans bp 1037-1017 in Gallus gallus GAPDH cDNA (NCBI accession NM_204305), maps to GAPDH exon 12). After PCR results were tallied, clinical information on all specimens tested was unmasked. A complete accounting of ABV, GAPDH PCR results, specimen type and clinical status is provided in Additional file 5: Summary of clinical and molecular data for specimens provided in this study.