Recognition and partial genome characterization by non-specific DNA amplification and PCR of a new siadenovirus species in a sample originating from Parus major, a great tit

A seemingly novel siadenovirus species was detected by PCR and sequencing in the sample of a great tit (Parus major) found dead in Hungary. Since the genus Siadenovirus has very few known members so far, further study of the virus was intriguing not only from epizootiological but also from taxonomical aspects. The sample, which had been tested in another PCR survey previously. consisted of less than 50 mu 1 of extracted nucleic acid. To ensure sufficient target DNA for an extended study, the viral genome had to be preserved. To this end, the sample was subjected to a novel method of non-specific DNA amplification. Using the amplified DNA as target, different PCR and sequencing strategies were applied with consensus or specific primers for the study of the central genome part of the putative tit adenovirus. The sequence of supposedly one half (13,628 bp) of the genome was determined including eight full genes between the genes of the IVa2 and hexon proteins. The gene content of the viral genome fragment as well as the results of the phylogenetic analyses with different proteins confirmed the discovery of a new species in the genus Siadenovirus. This is the first report on the detection of an adenovirus in great tits. The methods, described in this work, proved suitable for the recovery of nucleic acid samples that contain irreplaceable microbial genomic DNA but are only available in limited quantities. (C) 2009 Elsevier B.V. All rights reserved.