Genetic characterization of a mumps virus isolate during passaging in the amniotic cavity of embryonated chicken eggs

The aim of this study was the molecular characterization of a historical mumps isolate (an alleged individual sample). After RNA extraction and cDNA synthesis, selective nested PCR amplification with specific primers, automated DNA sequencing and RFLP analyses were performed. The relative ratios of the detected virus sequences were determined by GeneScan electrophoresis. Phylogenetic tree based on the 316 nucleotide region of the SH gene of the mumps virus was generated by the neighbor-joining method. Results obtained by the described molecular approach show: (a) there are two mumps virus variants, A and B, detected in the fourth passage of wild type virus in the amniotic cavity of embryonated chicken eggs (ECE); (b) variants A and B belong to different genotypes; (c) variants A and B differ in the HN and NP genes which code for amino acid sequences comprising immunogenic epitopes; (d) variant B contains one or more minor variants. We discuss whether the observed differences between the two variants are a consequence of natural heterogeneity or of laboratory contamination in the early passages.