RESUMO
We report sequencing of four historical cynosurus mottle virus (CnMoV) isolates, originating from different hosts and locations. The CnMoV genome, ranging from 4417 to 4419 nt, encodes five ORFs. It shares 48.1% nucleotide sequence identity with cocksfoot mottle virus and 69.8% with the recently discovered Poaceae Liege sobemovirus. Phylogenetic analysis supports classification within the genus Sobemovirus. Sequenced CnMoV isolates exhibit 96.4-99.9% identity. Nucleotide substitutions leading to amino acid changes showed no host associations. However, amino acid changes in the coat protein appear to be linked to differences in serological properties. Aphid transmission tests confirmed non-transmissibility, consistent with earlier observations for the English isolate.
Assuntos
Genoma Viral , Vírus de RNA , Filogenia , Sequência de Bases , Aminoácidos/genéticaRESUMO
The vacuum integrity of freeze dryers is critical for attaining adequate process control and maintaining confidence in sterility assurance which is key for the manufacture of sterile pharmaceutical products. Although discussions on the topic have been published, there is no industry standard established that is based on empirical data or that has a justifiable scientific rationale. This article provides a review of the scientific literature in the public domain and most importantly, a perspective from 14 Pharmaceutical companies on the leak rate specifications commonly used in industry. Using this information we recommend a best practice for the lyophilizer leak rate test which includes detailing necessary preparation activities following Steam-In-Place (SIP) sterilization, defining a period of stabilization to eliminate pressure and temperature fluctuations and details of the test conditions and the test period. We conclude that for routine manufacturing practice the operational leak rate should not exceed 20 µbar L/s and we provide additional guidance for large volume and older lyophilisation equipment.