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The sera were tested by plaque neutralization (PN) and microneutralization (MN) assays using the reference virus strain A/whooper swan/R65-2/Germany/2006 (H5N1), wasch was directly taken from an infected swan during the outbreak on Ruegen.
The new H5N1 candidate was tested in animals to confirm that it was no longer highly pathogenic (29 ), and vaccine manufacturers are using the reference virus to develop inactivated vaccines that will be evaluated in phase 1 and 2 clinical trials.
Reactive sera were reanalysed by the World Health Organization-Collaborating Centre for Reference and Research on Influenza (WHO-CC), London by MN assay using the reference virus strains A/bar-headed goose/Qinghai/1A/2005 (H5N1) and A/whooper swan/Mongolia/244/2005 (H5N1), which were isolated from infected birds in Asia.
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For the optimization of the CAEV assay, the reference virus strain CAEV Co and two B1 strains from Slovenia were used (Table 2).
The reference virus strain CAEV Co, virus subtype B2 strain and two Slovenian B1 strains were successfully amplified using the CAEV assay.
For the optimization of the MVV assay, the reference virus strain MVV KV1514 and Slovenian SRLV strains of four different subtypes belonging to genotype A were used.
The vaccine strain was the influenza A/Vietnam/1194/NIBRG-14 (H5N1) reference strain prepared by the NIBSC, and is one of the reference viruses indicated as suitable for use in a mock-up vaccine by the European Medicines Agency (EMEA) [14].
In addition, MN assay was performed by GNRCI using reference virus strains A/New Caledonia/20/99 (H1N1) and A/Wisconsin/67/05 (H3N2) to analyse possible presence of cross-reactive antibodies to human influenza A/H1N1 and A/H3N2.
The virus RNA was extracted by using the Biospin Virus RNA Extraction Kit (BioFlux).
B/Florida/04/2006 was also used as a reference virus in case type B viruses were present among the samples tested, a consequence of virus mistyping or mixture.
(6) Reconstructing a complete virus structure (including the capsid and genome) using the virus particle images.
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