Evaluation of Cloned DNA for B19 Genotypes Dr Jacqueline Fryer, Division of Virology NIBSC.

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Presentation transcript:

Evaluation of Cloned DNA for B19 Genotypes Dr Jacqueline Fryer, Division of Virology NIBSC

Genetic Diversity of Parvovirus B19: Analysis of the NS1/VP1 Region A6 Servant et al., J. Virol. 2002

Plasmid Information PlasmidGenotype Insert size (bp)VectorSource pB19-N pCR-Blunt II-TOPO (Kan) Zhi et al pA6-cl pCR-Blunt II-TOPO (Kan) Nguyen et al pV935028pcDNA2.1 (Amp) Nguyen et al pD pcDNA3.1 (Amp) Servant et al. 2002

Evaluation of Plasmids Transformed each plasmid & maxi preps grown up for each clone Transformed each plasmid & maxi preps grown up for each clone DNA sequence analysis of insert/vector region determined DNA sequence analysis of insert/vector region determined Restriction enzyme digestions of clones Restriction enzyme digestions of clones Analysis by consensus real-time PCR assay Analysis by consensus real-time PCR assay

Analysis of Plasmid DNAs by Consensus Assay for Parvovirus B19

Distribution and Use of Clones to OMCL Network and Manufacturers To be distributed through the EDQM PTS for parvovirus B19 DNA To be distributed through the EDQM PTS for parvovirus B19 DNA To be used for the validation of assays & material not to be distributed beyond the recipients laboratory To be used for the validation of assays & material not to be distributed beyond the recipients laboratory MTA with NIH for pB19-N8, pA6 MTA with NIH for pB19-N8, pA6 Hôpital Armand Trousseau permission to supply pV9 & pD91.1 Hôpital Armand Trousseau permission to supply pV9 & pD91.1