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
Plasmid Genotype Insert size (bp) Vector Source
pB19-N8
1
5224
pCR-Blunt II-TOPO (Kan)
pCR-Blunt II-TOPO (Kan) pcDNA2.1 (Amp) pcDNA3.1 (Amp)
Zhi et al. 2004
Nguyen et al. 2002 Nguyen et al. 1999 Servant et al. 2002
pA6-cl8
2
4915
pV9 pD91.1
3 3
5028 5035
Evaluation of Plasmids
Transformed each plasmid & maxi preps grown up for each clone
DNA sequence analysis of insert/vector region determined Restriction enzyme digestions of clones
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 used for the validation of assays & material not to be distributed beyond the recipients laboratory
MTA with NIH for pB19-N8, pA6 Hôpital Armand Trousseau permission to supply pV9 & pD91.1