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PP 2.11
Whatman FTA Cards, designed for collection, transport, archiving and isolation of
nucleic acids at room temperature, inactivate human immunodeficiency virus type
1 (HIV-1) and BVDV (model virus for Human Hepatitis C virus).
Penezina O., Neal H.
Whatman Inc., 63 Community Drive, Sanford, ME, 04073, USA.
Objective
To demonstrate that Whatman FTA cards, designed for collection and transport of nucleic
acids at room temperature could inactivate human immunodeficiency virus type 1 (HIV-1)
and BVDV (HCV model) virus.
Methods
HIV-1 (HTLV-IIIB strain) was titrated in-vitro by CEM-A syncytium assay. The HIV-1 stock
solution used in the current study tested positive for identity by negative stain electron
microscopy and p24 analysis and was free of potential bovine and porcine viral
contaminants.
Human blood was spiked with 10% of stock virus solution. An aliquot of the spiked human
blood was immediately tested as T0 sample. Another aliquot was incubated at 22+5 C for
the duration of the FTA Treatment Step (60 minutes) and tested as a Processing Control.
Treatment FTA samples were generated by applying of 125 microliters of spiked blood
onto FTA cards from 4 different lots (3 lots of indicating and 1 lot of non-indicating FTA
cards). At 60 minutes following the initiation of incubation, 6 ml of virus resuspension
media was added to each blood stain card. Serum-free media served as a negative
control for the titration study.
Upon initiation of viral titrations, one aliquot of each test and control sample was diluted in
serum-free medium as appropriate and assayed by the standard virus titration procedure
(in multiple wells for infectious viral particles using CEM-A indicator cells).
The BVDV stock solution used in current study tested positive for identity when
challenged with polyclonal antisera specific for BVDV and was free of potential bovine,
porcine and equine viral contaminants. The titer of the production lot of virus used in the
study was determined by a plaque assay utilizing BT indicator cells.
POSTERS
Application of blood spiked with 10% of stock BVDV virus solution to FTA cards was
identical to one described for HIV-1 study (see above).
Upon initiation of viral titrations, one aliquot of each test and control sample was diluted in
serum-free medium as appropriate and assayed by the standard virus titration procedure
(in multiple wells for infectious viral particles by BVDV plaque assay using BT indicator
cells).
Results
Upon 60 minutes contact of HIV-1 spiked human blood with Whatman FTA cards, the virus
was reduced to non-detectable levels in all FTA card samples resulting in virus Log10
reductions greater than 2.33-2.81 depending on the lot tested (95% confidence interval
was calculated according to the ICH Topic Q5A).
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