Zika virus inoculation in vivo

Zika virus (strain Zika virus/H.sapiens-tc/BRA/2015/Brazil_SPH2015; genbank accession number KU321639.1) isolated from the plasma of a human in Brazil in 2015 was passaged once on Vero cells and then titrated by Vero cell plaque assay. The inoculum was adjusted to 5.0 log₁₀ PFU (corresponding to 7.8 log₁₀ RNA) in 1 ml of RPMI-1640 medium and injected intravenously in the saphenous vein. Inocula were back-titrated by plaque assay to verify the administered dose. The inoculum tested negative for mycoplasma contamination by deep sequencing (not shown).

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Coffey L.L., Pesavento P.A., Keesler R.I., Singapuri A., Watanabe J., Watanabe R., Yee J., Bliss-Moreau E., Cruzen C., Christe K.L., Reader J.R., von Morgenland W., Gibbons A.M., Allen A.M., Linnen J., Gao K., Delwart E., Simmons G., Stone M., Lanteri M., Bakkour S., Busch M., Morrison J, & Van Rompay K.K. (2017). Zika Virus Tissue and Blood Compartmentalization in Acute Infection of Rhesus Macaques. PLoS ONE, 12(1), e0171148.

Publication 2017

Assay Human Mycoplasma Plaque Plasma Saphenous vein Strain Vero cell Zika virus

Corresponding Organization : University of California, San Francisco

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Protocol cited in 8 other protocols

Variable analysis

independent variables

Zika virus (strain Zika virus/H.sapiens-tc/BRA/2015/Brazil_SPH2015; genbank accession number KU321639.1) isolated from the plasma of a human in Brazil in 2015

dependent variables

Zika virus titer
Zika virus RNA level

control variables

Vero cells (used for virus passage and plaque assay)
RPMI-1640 medium (used for inoculum preparation)
Saphenous vein (used for intravenous injection)
Mycoplasma contamination (tested by deep sequencing)

positive controls

Inocula back-titrated by plaque assay to verify the administered dose

negative controls

Inoculum tested negative for mycoplasma contamination by deep sequencing

Annotations

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