TABLE 2.

Overview of modified qVOA assays.

Assay Name	Modification/Format	Readout	Reference(s)
QVOA2	PHA+ sero-negative γ-irradiated PBMC stimulation of resting CD4+ T cells; MOLT4/CCR5 or SupTl-CCR5 as targets for virus expansion	PolyA RNA qPCR or p24 ELISA	Laird et al., 2013; Fun et al., 2017
TZA	CD3/CD28 stimulated CD4+ T cells are co-cultured with TZM-bl indicator cells	B-gal activity by chemiluminescence	Sanyal et al., 2017
dQVOA	Resting CD4+ T Cells are differentiated to effector memory phenotype before activating with PHA + seronegative γ-irradiated PBMC; no targets added	p24 ELISA	Wonderlich et al., 2019
mQVOA	Immobilized CD3/CD28 simulation of CD4+ T cells; MOLT4/CCR5 targets; cell free RNA in supernatant captured by magnetic beads	gag or pol RNA qPCR	Massanella et al., 2018; Rosenbloom et al., 2019
Murine VOA (mVOA)	CD4+ T cells or PBMC from aviremic donors engrafted into immunodeficient NSG mice, followed by administration of anti- CD3/CD28 and anti-CD8 antibodies	Plasma HIV RNA detection (gag primers) by qPCR	Metcalf Pate et al., 2015
Humanized mouse VOA (hmVOA)	CD3/CD28 stimulated resting CD4 T cells from aviremic donors are engrafted into humanized mice	Plasma HIV RNA detection (LTR primers) by qPCR	Charlins et al., 2017
MΦ-QVOA	Myeloid cells from SlV-infected macaques are stimulated with TNF-α; co-cultured with CEMX174 as targets	SIV gag RNA qPCR	Avalos et al., 2016