Table 4.

Estimated Extra-Poisson Variation for Effect of Cryopreservation for Classic Quantitative Viral Outgrowth Assay Versus Next-Generation Assays, After Adjustment for Fixed Scale Differences Between Assays at the Aliquot, Batch, and Assay Levels

Level of Analysis	Estimated Extra-Poisson Variation, Posterior Median Fold Change (95% CI), for Classic QVOA Versus Next-Generation Assays
	QVOA RNA	QVOA Simoa	TILDA	caRNA1	caRNA2	cfRNA
Aliquot level	1.5 (1.1–2.2)	1.4 (1.0–2.0)	1.6 (1.3–2.2)	2.6 (2.1–3.4)	1.9 (1.6–2.5)	1.7 (1.4–2.1)
Batch level (alone)	1.7 (1.0–2.4)	2.1 (1.3–3.4)	1.2 (1.0–2.0)	1.0 (1.0–1.8)	1.2 (1.0–2.1)	1.2 (1.0–1.9)
Aliqot + batch	2.0 (1.6–2.7)	2.3 (1.6–3.6)	1.7 (1.4–2.5)	2.6 (2.2–3.5)	2.1 (1.7–2.8)	1.8 (1.5–2.3)
Assay level (alone)	1.5 (1.0–2.5)	1.4 (1.0–2.8)	2.0 (1.0–4.0)	1.5 (1.0–2.7)	2.0 (1.0–3.9)	1.4 (1.0–2.3)
Aliquot + batch + assay	2.3 (1.8–3.4)	2.5 (1.8–4.3)	2.5 (1.7–4.6)	2.9 (2.3–4.4)	2.8 (1.9–4.9)	2.0 (1.6–2.9)
Assay type	1.0 (1.0–2.4)	1.0 (1.0–2.9)	1.0 (1.0–7.1)	1.0 (1.0–3.0)	1.0 (1.0–5.8)	1.0 (1.0–2.3)
Aliquot + batch + assay + assay type	2.4 (1.8–4.0)	2.7 (1.8–5.0)	3.1 (2.1–8.0)	3.0 (2.3–5.5)	3.2 (2.3–7.6)	2.0 (1.6–3.5)
Frozen effect, %	−31 (−62 to 30)	−41 (−80 to 71)	−27 (−60 to 26)	−8 (−58 to 96)	−12 (−52 to 63)	−52 (−72 to −22)

Abbreviations: caRNA1, cell-associated human immunodeficiency virus (HIV) gag RNA; caRNA2, cell-associated HIV tat-rev RNA; cfRNA, cell-free RNA; CI, credible interval; NA, not applicable; QVOA, quantitative viral outgrowth assay; QVOA RNA, QVOA by University of California, San Diego, with HIV RNA readout; QVOA Simoa, QVOA by Southern Research using Simoa readout; TILDA, tat/rev-induced limiting dilution assay.