TABLE 2.

Pathogenic RB1 Variants and Cell-Free DNA Concentrations for All 26 Diagnostic Aqueous Humor Samples

Case ID_Eye if Bilateral	gDNA RB1 Variant	AH RB1 Variants	% VAF	Total Reads, Altered Reads	95% CI	dsDNA (ng/μL)
33	Negative	c.2325+ 1G>A	99.7	957, 954	99.0–100.0	3.4
44_OD	c.1666C>T	c.1666C>T	67.2	64, 43	53.6–80.8	1.2
44_OS	c.1666C>T	c.1666C>T	98.7	947, 935	97.4–99.7	2.3
—	—	RB1 gene deletion	—	—	—	—
45	Negative	c.1422–1G>A	97.9	330,323	95.5–100.0	0.6
46	Negative	c.1363C>T	99.4	359,357	93.3–98.1	0.5
47	Negative	c.958C>T	93.5	31, 29	73.1–99.7	3.0
48	Negative	ND	ND	ND	—	4.1
51	Negative	Loss of 13q	—	—	—	n/a
55	Negative	c.869dup	43.3	30, 13	32.3–72.4	n/a
—	—	c.1215+1G>A	40.8	71, 29	33.0–58.0	n/a
64	Heterozygous deletion of exons 24–27	RB1 gene deletion	—	—	—	n/a
65	Negative	c.1589_1590del	66.7	6, 4	20.6–98.3	24.0
—	—	c.2330dup	33.3	6, 2	3.9–76.2	24.0
66	c.1362 C>G	c.380G>T	50.0	42, 21	38.6–69.2	46.4
—	—	c.1362C>G	30.5	59, 18	20.2–47.4	46.4
67	Negative	c.1028_1029del	40.4	104,42	36.0–53.0	0.5
—	—	c.1794_1801dup	39.6	159,63	39.3–49.7	0.5
70_OD	c.607+ 1G>C	ND	ND	ND	—	ND
70_OS	c.607+ 1G>C	ND	ND	ND	—	0.5
72_OD	c.1399C>T	ND	ND	ND	—	ND
72_OS	c.1399C>T	c.571G >A	100	24, 24	—	56.6
73	Negative	c.1333–5_1333del	97.8	89, 87	87.6–97.2	27.8
74_OD	c.1215+1G >A	ND	ND	ND	—	1.1
74_OS	c.1215+1G >A	c.1215+1G>A	100	42, 42	—	1.5
76	Negative	c.182_183del	44.6	56, 25	30.0–53.9	0.1
—	—	c.1072C>T	42.9	35, 15	26.1–64.8	0.1
—	—	c.751C >T	97.5	237, 231	93.0–97.4	0.1
78	c.958C>T	c.958C >T	97.6	42, 41	90.1–98.8	0.4
83	Negative	ND	ND	ND	—	0.5
84	Negative	ND	ND	ND	—	0.1
88_OD	c.1996dup	c.1996dup	100	63, 63	—	0.6
88_OS	c.1996dup	c.1996dup	93.2	74, 69	87.3–96.3	0.3

AH = aqueous humor, dsDNA = double-stranded DNA; gDNA = genomic DNA, n/a = not available, ND = not detected, VAF = variant allele frequency.