Table 3.

Prospective molecular diagnosis of LM cyst fluid cfDNA with multiplex ddPCR.

						Multiplex ddPCR		Singleplex ddPCR
Patient	Sex	Agea	Location	Cyst fluid volume (mL)	PIK3CA Variant	VAF (%)b	Droplets (Var/WT)	VAF (%)b	Droplets (Var/ WT)
LR16-265c	F	11	Axilla	6.0	p.H1047R (c.3140A>G)	1.4	124/8172	1.4	170/10972
LR19-442	M	6	Retro-peritoneum	9.0	p.E545K (c.1633G>A)	0.2	27/12098	0.2	20/11395
LR19-443	M	1 mo	Axilla	2.0	NEG	NEG	- /30632	-	-
LR19-446	M	9 mo	Chest wall	5.0	p.E542K (c.1624G>A)	6.5	920/12116	6.7	856/10878
LR19-481	M	9 mo	Neck, lower face	6.5d	p.E545K (c.1633G>A)	1.6	152/8910	1.5	162/9662

Abbreviations: cfDNA – cell free DNA, mo – months old, mL – milliliters, NEG – no variant detected, VAF – variant allele fraction, Var – variant, WT – wild-type

^aAge at first cyst fluid collection, in years unless otherwise specified.

^bVAF calculated using droplet concentrations and only reported for samples in which sample variant concentration was statistically different from WT cfDNA control variant concentration based on 95% total error confidence intervals.

^cPatient previously reported.¹¹ No variant was found in previous publication when DNA was extracted from 18g needle biopsy of lesion tissue taken at the time of sclerotherapy treatment.

^dSample collected in Streck tube.