A novel flow cytometry-based cell capture platform for the detection, capture and molecular characterization of rare tumor cells in blood

Table 1 Mutation analysis of single or small groups of tumor cells

Cells	Template	EGFR mutation	Var. Freq. by Pyro.	Var. Freq. by MiSeq		Coverage min. = 10 × (212 amplicons)
Cells	Template	EGFR mutation	Var. Freq. by Pyro.	Var. Freq.	Total read	Coverage min. = 10 × (212 amplicons)
H1975	Unamplified gDNA	L858R (2573T>G)	75%	78.2%	13,070	100.0%
H1975	Unamplified gDNA	T790M (2369C>T)	74%	79.5%	11,044	100.0%
H1975	Amplified gDNA	L858R	73%	65.9%	372	96.7%
H1975	Amplified gDNA	T790M	72%	75.2%	10,013	96.7%
H1975	10 cells	L858R	71%	88.8%	4,479	92.0%
H1975	10 cells	T790M	79%	84.1%	27,539	92.0%
H1975	10 cells	L858R	73%	90.1%	601	93.5%
H1975	10 cells	T790M	77%	80.9%	35,070	93.5%
H1975	1 cell	L858R	68%	77.1%	109	90.6%
H1975	1 cell	T790M	79%	87.0%	31,312	90.6%
H1975	1 cell	L858R	69%	78.2%	368	80.2%
H1975	1 cell	T790M	61%	63.8%	27,341	80.2%
WBC from healthy donor	10 cells	L858R	2%	N/A	3,596	81.4%
WBC from healthy donor	10 cells	T790M	0%	N/A	18,497	81.4%
WBC from healthy donor	1 cell	L858R	3%	N/A	56	88.7%
WBC from healthy donor	1 cell	T790M	0%	N/A	256	88.7%

Footnote: Tumor cells or normal WBCs were fixed and stained followed by Ampli 1 WGA. WGA products were sequenced using a pyrosequencer or the Illumina MiSeq sequencer. Variant frequencies of two different mutations in the EGFR and the coverage distribution of WGA products using the TSACP are shown. Pyro.,psyrosequence Var. Freq., variant frequency. Coverage min., coverage minimum.

ISSN: 1479-5876