Plasma-SeqSensei™ (PSS) assays enable highly sensitive and quantitative detection of mutations in circulating tumour DNA (ctDNA) from the blood plasma of cancer patients by utilising next-generation sequencing technology. PSS's short and standardised workflow delivers results within two days including the generation of easy-to-read reports using laptop-compatible Plasma-SeqSensei™ IVD Software.

The kit covers key gene mutations such as BRAF, EGFR, KRAS, NRAS and PIK3CA to detect established and emerging predictive markers, resistance mutations, and frequently occurring genetic alterations in cancer.

PSS Solid Cancer IVD Kit is intended to support clinicians with:

detecting minimal residual disease
recurrence surveillance
(neo-)adjuvant response monitoring
analysing the RAS mutation status to determine the potential benefit of anti-epidermal growth factor receptor (EGFR) therapy for colorectal cancer patients

Features

PSS technology

Confident mutant reporting

To increase sensitivity and reduce error rates associated with sequencing, the PSS library preparation workflow employs SafeSEQ technology ¹. This technology assigns a unique identifier (UID) to each DNA molecule during the target selection step. UIDs help track and identify real mutations in samples and distinguish them from polymerase or sequencing errors. PSS detects 0.07% and higher mutant allele fractions (MAF) with 95% certainty in a background of 10,000 wild-type copies.

Beyond MAF – absolute mutant quantification

PSS technology utilises internal quantifier Quantispike, which allows robust quantification of tumour-specific sequences over a broad dynamic range down to a limit of detection (LOD) of seven mutant molecules. Additionally, absolute quantification reported as a number of mutant molecules detected in the tested volume of blood helps to facilitate a meaningful comparison of changes in ctDNA levels over time. 

Plasma-SeqSensei™ IVD Software

PSS IVD Software offers the possibility to effortlessly plan and analyse sequencing runs and report data easily. It is the first IVD-certified software designed to run on an office PC or laptop – no workstation or server structure is needed. The interface is divided into three functional modules: run planning, data analysis and reporting. The automatically generated report includes sample metrics and mutation status, both listing MAFs as well as the absolute number of mutant molecules detected in the sample volume, alongside sequencing quality metrics.

References

Kinde, I., Wu, J., Papadopoulos, N., Kinzler, K. W., & Vogelstein, B. (2011). Detection and quantification of rare mutations with massively parallel sequencing. Proceedings of the National Academy of Sciences of the United States of America, 108(23), 9530–9535. https://doi.org/10.1073/pnas.1105422108

Technical Specifications

Feature	Plasma-SeqSensei^TM Solid Cancer IVD Kit
Sample type	Whole blood and plasma
Sample capacity	2 – 16 samples per kit
QC function	Positive control and no template control (NTC) applied to every run
Compatible platforms	Illumina NextSeq 500/550™
Input DNA required	5.7–95 ng/116 µl
Number of amplicons	17
Sensitivity	0.07% allele frequency at 95% certainty at 10,000 wildtype copies
Cut-off	7 mutant molecules
Cat. No.	ZR150534

Gene targets

Gene	Transcript*	Coding sequence start	Coding sequence end	Most frequent mutations detected (AA change)
BRAF	ENST00000288602	1,383	1,431	G469A/R/V/E, G466V/E
BRAF	ENST00000288602	1,742	1,813	V600E/K/R/M, K601E, D594G
EGFR	ENST00000275493	2,116	2,177	G719A/S
EGFR	ENST00000275493	2,565	2,620	L858R, L861Q
EGFR	ENST00000275493	2,225	2,279	E746_A750del, L747_P753delinsS, L747_T751del, L747_A750delinsP, E746_S752delinsV
EGFR	ENST00000275493	2,361	2,403	T790M
EGFR	ENST00000275493	2,284	2,325	S7681I
KRAS	ENST00000256078	419	445	A146T/V
KRAS	ENST00000256078	326	352	K117N
KRAS	ENST00000256078	34	102	G12D/V/C/A/S/R/F, G13D/C/R/V/A
KRAS	ENST00000256078	169	102	Q61H/R/L/H/K, A59T
NRAS	ENST00000369535	162	210	Q61R/K/L/H
NRAS	ENST00000369535	420	449	A146V/T
NRAS	ENST00000369535	1	52	G12D/C/S/A/V/R, G13R/V/C/S
NRAS	ENST00000369535	341	364	L120D, K117R
PIK3CA	ENST00000263967	3,118	3,195	H1047R/L/Y, G1049R, M1043I/V
PIK3CA	ENST00000263967	1,611	1,659	E545K/A/G/Q, E542K, Q546K/R/P