APIS ESR1 Mutations Kit

The APIS ESR1 Mutations Kit is an advanced qPCR assay for the sensitive and precise detection of mutations within the oestrogen receptor gene. The ESR1 Mutations Kit is a qualitative test, detecting eleven ESR1 mutations across three exons: exon 5 (E380Q), exon 7 (S463P) and exon 8 (P535H, L536R, L536Q, L536H, L536P, Y537C, Y537S, Y537N and D538G).

The kit includes mutation-specific probes to enable highly sensitive detection of the target mutations. Additionally, it utilises PCR clamp and blocker technology, which ensures specific amplification of the mutant sequence, even in the presence of a high wild-type background.

It is suitable for use in both centralised and decentralised settings, especially for those facilities that require a testing solution without the need for additional specialised equipment e.g. NGS.

Benefits of Using the APIS ESR1 Mutations Kit

Wide target coverage – the kit has been designed to cover 11 different ESR1 mutations>
High specificity – includes clamp and blocking technology to ensure no wild-type detection
High sensitivity – all mutations are detected at ≤ 1% MAF (Mutant Allele Frequency)
Easy to use – our assays are designed to be user-friendly, with a simple protocol. The reagents have been optimised for precise and sensitive detection in human DNA.

Assay Mutations and analytical sensitivity in contrived samples

The APIS ESR1 Mutations Kit detects 11 ESR1 mutations. Contrived samples of 0.008–1.6% mutant allele frequency (MAF) were prepared with a total copy of 5000 per reaction. The LoD is based on the highest mutant allele frequency with 95% correct calls observed for each target in 24 replicates. The claimed LoD for the targets in % MAF is ≤ 1% for all targets (Table 1).

Table 1. 11 ESR1 mutations detected in the APIS ESR1 Mutations Kit. LoD in % MAF and copies (cp). The mutant copies at LoD and WT copies at LoD describe the copies of each DNA fragment in the contrived LoD samples.

Supporting Data for the APIS ESR1 Mutations Kit

Linearity

An 8-level dilution series of DNA fragments specific to each mutation were used to evaluate PCR efficiency and R-sq values (Table 2 and Figure 1).

Table 2. Linearity analysis of each target in the APIS ESR1 Mutations Kit.

Figure 1. Linear regression of the D538G mutation across the 8-level dilution series.

APIS ESR1 Mutations Kit Sensitivity & Selectivity

APIS ESR1 Mutations Kit performance with SensID ESR1 Reference Set 1% AF cfDNA (SID-000144)

Highly sensitive and specific detection of SensID ESR1 Mutation Reference Standards at 1% MAF

APIS ESR1 Mutations Kit Specifications

The kit has undergone testing as part of APIS’ unique assay development pipeline, which includes in silico and wet laboratory testing. As a result, researchers can have confidence in the accuracy, sensitivity, specificity, and reliability of the kit. The qPCR assay’s efficiency and linearity were assessed using contrived synthetic DNA samples.

APIS ESR1 and PIK3CA Mutations Kits Bundle - Achieve ≤ 0.5% MAF Detection

Discover the advanced capabilities of the APIS ESR1 and PIK3CA Mutation Kits, both designed to achieve a ≤ 0.5% Limit of Detection (LoD).

The APIS ESR1 Mutations Kit is an advanced qPCR assay for the sensitive and precise detection of mutations within the oestrogen receptor gene (ESR1). The APIS ESR1 Mutations Kit is a qualitative test, detecting eleven ESR1 mutations from DNA extracted from plasma (cfDNA).

The APIS PIK3CA Mutations Kit is a real-time qualitative PCR test for the detection of the five most prevalent activating mutations in the PIK3CA gene, from DNA extracted from formalin-fixed, paraffin-embedded (FFPE) breast tumour tissue (gDNA) or blood plasma (cfDNA).

For the APIS ESR1 Mutations Kit, additional testing with samples at 0.5% MAF has demonstrated that the kit can achieve mutation detection at ≤ 0.5% MAF with a wild-type (WT) background of 10,000 or 50,000 copies. This detection capability applies to the following mutations: E380Q, D538G, Y537S, Y537C, L536H, L536Q, Y537N, and P535H, each reaching ≤ 0.5% MAF sensitivity.

Mutations Detected by the APIS ESR1 Mutations Kit and APIS PIK3CA Mutations Kit

Table 3. The different mutations detected by both the ESR1 Mutations Kit and the PIK3CA Mutations Kit

APIS ESR1 Mutations Kit - Available now

The APIS ESR1 Mutations Kit is currently available as a research use only (RUO) product. Not applicable for use in diagnostic procedures.

For more information on this product please contact us and select ‘ESR1 Mutations Kit’

Ordering Information

Product	Test Type	Kit Size	Catalogue Number	Price

APIS ESR1 Mutations Kit	RUO	24 samples plus controls	02201 (distributed by APIS)	Available upon request
APIS ESR1 Mutations Kit	RUO	24 samples plus controls	02202 (distributed by Biocartis)	Available upon request
APIS ESR1 Mutations Controls	RUO	24 samples plus controls	02801 (distributed by APIS)	Available upon request
APIS ESR1 Mutations Controls	RUO	24 samples plus controls	02802 (distributed by Biocartis)	Available upon request