TEST:

Idylla™ BRAF Mutation Test

IdyllaTM testing proved to be a reliable and rapid alternative to NGS in the determination of BRAF V600 mutation. Although BRAF. status was available earlier, this had no influence on the treatment decision in most cases.

However, NGS had a 12-day longer TAT compared to EGFR PCR. Gene fusion-panel PCR and NGS identified an additional 11.6% of patients harboring actionable alterations who may benefit from FDA-approved targeted therapies.

Our results suggest that total loss of BRAF in patients with CRC is more likely to be observed in patients with KRAS mutation. A correlation with molecular BRAF status is necessary to definitively assess the benefit of BRAF IHC as an indicative method of BRAF and KRAS status in CRC which could be used as a first alternative in countries with low income.

Regardless, we recommend that Idylla BRAF cases with non-classical amplification curves undergo reflex NGS testing. These findings are likely relevant for other Idylla assays interrogating hotspot mutations in genes such as EGFR, IDH1/2, KRAS, and NRAS.

This report presents the initial analysis of the OPTIMEL study. The variant allele frequency of BRAF mutations in circulating tumor DNA is indicative of the objective response to BRAF and MEK inhibitors, as well as PFS. These findings could potentially influence our approach to managing patients with MM harboring a BRAF mutation.

BRAF V600E mutation was identified in 15 out of 40 secondary tumors with lymph node location (37.5%) and in 6 out of 15 secondary tumors with another location (40%) without statistically significant differences between the mutation frequency and the location of the secondary tumors. Our results support MM high genetic heterogeneity, pointing out the relationship between BRAF V600E mutation and several clinicopathological characteristics, in primary and metastatic MMs, stressing the importance of BRAF testing implementation in Romania.

"Biocartis...announces that it will host a free corporate workshop at the Annual Meeting of the ‘Association for Molecular Pathology’ (AMP), a leading molecular diagnostics conference, taking place between 14-18 November 2023 in Salt Lake City, Utah (US)."

Melanin bleaching can be a double-edged sword: The melanin may disappear, but the tissue's morphology and integrity of nuclear contents may be compromised. KMnOâ‚„ frequently compromises structural integrity and antigenicity in IHC. Thus, a gentler Hâ‚‚Oâ‚‚ treatment was selected for the molecular analysis of moderately/heavily pigmented melanomas.

The Idylla BRAF Mutation Assay is a highly reliable and sensitive platform for detecting BRAF pathogenic variants in codon 600 in malignant melanoma. The test can be performed in less than two hours, significantly improving turnaround time, thus allowing faster time to treatment.

The rapid demise of these two patients with BRAF V600E-mutant AML highlights their poor prognosis. Few cases of BRAF V600E-mutant AML have been described in the literature, all of which reported poor survival. Intriguingly, most cases exhibited monocytic morphology and concurrent KMT2A rearrangements, as was observed in case 2.

Patients RASmut on liquid biopsy at baseline are randomized to receive FOLFIRI plus Cetuximab or FOLFIRI plus Bevacizumab... These preliminary data suggest that liquid biopsy might better recapitulate the heterogeneity of mCRC and might be useful in clinical practice to complement tissue testing. The clinical relevance of RAS variants detected in cfDNA will be determined in the LIBImAb trial.

P=N/A | N=55 | Completed | Sponsor: Royal Cornwall Hospitals Trust | Recruiting ➔ Completed | N=200 ➔ 55

analysis for BRAFV600- and NRASQ61-mutant ctDNA on CSF using the Idylla® platform holds promise as a sensitive and specific complementary biomarker for the diagnosis of LMM, especially in cases with ambiguous imaging or CSF cytology results. The rapid (90 min) analysis of 1 ml of CSF, offering same day results, can be of important benefit, facilitating urgent treatment decisions.

P2 | "This is the first prospective randomized trial which evaluated anti-EGFR monoclonal antibody (panitumumab) in addition to standard of care (trifluridine-tipiracil) as third-line rechallenge therapy in chemo-refractory RASWT mCRC patients. Baseline liquid biopsy allows selection of RAS/BRAF WT ctDNA patients who could have a relevant clinical benefit. Clinical trial information: NCT05468892."

Beside established anti-CRC treatment, better understanding of the causality of CRC can be established by combining epidemiology and genetic/epigenetic on CRC etiology in Morocco. This approach may be able to significantly reduce the burden of disease in the country. Moreover, the Moroccan government should develop policy on CRC prevention and public health programs which may serve as a feasible setting to increase public awareness on lifestyle risk factors.

"The 23 common mutations detected in the BRAF and NRAS genes using this assay have significant impact on therapeutic selection. Somatic variants in the BRAF gene have been found in 37%-50% of all malignant melanomas, whereas somatic variants in the NRAS gene are found in 13%-25% of all melanomas. The Biocartis Idylla system offers a simple and rapid method to obtain somatic mutation information in these two genes that are of clinical importance."

Ras/Braf mutation frequency in our population approaches occidental series.

It is a minimally invasive method with a high degree of sensitivity and is representative of tumour heterogeneity. Future work is needed to evaluate cases with tissue-plasma discordance and the emerging monitoring of acquired mutations in RAS and BRAF as mechanisms of resistance.

Several clinicopathological factors were correlated with RAS-wt conversion, which may help select pts most likely to benefit from LB during treatment. Doing LB in pts with Ras-mut tumors should be considered, preferentially early in the disease-course, since loss of mutation represents an opportunity for treatment with anti-EGFR-therapy otherwise unlikely.

The primary endpoint is the PFS Plasma samples will be analyzed for KRAS, NRAS, and BRAF mutations by Idylla ctKRAS and Idylla ctNRAS-BRAF-EGFGR. All samples will be also analyzed by NGS, in order to better evaluate the correlation of tumor heterogeneity with pts’ outcomes.

"The Idylla BRAF test has the potential to make a significant positive impact on progression-free survival of malignant melanoma patients due to its rapid turnaround time. The Idylla BRAF test can be used as an adjunct to NGS for timely management of patients, particularly those with a poor performance status at presentation."

"The agreement rate was 91% (72/79). Although the agreement rate was high, we suggest that the use of IHC is more suitable for rapid BRAF testing on sentinel lymph node biopsies when associated with a low percentage and scattered tumor cells, which gave a high risk of non-contributory analysis and/or false negative results with the IdyllaBRAF Mutation Test."

"Our small sample size may be a limitation of the study. In our cohort, RAS-mutated tumours were associated with worse DFS and OS in early-stage CRC, whereas the remaining molecular variables had no prognostic influence."

In this article, we review the data about the fast and fully automated real-time PCR platform Idylla™ (Biocartis, Mechelen, Belgium) permitting the mutational analyses of BRAF, KRAS, NRAS, EGFR and microsatellite instability notably in melanoma, non-small-cell lung cancer and colorectal cancer samples. Future applications as well as the implementation of Idylla™ in the workflow of pathology and/or molecular biology laboratories are also discussed.

The aim of our study was to investigate the use of TPs on the Idylla system to reduce TATs, such that results would be available with routine surgical pathology reports. Our approach of collecting TP samples directly from tissue and testing them via the Idylla system significantly reduced potential TATs. Samples processed with rapid intent often provided results the same day as or before the pathology report release.

Our results demonstrate that the Idylla BRAF Mutation Assay produces accurate, precise results in less than 3 hours, with faster TAT compared to NGS. Single-use cartridges eliminate the need for manual sample preprocessing and the risk of PCR contamination. In addition, this sensitive assay correctly identifies BRAF V600 alterations in samples with necrosis and low tumor cell content.

The study has shown a 46% mutation rate for KRAS muta- tions in NSCLC samples, and with a KRAS G12C inhibitor on the hori- zon we have shown that it is feasible to use KRAS testing as a screening tool and for treatment stratification purposes in parallel. Results from the BRAF validation demonstrate the ability to detect mutations in the BRAF gene in a number of tissue types.

The liquid biopsy driven rechallenge strategy with anti-EGFR confirmed to be feasible in clinical practice. The clinical outcome resulted consistent with the literature data. In addition to the molecular selection through the analysis of ct-DNA for RAS, the long anti-EGFR free interval is confirmed as a prospective selection criterion for this therapeutic option.

Plasma-based, rapid ctDNA testing may be useful as an aid in detecting progression and gauging prognosis in patients with melanoma treated in the metastatic setting . The dynamics of ctDNA test positivity may indicate a need for more urgent imaging, particularly of the brain . Blood-based, semi-automated ctDNA detection may serve as an attractive adjunct to scheduled imaging surveillance in melanoma.

This study demonstrated that there may be changes in RAS/BRAF mutation from plasma analysis after progression in patients with mCRC . Since better survival in the patient group with wild type was detected compared to the RAS concordance group, the evaluation of RAS mutation status at the time of progression may be important in terms of disease prognosis and treatment options.

In details 11 patients (78.6%) underwent irinotecan+ cetuximab treatment, whereas 3 patients (21.4%) underwent panitumumab monotherapy . The rechallenge strategy with anti-EGFR confirmed to be feasible in clinical practice . The clinical outcome resulted consistent with the literature data . In addition to the molecular selection through the analysis of ct-DNA for RAS, previous response to anti EGFR treatment is confirmed as a prospective selection criteria for this therapeutic option.

"This included 2 cases of GG and 5 cases of PA. Our study found that BRAF V600E mutations are moderately frequent in PA and GG and that for these tumor entities, IHC VE1 is suitable for screening purposes, but all negative, equivocal, and weak positive cases should be further tested with molecular biology techniques, of which the Idylla system seems to be a promising tool."

P2, N=52, Active, not recruiting, Hoffmann-La Roche | Trial completion date: Jul 2021 --> Jul 2019