Detection of microsatellite instability with Idylla MSI assay in colorectal and endometrial cancer

Ukkola, Iiris; Nummela, Pirjo; Pasanen, Annukka; Lepistö, Anna; Kytölä, Soili; Bützow, Ralf; Ristimäki, Ari

doi:10.1007/s00428-021-03082-w

Cited by 28 publications

(33 citation statements)

References 26 publications

(28 reference statements)

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“…KRAS and NRAS mutations were identified, and the BRAFV600wt status confirmed, from the 62 CRC FFPE samples using real-time PCR-based CE-IVD-validated Idylla KRAS and NRAS-BRAF mutation assays (Biocartis NV, Mechelen, Belgium). Furthermore, the MSI status of the dMMR samples was confirmed with Idylla MSI test that was performed as described previously [10]. The Idylla KRAS Mutation Test detects 21 KRAS mutations in exons 2, 3 and 4, whereas the Idylla NRAS-BRAF Mutation Test detects 18 NRAS mutations in exons 2, 3 and 4, and five BRAF mutations in codon 600.…”

Section: Idylla Kras and Nras-braf Mutation Tests And Msi Testmentioning

confidence: 99%

“…For immunohistochemical analyses, 4-µm sections cut from the FFPE CRC tissue blocks (n = 62) were used. The MMR IHC was performed as described previously [10] and BRAFV600E mutation was detected by using the specific monoclonal ready-to-use antibody (clone VE1, 760-5095, Roche, Tucson, AZ), utilising detection with OptiView DAB kit (760-700, Roche) and additional Amplification Kit (760-099, Roche) [12]. Pan-TRK IHC was performed using ready-to-use monoclonal antibody (clone EPR17341, 790-7026, Roche) and detection by OptiView DAB kit (760-700, Roche).…”

Section: Immunohistochemistrymentioning

confidence: 99%

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Gene fusions and oncogenic mutations in MLH1 deficient and BRAFV600E wild-type colorectal cancers

et al. 2022

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Gene fusions can act as oncogenic drivers and offer targets for cancer therapy. Since fusions are rare in colorectal cancer (CRC), their universal screening seems impractical. Our aim was to investigate gene fusions in 62 CRC cases with deficient MLH1 (dMLH1) and BRAFV600E wild-type (wt) status from a consecutive real-life series of 2079 CRCs. First, gene fusions were analysed using a novel FusionPlex Lung v2 RNA–based next-generation sequencing (NGS) panel, and these results were compared to a novel Idylla GeneFusion assay and pan-TRK immunohistochemistry (IHC). NGS detected seven (7/62, 11%) NTRK1 fusions (TPM3::NTRK1, PLEKHA6::NTRK1 and LMNA::NTRK1, each in two cases, and IRF2BP2::NTRK1 in one case). In addition, two ALK, four RET and seven BRAF fusions were identified. Idylla detected seven NTRK1 expression imbalances, in line with the NGS results (overall agreement 100%). Furthermore, Idylla detected the two NGS–identified ALK rearrangements as one specific ALK fusion and one ALK expression imbalance, whilst only two of the four RET fusions were discovered. However, Idylla detected several expression imbalances of ALK (n = 7) and RET (n = 1) that were found to be fusion negative with the NGS. Pan-TRK IHC showed clearly detectable, fusion partner-dependent staining patterns in the seven NTRK1 fusion cases. Overall agreement for pan-TRK antibody clone EPR17341 was 98% and for A7H6R 100% when compared to the NGS. Of the 62 CRCs, 43 were MLH1 promoter hypermethylated (MLH1ph) and 39 were RASwt. All fusion cases were both MLH1ph and RASwt. Our results show that kinase fusions (20/30, 67%) and most importantly targetable NTRK1 fusions (7/30, 23%) are frequent in CRCs with dMLH1/BRAFV600Ewt/MLH1ph/RASwt. NGS was the most comprehensive method in finding the fusions, of which a subset can be screened by Idylla or IHC, provided that the result is confirmed by NGS.

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Section: Idylla Kras and Nras-braf Mutation Tests And Msi Testmentioning

confidence: 99%

Section: Immunohistochemistrymentioning

confidence: 99%

Gene fusions and oncogenic mutations in MLH1 deficient and BRAFV600E wild-type colorectal cancers

et al. 2022

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“…of 16 patients with different types of solid tumors, including breast cancer [49,50], who may benefit from ICIs.…”

Section: Institutional Review Board Statementmentioning

confidence: 99%

“…Generally, the most widely adopted procedure for MSI evaluation relies on the Sanger sequencing-based Bethesda panel, which covers two mononucleotide (BAT-25 and BAT-26) and three dinucleotide (D5S346, D2S123, and D17S250) repetitions [13] or, alternatively, a panel covering five poly-A mononucleotide repeats (BAT-25, BAT-26, NR-21, NR-24, NR-27) [14]. However, recent studies have demonstrated that the fully automated PCR high-resolution melt curve analysis (Idylla TM , Biocartis, Mechelen, Belgium) [11,[15][16][17][18][19][20][21][22][23][24][25][26] and the automated microfluidic electrophoretic run chip-based assay (TapeStation 4200, Agilent Technologies, Santa Clara, CA, USA) are easier to use, faster, and less expensive than Sanger sequencing [11,27,28].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Micro Satellite Instability and Mismatch Repair Status in Different Solid Tumors: A Multicenter Analysis in a Real World Setting

Malapelle

Parente

Pepe

et al. 2021

Cells

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Immune-checkpoint inhibitors (ICIs) play a key role in the treatment of advanced stage colorectal cancer (CRC) patients featuring a deficient DNA mismatch repair (dMMR) system or a high microsatellite instability (MSI-H) profile. However, beyond the established role in CRC patients, ICIs have highly proven efficacy in other solid tumors featuring MSI-H/dMMR status represented by endometrial, gastric, ovarian, prostatic, and pancreatic carcinomas (EC, GC, OC, PrC, and PaC). Our aim was to compare the concordance rates among the Idylla™ MSI test, TapeStation 4200, and immunohistochemical (IHC) analysis in assessing MSI-H/dMMR status in EC, GC, OC, PrC, and PaC patients. The Sanger sequencing-based Titano MSI test was used in discordant cases. One hundred and eighty-five cases (n = 40 PrC, n = 39 GC, n = 38 OC, n = 35 PaC, and n = 33 EC) were retrospectively selected. MMR protein expression was evaluated by IHC. After DNA quality and quantity evaluations, the IdyllaTM and TapeStation 4200 platforms were adopted for the evaluation of MSI status. Remarkably, compared to IHC, the Idylla™ platform achieved a global concordance rate of 94.5% (154/163) for the microsatellite stable (MSS)/proficient MMR (pMMR) cases and 77.3% (17/22) for the MSI-H/dMMR cases. Similarly, a global concordance rate of 91.4% (149/163) and 68.2% (15/22) for MSS/pMMR and MSI-H/dMMR cases was also identified between IHC and the TapeStation 4200 microfluidic system. In addition, a global concordance of 93.1% (148/159) and 69.2% (18/26) for MSS/pMMR and MSI-H/dMMR cases was observed between the Idylla™ and TapeStation 4200 platforms. Discordant cases were analyzed using the Titano MSI kit. Overall, our data pinpointed a central role for molecular techniques in the diagnostic evaluation of dMMR/MSI-H status not only in CRC patients but also in other types of solid tumors.

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“…Molecular MSI analysis is also used, but as a disadvantage, this technique does not provide information on the specific MMR gene to be tested. Further, its sensitivity may be lower given the relatively high frequency of MSH6 loss in EC and the typically lower level of MSI caused by the inactivation of this particular gene [ 8 , 9 ]. As only 10% of all MMR deficient ECs are associated with germline mutation, immunohistochemistry alone is not enough to diagnose LS [ 1 ].…”

Section: Introductionmentioning

confidence: 99%

Testing for Lynch Syndrome in Endometrial Carcinoma: From Universal to Age-Selective MLH1 Methylation Analysis

Pasanen

Loukovaara

Kaikkonen

et al. 2022

Cancers

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International guidelines recommend universal screening of endometrial carcinoma (EC) patients for Lynch syndrome (LS). This screening is generally based on mismatch repair (MMR) protein immunohistochemistry followed by MLH1 methylation analysis of MLH1-negative cases to exclude the likely sporadic cases from germline testing. As LS-associated EC is uncommon in the elderly, age-selective methylation testing could improve cost-efficiency. We performed MMR immunohistochemistry on 821 unselected ECs (clinic-based cohort) followed by a MLH1 promoter methylation test of all MLH1/PMS2-negative tumors. Non-methylated MLH1-deficient cases underwent NGS and MLPA-based germline analyses to identify MLH1 mutation carriers. A reduction in the test burden and corresponding false negative rates for LS screening were investigated for various age cut-offs. In addition, the age distribution of 132 MLH1 mutation carriers diagnosed with EC (registry-based cohort) was examined. A germline MLH1 mutation was found in 2/14 patients with non-methylated MLH1-deficient EC. When compared to a universal methylation analysis, selective testing with a cut-off age of 65 years, would have reduced the testing effort by 70.7% with a false negative rate for LS detection of 0% and 3% in the clinic and registry-based cohorts, respectively. The use of age-selective methylation analysis is a feasible way of reducing the costs and laboratory burden in LS screening for EC patients.

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Detection of microsatellite instability with Idylla MSI assay in colorectal and endometrial cancer

Cited by 28 publications

References 26 publications

Gene fusions and oncogenic mutations in MLH1 deficient and BRAFV600E wild-type colorectal cancers

Gene fusions and oncogenic mutations in MLH1 deficient and BRAFV600E wild-type colorectal cancers

Evaluation of Micro Satellite Instability and Mismatch Repair Status in Different Solid Tumors: A Multicenter Analysis in a Real World Setting

Testing for Lynch Syndrome in Endometrial Carcinoma: From Universal to Age-Selective MLH1 Methylation Analysis

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