Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen® FFPE BRAF qPCR assay
Mutations in BRAF exon 15 lead to conformational changes in its activation loops, resulting in constitutively active BRAF proteins which are implicated in the development of several human cancer types. Different BRAF inhibitors have been developed and introduced in clinical practice. Identification...
| Main Authors: | , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2023-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9910728/?tool=EBI |
| _version_ | 1811166144723156992 |
|---|---|
| author | Anna Lahn Sørensen Mariann Guldmann-Christensen Michael Børgesen Rasmus Koefoed Petersen Katharina Flugt Julie Mejer Holmgaard Duelund Majbritt Hauge Kyneb Jan Lorenzen Emma Pipó-Ollé Samantha Epistolio Alice Riva Giulia Dazio Elisabetta Merlo Tine Meyer Ulf Bech Christensen Milo Frattini |
| author_facet | Anna Lahn Sørensen Mariann Guldmann-Christensen Michael Børgesen Rasmus Koefoed Petersen Katharina Flugt Julie Mejer Holmgaard Duelund Majbritt Hauge Kyneb Jan Lorenzen Emma Pipó-Ollé Samantha Epistolio Alice Riva Giulia Dazio Elisabetta Merlo Tine Meyer Ulf Bech Christensen Milo Frattini |
| author_sort | Anna Lahn Sørensen |
| collection | DOAJ |
| description | Mutations in BRAF exon 15 lead to conformational changes in its activation loops, resulting in constitutively active BRAF proteins which are implicated in the development of several human cancer types. Different BRAF inhibitors have been developed and introduced in clinical practice. Identification of BRAF mutations influences the clinical evaluation, treatment, progression and for that reason a sensitive and specific identification of BRAF mutations is on request from the clinic. Here we present the SensiScreen® FFPE BRAF qPCR Assay that uses a novel real-time PCR-based method for BRAF mutation detection based on PentaBases proprietary DNA analogue technology designed to work on standard real-time PCR instruments. The SensiScreen® FFPE BRAF qPCR Assay displays high sensitivity, specificity, fast and easy-to-use. The SensiScreen® FFPE BRAF qPCR Assay was validated on two different FFPE tumour biopsy cohorts, one cohort included malignant melanoma patients previously analyzed by the Cobas® 4800 BRAF V600 Mutation Test, and one cohort from colorectal cancer patients previously analyzed by mutant-enriched PCR and direct sequencing. All BRAF mutant malignant melanoma patients were confirmed with the SensiScreen® FFPE BRAF qPCR Assay and additional four new mutations in the malignant melanoma cohort were identified. All the previously identified BRAF mutations in the colorectal cancer patients were confirmed, and additional three new mutations not identified with direct sequencing were detected. Also, one new BRAF mutation not previously identified with ME-PCR was found. Furthermore, the SensiScreen® FFPE BRAF qPCR Assay identified the specific change in the amino acid. The SensiScreen® FFPE BRAF qPCR Assay will contribute to a more specific, time and cost saving approach to better identify and characterize mutations in patients affected by cancer, and consequently permits a better BRAF characterization that is fundamental for therapy decision. |
| first_indexed | 2024-04-10T15:47:47Z |
| format | Article |
| id | doaj.art-bfdf2cd7b7064edb96b70276f9e039e1 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-10T15:47:47Z |
| publishDate | 2023-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-bfdf2cd7b7064edb96b70276f9e039e12023-02-12T05:31:11ZengPublic Library of Science (PLoS)PLoS ONE1932-62032023-01-01182Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen® FFPE BRAF qPCR assayAnna Lahn SørensenMariann Guldmann-ChristensenMichael BørgesenRasmus Koefoed PetersenKatharina FlugtJulie Mejer Holmgaard DuelundMajbritt Hauge KynebJan LorenzenEmma Pipó-OlléSamantha EpistolioAlice RivaGiulia DazioElisabetta MerloTine MeyerUlf Bech ChristensenMilo FrattiniMutations in BRAF exon 15 lead to conformational changes in its activation loops, resulting in constitutively active BRAF proteins which are implicated in the development of several human cancer types. Different BRAF inhibitors have been developed and introduced in clinical practice. Identification of BRAF mutations influences the clinical evaluation, treatment, progression and for that reason a sensitive and specific identification of BRAF mutations is on request from the clinic. Here we present the SensiScreen® FFPE BRAF qPCR Assay that uses a novel real-time PCR-based method for BRAF mutation detection based on PentaBases proprietary DNA analogue technology designed to work on standard real-time PCR instruments. The SensiScreen® FFPE BRAF qPCR Assay displays high sensitivity, specificity, fast and easy-to-use. The SensiScreen® FFPE BRAF qPCR Assay was validated on two different FFPE tumour biopsy cohorts, one cohort included malignant melanoma patients previously analyzed by the Cobas® 4800 BRAF V600 Mutation Test, and one cohort from colorectal cancer patients previously analyzed by mutant-enriched PCR and direct sequencing. All BRAF mutant malignant melanoma patients were confirmed with the SensiScreen® FFPE BRAF qPCR Assay and additional four new mutations in the malignant melanoma cohort were identified. All the previously identified BRAF mutations in the colorectal cancer patients were confirmed, and additional three new mutations not identified with direct sequencing were detected. Also, one new BRAF mutation not previously identified with ME-PCR was found. Furthermore, the SensiScreen® FFPE BRAF qPCR Assay identified the specific change in the amino acid. The SensiScreen® FFPE BRAF qPCR Assay will contribute to a more specific, time and cost saving approach to better identify and characterize mutations in patients affected by cancer, and consequently permits a better BRAF characterization that is fundamental for therapy decision.https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9910728/?tool=EBI |
| spellingShingle | Anna Lahn Sørensen Mariann Guldmann-Christensen Michael Børgesen Rasmus Koefoed Petersen Katharina Flugt Julie Mejer Holmgaard Duelund Majbritt Hauge Kyneb Jan Lorenzen Emma Pipó-Ollé Samantha Epistolio Alice Riva Giulia Dazio Elisabetta Merlo Tine Meyer Ulf Bech Christensen Milo Frattini Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen® FFPE BRAF qPCR assay PLoS ONE |
| title | Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen® FFPE BRAF qPCR assay |
| title_full | Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen® FFPE BRAF qPCR assay |
| title_fullStr | Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen® FFPE BRAF qPCR assay |
| title_full_unstemmed | Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen® FFPE BRAF qPCR assay |
| title_short | Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen® FFPE BRAF qPCR assay |
| title_sort | detection of braf mutations in malignant melanoma and colorectal cancer by sensiscreen r ffpe braf qpcr assay |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9910728/?tool=EBI |
| work_keys_str_mv | AT annalahnsørensen detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT mariannguldmannchristensen detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT michaelbørgesen detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT rasmuskoefoedpetersen detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT katharinaflugt detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT juliemejerholmgaardduelund detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT majbritthaugekyneb detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT janlorenzen detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT emmapipoolle detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT samanthaepistolio detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT aliceriva detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT giuliadazio detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT elisabettamerlo detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT tinemeyer detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT ulfbechchristensen detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay AT milofrattini detectionofbrafmutationsinmalignantmelanomaandcolorectalcancerbysensiscreenffpebrafqpcrassay |