{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T13:02:36Z","timestamp":1785416556075,"version":"3.56.0"},"reference-count":26,"publisher":"IMR Press","issue":"7","license":[{"start":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T00:00:00Z","timestamp":1784764800000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Agena Biosciences"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Front. Biosci. (Landmark Ed)"],"abstract":"<jats:p>Background:Accurate identification of actionable somatic variants is essential for therapeutic stratification in colorectal cancer (CRC). While next-generation sequencing (NGS) enables comprehensive genomic profiling, targeted approaches may provide faster and more practical alternatives for routine diagnostics.Methods:This study evaluated the analytical performance of the Agena Bioscience iPLEX\u00ae High Sensitivity (HS) Colon Panel using MassARRAY MALDI-TOF technology for detection of hotspot variants in KRAS, NRAS, BRAF, and PIK3CA from formalin-fixed paraffin-embedded (FFPE) specimens. A total of 60 unique clinical and reference samples were analyzed, targeting 26 single-nucleotide variants and compared with an orthogonal targeted NGS assay. Limit of detection (LOD) was assessed using serial dilutions of reference materials, and intra- and inter-run reproducibility was evaluated across multiple runs. Analytical performance metrics including positive and negative percent agreement, predictive values, and error rates were calculated.Results:The assay demonstrated complete concordance with NGS across all evaluated variants, requiring only 20 ng of DNA input, compared to 80\u2013120 ng for a successful NGS run. LOD studies showed reliable detection of multiple variants at approximately 5% variant allele frequency, with BRAF p.V600E detectable near 1%. Intra- and inter-run analyses achieved 100% concordance, confirming assay reproducibility. Aggregated performance metrics demonstrated high sensitivity and specificity across a heterogeneous sample set.Conclusions:These findings establish the iPLEX\u00ae HS Colon Panel as a reliable platform for rapid detection of clinically actionable hotspot mutations. This study represents analytical validation using mixed FFPE tumor specimens; evaluation in larger colorectal cancer cohort, is warranted.<\/jats:p>","DOI":"10.31083\/fbl53019","type":"journal-article","created":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T12:17:49Z","timestamp":1785413869000},"source":"Crossref","is-referenced-by-count":0,"title":["Performance Evaluation of a Colorectal Cancer-Associated MassARRAY-Based Hotspot Genotyping Assay for KRAS, NRAS, BRAF, and PIK3CA Using Solid Tumor Specimens"],"prefix":"10.31083","volume":"31","author":[{"given":"Vishakha","family":"Vashisht","sequence":"first","affiliation":[{"name":"Department of Pathology, Augusta University, Augusta, GA 30909, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashutosh","family":"Vashisht","sequence":"additional","affiliation":[{"name":"Department of Pathology, Augusta University, Augusta, GA 30909, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashis K.","family":"Mondal","sequence":"additional","affiliation":[{"name":"Department of Pathology, Augusta University, Augusta, GA 30909, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pankaj K.","family":"Ahluwalia","sequence":"additional","affiliation":[{"name":"Department of Pathology, Augusta University, Augusta, GA 30909, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jaspreet","family":"Farmaha","sequence":"additional","affiliation":[{"name":"Department of Pathology, Augusta University, Augusta, GA 30909, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jana","family":"Woodall","sequence":"additional","affiliation":[{"name":"Department of Pathology, Augusta University, Augusta, GA 30909, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ravindra","family":"Kolhe","sequence":"additional","affiliation":[{"name":"Department of Pathology, Augusta University, Augusta, GA 30909, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"16242","published-online":{"date-parts":[[2026,7,23]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","unstructured":"Bray F, Laversanne M, Sung H, Ferlay J, Siegel RL, Soerjomataram I, et al. 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