A targeted next-generation sequencing panel for identification of clinically relevant mutation profiles in solid tumours

Abstract Targeted next generation sequencing (NGS) using multigene panels has become an effective tool for comprehensive genomic analysis in cancer, overcoming limitations of single gene assays. Nonetheless, outsourcing these assays to external laboratories and the extended turnaround time (~ 3 week...

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Main Authors: Kakoli Das, Mandy Li Ian Tay, Elena Yaqing Yong, Khoon Leong Chuah
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-08039-6
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author Kakoli Das
Mandy Li Ian Tay
Elena Yaqing Yong
Khoon Leong Chuah
author_facet Kakoli Das
Mandy Li Ian Tay
Elena Yaqing Yong
Khoon Leong Chuah
author_sort Kakoli Das
collection DOAJ
description Abstract Targeted next generation sequencing (NGS) using multigene panels has become an effective tool for comprehensive genomic analysis in cancer, overcoming limitations of single gene assays. Nonetheless, outsourcing these assays to external laboratories and the extended turnaround time (~ 3 weeks) required for obtaining results may impede timely clinical management of cancer patients. We developed an oncopanel targeting 61 cancer-associated genes and validated its efficacy by performing NGS on 43 unique samples including clinical tissues, external quality assessment samples, and reference controls. The assay detected 794 mutations including all 92 known variants from orthogonal methods. Overall performance measures of the assay showed 99.99% repeatability and 99.98% reproducibility. Likewise, sensitivity to detect unique variants was 98.23%, with specificity at 99.99%, precision at 97.14% and accuracy at 99.99% all at 95% CI. Notably, clinically actionable mutations were observed in key genes such as KRAS, EGFR, ERBB2, PIK3CA, TP53 and BRCA1. The average turnaround time from sample processing to results was reduced to 4 days. These findings demonstrate a sensitive, high throughput oncopanel that is suitable for use in routine clinical testing. The shorter turnaround time of the assay has the potential to significantly improve patient care by facilitating more timely and personalized clinical interventions.
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issn 2045-2322
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spelling doaj-art-9d906ecec38147d58cfdd72f46b033102025-08-20T03:37:30ZengNature PortfolioScientific Reports2045-23222025-07-0115111110.1038/s41598-025-08039-6A targeted next-generation sequencing panel for identification of clinically relevant mutation profiles in solid tumoursKakoli Das0Mandy Li Ian Tay1Elena Yaqing Yong2Khoon Leong Chuah3Department of Pathology, Tan Tock Seng HospitalDepartment of Pathology, Tan Tock Seng HospitalDepartment of Pathology, Tan Tock Seng HospitalDepartment of Pathology, Tan Tock Seng HospitalAbstract Targeted next generation sequencing (NGS) using multigene panels has become an effective tool for comprehensive genomic analysis in cancer, overcoming limitations of single gene assays. Nonetheless, outsourcing these assays to external laboratories and the extended turnaround time (~ 3 weeks) required for obtaining results may impede timely clinical management of cancer patients. We developed an oncopanel targeting 61 cancer-associated genes and validated its efficacy by performing NGS on 43 unique samples including clinical tissues, external quality assessment samples, and reference controls. The assay detected 794 mutations including all 92 known variants from orthogonal methods. Overall performance measures of the assay showed 99.99% repeatability and 99.98% reproducibility. Likewise, sensitivity to detect unique variants was 98.23%, with specificity at 99.99%, precision at 97.14% and accuracy at 99.99% all at 95% CI. Notably, clinically actionable mutations were observed in key genes such as KRAS, EGFR, ERBB2, PIK3CA, TP53 and BRCA1. The average turnaround time from sample processing to results was reduced to 4 days. These findings demonstrate a sensitive, high throughput oncopanel that is suitable for use in routine clinical testing. The shorter turnaround time of the assay has the potential to significantly improve patient care by facilitating more timely and personalized clinical interventions.https://doi.org/10.1038/s41598-025-08039-6Targeted sequencingCancerGenomicsSomatic mutationOncopanelNGS
spellingShingle Kakoli Das
Mandy Li Ian Tay
Elena Yaqing Yong
Khoon Leong Chuah
A targeted next-generation sequencing panel for identification of clinically relevant mutation profiles in solid tumours
Scientific Reports
Targeted sequencing
Cancer
Genomics
Somatic mutation
Oncopanel
NGS
title A targeted next-generation sequencing panel for identification of clinically relevant mutation profiles in solid tumours
title_full A targeted next-generation sequencing panel for identification of clinically relevant mutation profiles in solid tumours
title_fullStr A targeted next-generation sequencing panel for identification of clinically relevant mutation profiles in solid tumours
title_full_unstemmed A targeted next-generation sequencing panel for identification of clinically relevant mutation profiles in solid tumours
title_short A targeted next-generation sequencing panel for identification of clinically relevant mutation profiles in solid tumours
title_sort targeted next generation sequencing panel for identification of clinically relevant mutation profiles in solid tumours
topic Targeted sequencing
Cancer
Genomics
Somatic mutation
Oncopanel
NGS
url https://doi.org/10.1038/s41598-025-08039-6
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