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Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:

2024 04

Page:

623-628

Research Field:

Publishing date:

Info

Title:

Development and clinical validation of a high-throughput next-generation sequencing gene panel for pan-tumor mutations detection

Author(s):

ZHANG Xiaomi¹; LIANG Zhikun²; FANG Peng²; CHEN Siyuan²; ZHOU Nan²; JIANG Xiwen¹; 2

1.Guangdong Pharmaceutical University,Guangdong Guangzhou 510006,China;2.Daan Gene Co.,Ltd.,Guangdong Guangzhou 510665,China.

Keywords:

high-throughput sequencing; gene panel; pan-tumor; genetic mutations

PACS:

R73-3

DOI:

10.3969/j.issn.1672-4992.2024.04.006

Abstract:

Objective:To develop a high-throughput sequencing gene panel for the detection of personalized pan-cancer mutation genes and validate its clinical applicability.Methods:A total of 223 tumor-associated genes were selected based on public databases and primer sets were designed for the gene panel.Laboratory validation of the analytical performance,including accuracy,repeatability,and detection limit,was conducted using GeneWell¡kformalin-fixed paraffin-embedded (FFPE) reference standards,wild-type standards,pan-cancer 800 reference standards,and Horizon Discovery¡k cfDNA reference standards.Clinical validation was performed using 10 tissue samples and 2 cfDNA samples.Results:The gene panel exhibited 100% concordance for both positive and negative detection in terms of accuracy.Repeatability was shown to be excellent using the FFPE reference standard containing 5% structural variant,while the cfDNA reference standard showed moderate repeatability.The gene panel achieved stable detection at a minimum level of 2.5%,with a minimum detectable limit of 1%.The lowest detectable limit for the cfDNA reference standard was 0.25%.The gene panel demonstrated good consistency with the control method when applied to clinical samples.Conclusion:In this study,we successfully developed a high-throughput gene panel for 223 tumor-associated genes and validated its analytical performance and clinical applicability.This method has been proven to have the potential to assist clinical diagnosis and treatment.

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