Development and characterization of reference materials for EGFR, KRAS, NRAS, BRAF, PIK3CA, ALK, and MET genetic testing

Article type: Research Article

Authors: Zhang, Wenxin^{a; 1} | Qu, Shoufang^{a; 1} | Chen, Qiong^{b; c; 1} | Yang, Xuexi^b | Yu, Jing^d | Zeng, Shuang^d | Chu, Yuxing^e | Zou, Hao^f | Zhang, Zhihong^g | Wang, Xiaowen^h | Jing, Ruilin^h | Wu, Yingsong^b | Liu, Zhipengⁱ | Xu, Ren^j | Wu, Chunyan^k | Huang, Chuanfeng^a | Huang, Jie^{a; *}

Affiliations: [a] Department of In Vitro Diagnostic Reagent, National Institutes for Food and Drug Control (NIFDC), Beijing, China | [b] Institute of Antibody Engineering, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, Guangdong, China | [c] Medical Research Center, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, China | [d] BGI Genomics, BGI-Shenzhen, Shenzhen, Guangdong, China | [e] Geneplus-Beijing Clinical Laboratory Co., Ltd., Beijing, China | [f] Novogene (Tianjin) Bioinformatics Technology Co., Ltd., Tianjin, China | [g] Guangzhou Burning Rock Dx Co., Ltd., Guangzhou, Guangdong, China | [h] Annoroad Gene Technology, Beijing, China | [i] Research Institute, Guangzhou Darui Biotechnology Co., Ltd., Guangzhou, Guangdong, China | [j] Shanghai Yuanqi Bio-Pharmaceutical Co., Ltd., Shanghai, China | [k] Department of Pathology, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China

Correspondence: [*] Corresponding author: Jie Huang, Department of In Vitro Diagnostic Reagent, National Institutes for Food and Drug Control (NIFDC), No. 2 Tiantan Xili Dongcheng District, Beijing 10050, China. E-mail: [email protected].

Note: [1] Wenxin Zhang, Shoufang Qu and Qiong Chen contributed equally to this work.

Abstract: BACKGROUND: Along with the dramatic development of molecular diagnostic testing for the detection of oncogene variations, reference materials (RMs) have become increasingly important in performance evaluation of genetic testing. OBJECTIVE: In this study, we built a set of RMs for genetic testing based on next-generation sequencing (NGS). METHOD: Solid tumor tissues were selected as the samples of RMs for preparation. NGS was used to determine and validate the variants and the mutation frequency in DNA samples. Digital PCR was used to determine the copy numbers of RNA samples. The performance of the RMs was validated by six laboratories. RESULTS: Thirty common genetic alterations were designed based on these RMs. RMs consisted of a positive reference, a limit of detection reference, and a negative reference. The validation results confirmed the performance of the RMs. CONCLUSION: These RMs may be an attractive tool for the development, validation, and quality monitoring of molecular genetic testing.

Keywords: Reference materials, genetic testing, quality control, oncogene variants

DOI: 10.3233/THC-220102

Journal: Technology and Health Care, vol. 31, no. 2, pp. 485-495, 2023