A method of detecting IDH2 gene mutations in angioimmunoblastic T-cell lymphoma based on droplet digital PCR
10.3760/cma.j.cn121090-20241016-00396
- VernacularTitle:基于微滴式数字PCR建立血管免疫母细胞性T细胞淋巴瘤IDH2基因突变检测方法
- Author:
Xi HUANG
1
;
Yi MIAO
;
Xiao XIAO
;
Jing LI
;
Hui JIN
;
Jianyong LI
;
Wenyu SHI
Author Information
1. 南通大学附属医院肿瘤科、血液科，南通 226001
- Publication Type:Journal Article
- Keywords:
Lymphoma, T-cell, peripheral;
Lymphoma, T-cell, angioimmunoblastic;
IDH2;
Mutation detection, Droplet digital PCR
- From:
Chinese Journal of Hematology
2025;46(6):544-550
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a method to detect IDH2 gene mutations in the cell-free DNA (cfDNA) sample of angioimmunoblastic T-cell lymphoma (AITL) based on the digital droplet PCR (ddPCR) method. The performance of this method was evaluated.Methods:Specific primers and probes were designed for IDH2 p. R172K, IDH2 p. R172M, and IDH2 p. R172W mutations, and primer combinations were optimized to establish the optimal ddPCR reaction system. In addition, the accuracy, precision, specificity, and detection limit of the method were evaluated using clinical cfDNA samples obtained from 40 patients with AITL at the First Affiliated Hospital of Nanjing Medical University.Results:The IDH2 R172K/M/W F2/R2 primer combination was selected to establish the ddPCR detection method for IDH2 R172K/M/W mutations. Specific standard samples showed no false-positive results. The accuracy test revealed that the relative deviation of the standard sample measurements was within ±10%. The detection limit test indicated that the method could detect 1 ng/μl with a 0.2% mutation rate for IDH2 R172K/M/W mutations. The precision measurements (inter-experimenter, inter-run, and inter-laboratory) all had a coefficient of variation of <5%. ddPCR was applied to 40 clinical samples with IDH2 R172K/M/W mutations, and the results were fully consistent with gene sequencing.Conclusion:ddPCR can detect IDH2 gene mutations in cfDNA from AITL with high sensitivity, specificity, and absolute quantification. This method offers a simple, rapid, and reliable approach for diagnosing and monitoring.
