METHODS: In the current study, IDH1-R132H mutation was analyzed in tumor tissue with paired cell-free DNA (cfDNA) in patients with ADG (n = 45) using IHC and competitive allele-specific Taqman PCR (CAST-PCR). Genomic DNA was extracted from formalin-fixed paraffin-embedded (FFPE) tissue and matched serum for cfDNA using commercially available kits. CAST-PCR with IHC for the detection of IDH1-R132H mutation was also compared.
RESULTS: The IDH1-R132H mutation was detected in 46.67% (21/45) cases and 57.78% (26/45) cases using IHC and allele-specific CAST-PCR. In cfDNA of matched IDH1-mutant FFPE tissue DNA, IDH1-R132H mutation was detected in 11.54% (3/26) using CAST-PCR. The concordance rate for IDH1-R132Hmutation between IHC and CAST-PCR was 80.77% (21/26).
CONCLUSIONS: The CAST-PCR assay is more precise and sensitive for IDH1-R132Hdetection than traditional IHC, and IDH1-R132H mutation detection using cfDNA may add to the current methods of glioma genomic characterization.
方法:在目前的研究中,使用IHC和竞争性等位基因特异性TaqmanPCR(CAST-PCR)分析ADG患者(n=45)中具有配对无细胞DNA(cfDNA)的肿瘤组织中的IDH1-R132H突变。使用可商购的试剂盒从福尔马林固定的石蜡包埋(FFPE)组织中提取基因组DNA并匹配cfDNA的血清。还比较了用IHC检测IDH1-R132H突变的CAST-PCR。
结果:使用IHC和等位基因特异性CAST-PCR,在46.67%(21/45)例和57.78%(26/45)例中检测到IDH1-R132H突变。在匹配的IDH1突变体FFPE组织DNA的cfDNA中,使用CAST-PCR检测到IDH1-R132H突变11.54%(3/26)。IHC与CAST-PCR的IDH1-R132H突变符合率为80.77%(21/26)。
结论:与传统的IHC相比,CAST-PCR对IDH1-R132H检测更精确和灵敏,使用cfDNA检测IDH1-R132H突变可能会增加目前的神经胶质瘤基因组表征方法。