Department of Pathology and Laboratory Medicine, Perelman School of Medicine at the University of Pennsylvania, 3400 Spruce Street, Philadelphia, PA, 19104, USA.
Mol Diagn Ther. 2023 May;27(3):371-381. doi: 10.1007/s40291-022-00638-7. Epub 2023 Jan 23.
Determination of isocitrate dehydrogenase (IDH) 1/2 mutational status is crucial for a glioma diagnosis. It is common for IDH mutational status to be determined via a two-step algorithm that utilizes immunohistochemistry studies for IDH1 R132H, the most frequent variant, followed by next-generation sequencing studies for immunohistochemistry-negative or immunohistochemistry-equivocal cases. The objective of this study was to evaluate adding a rapid real-time polymerase chain reaction (RT-PCR) assay to the testing algorithm. METHODS: We validated a modified, commercial, qualitative, RT-PCR assay with the ability to detect 14 variants in IDH1/2 in formalin-fixed paraffin-embedded glioma tumor specimens. The assay was validated using 51 tumor formalin-fixed paraffin-embedded specimens. During clinical implementation of this assay, 48 brain tumor specimens were assessed for IDH result concordance and turnaround time to result.
Concordance between the RT-PCR and sequencing and IHC studies was 100%. This RT-PCR assay also showed concordant results with IHC for IDH1 R132H for 11 of the 12 (92%) tumor specimens with IDH mutations. The RT-PCR assay yielded faster results (average 2.6 days turnaround time) in comparison to sequencing studies (17.9 days), with complete concordance.
In summary, we report that this RT-PCR assay can reliably be performed on formalin-fixed paraffin-embedded specimens and has a faster turnaround time than sequencing assays and can be clinically implemented for determination of IDH mutation status for patients with glioma.
测定异柠檬酸脱氢酶（IDH）1/2 突变状态对于胶质瘤的诊断至关重要。通常采用两步算法来确定 IDH 突变状态，该算法首先使用免疫组织化学研究 IDH1 R132H，这是最常见的变体，然后对免疫组织化学阴性或免疫组织化学不确定的病例进行下一代测序研究。本研究的目的是评估在检测算法中添加快速实时聚合酶链反应（RT-PCR）检测。
我们验证了一种改良的、商业的、定性的 RT-PCR 检测方法，该方法能够检测福尔马林固定石蜡包埋的胶质瘤肿瘤标本中的 14 种 IDH1/2 变体。该检测方法使用 51 个肿瘤福尔马林固定石蜡包埋标本进行验证。在该检测方法的临床实施过程中，评估了 48 个脑肿瘤标本的 IDH 结果一致性和获得结果的周转时间。
RT-PCR 与测序和免疫组织化学研究之间的一致性为 100%。该 RT-PCR 检测方法还与 12 个 IDH 突变肿瘤标本中的 11 个（92%）IDH1 R132H 的免疫组织化学结果一致。与测序研究（17.9 天）相比，RT-PCR 检测方法的结果更快（平均 2.6 天的周转时间），并且具有完全的一致性。
总之，我们报告称，该 RT-PCR 检测方法可以可靠地在福尔马林固定石蜡包埋标本上进行，并且比测序检测方法具有更快的周转时间，可用于临床确定胶质细胞瘤患者的 IDH 突变状态。
