Mass spectrometry detection 2-HG to quickly diagnose IDH mutations

Traditional PCR sequencing and immunohistomydization require more than 1 day to get the test results, which is not suitable for rapid diagnosisTherefore, on the basis of previous discoveries, the author proposes to apply GC-MS method to carry out rapid detection of 2-HG- From the article chapter
(Ref: Xu H, et alLab Invest2019 Apr;99 (4):588-598doi: 10.1038/s41374-018-0163-zEpub 2018 Dec 20.:metastatic dehydrogenase (IDH) mutations in solid tumors, including gliomas, are specifically accumulated 2-hydroxyl diagacid (2-HG); Currently, 2-HG testing is performed by gas phase or liquid chromatography-mass spectrometry (GC-or-mass spectrometry) or biochemical, but takes a long timeHao Xu of Huashan Hospital, affiliated with Fudan University Hospital, reported that Professor Mao Ying's team at Huashan Hospital, based on the matrix-assisted laser desigenization/ionization-flight time mass spectrometry (MALDI-TOF), proposed a method for rapid detection of 2-HG in glioma to accurately determine the IDH mutation, the results were published online in December 2018 in Laboratoryresearchers first tested the idh1 mutation in 220 glioma samplesAlthough PCR sequencing and immunohislification are highly sensitive and specific, it takes 1-2 days for rapid diagnosis (Figure 1)Figure 1IDH mutation test resultssubsequently, the authors performed GC-MS tests on 58 cases of IDH1 mutant glioma identified in sequence, and found that 49 of them (84.5%) had 2-HG content slower than 2mM/gThe 2-HG content was significantly higher than low-grade gliomas in GBMCompared with wild IDH1 tumors, when the level of 2-HG is higher than 2mM/g, H3K79 dimethylation increased significantly in 6 idh1 mutant tumors (P 0.0001); Figure 2 Quantitatively detect 2-HG in idh1 mutant tumors traditional PCR sequencing and immune grouping methods take more than 1 day to get the test results, not suitable for rapid diagnosis Therefore, on the basis of previous discoveries, the author proposes to apply GC-MS method to carry out rapid detection of 2-HG The standard GC-MS method can take up to 3 hours, and only 1 hour after adjusting the sample preparation process (Figure 3) Figure 3 Method for quick detection of 2-HG 87 glioma samples, PCR sequencing identified 58 IDH1 mutations A thin mass spectrometry was detected in all 58 idh1 mutation samples None of the 29 IDH1 wild samples were detected at 2-HG (Figure 4) figure 4 The streamlined GC-MS method can accurately determine the situation of the IDH1 mutation to save time further, the author truncates the capillary in the GC instrument from a mini-pillar of 5 meters As a result, the chromatographic run time is reduced from 32 minutes to 4.2 minutes Although the results of 2-HG are affected to some extent at this time, 2-HG/Glutamate acid is still very stable and can accurately determine the IDH mutation Thus, a quick and reliable 2-HG assay can be made to identify the idh1 mutation status (Figure 5) using the mini-pillar method to sample gliomas obtained during surgery figure 5 Mini-bar method for rapid detection , the authors developed a method for rapid detection of 2-HG in surgery to diagnose gliomaIDH mutations, which have good clinical value.