Isocitrate dehydrogenase(IDH) genotype plays a significant role in precise diagnosis of glioma.An air flow assisted desorption electrospray ionization(AFADESI) mass spectrometry(MS) with rapid sample pretreatment using tissue smear instead of tissue cryo section was developed for rapid determination of the gene status of IDH in human glioma.Using intensity ratio of 2 hydroxyglutaric acid to arachidonic acid as an evaluation index,the IDH gene status of glioma tissue could be quickly identified.Quantitative evaluation as well as investigation on the feasibility for ambient mass spectrometry to analyze the metabolites confirmed that the tissue smear method was capable of rapid analysis of metabolites with high simplicity,high repeatability and high coverage.Receiver operating characteristic curve(ROC) demonstrated that the method possessed a strong diagnostic capacity for identifying the gene status of IDH in glioma.Therefore,the developed method could be expected to be a novel technique for precise diagnosis of glioma.