title: Microfluidics for rapid detection of isocitrate dehydrogenase 1 mutation for intraoperative application.
abstract: OBJECT Conventional methods for isocitrate dehydrogenase 1 (IDH1) detection, such as DNA sequencing and immunohistochemistry, are time- and labor-consuming and cannot be applied for intraoperative analysis. To develop a new approach for rapid analysis of IDH1 mutation from tiny tumor samples, this study used microfluidics as a method for IDH1 mutation detection. METHODS Forty-seven glioma tumor samples were used; IDH1 mutation status was investigated by immunohistochemistry and DNA sequencing. The microfluidic device was fabricated from polydimethylsiloxane following standard soft lithography. The immunoanalysis was conducted in the microfluidic chip. Fluorescence images of the on-chip microcolumn taken by the charge-coupled device camera were collected as the analytical results readout. Fluorescence signals were analyzed by NIS-Elements software to gather detailed information about the IDH1 concentration in the tissue samples. RESULTS DNA sequencing identified IDH1 R132H mutation in 33 of 47 tumor samples. The fluorescence signal for IDH1-mutant samples was 5.49 ± 1.87 compared with 3.90 ± 1.33 for wild type (p = 0.005). Thus, microfluidics was capable of distinguishing IDH1-mutant tumor samples from wild-type samples. When the cutoff value was 4.11, the sensitivity of microfluidics was 87.9% and the specificity was 64.3%. CONCLUSIONS This new approach was capable of analyzing IDH1 mutation status of tiny tissue samples within 30 minutes using intraoperative microsampling. This approach might also be applied for rapid pathological diagnosis of diffuse gliomas, thus guiding personalized resection.
authors.affiliation: Glioma Surgery Division, Neurological Surgery Department, Huashan Hospital.
authors.forename: Abudumijiti
authors.initials: A
authors.lastname: Aibaidula
authors.affiliation: Department of Environmental Science and Engineering, Institute of Biomedical Sciences, and.
authors.forename: Wang
authors.initials: W
authors.lastname: Zhao
authors.affiliation: Glioma Surgery Division, Neurological Surgery Department, Huashan Hospital.
authors.forename: Jin-Song
authors.initials: JS
authors.lastname: Wu
authors.affiliation: Department of Neuropathology, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
authors.affiliation: Glioma Surgery Division, Neurological Surgery Department, Huashan Hospital.
authors.forename: Zhi-Feng
authors.initials: ZF
authors.lastname: Shi
authors.affiliation: Department of Environmental Science and Engineering, Institute of Biomedical Sciences, and.
authors.forename: Lu-Lu
authors.initials: LL
authors.lastname: Zheng
authors.affiliation: Glioma Surgery Division, Neurological Surgery Department, Huashan Hospital.
authors.forename: Ying
authors.initials: Y
authors.lastname: Mao
authors.affiliation: Glioma Surgery Division, Neurological Surgery Department, Huashan Hospital.
authors.forename: Liang-Fu
authors.initials: LF
authors.lastname: Zhou
authors.affiliation: Department of Environmental Science and Engineering, Institute of Biomedical Sciences, and.
authors.forename: Guo-Dong
authors.initials: GD
authors.lastname: Sui
mesh_terms: D000328:Adult; D019540:Area Under Curve; D001921:Brain / Q000378:metabolism; D001932:Brain Neoplasms / Q000235:genetics / Q000378:metabolism / Q000601:surgery; D004252:DNA Mutational Analysis / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D005240:Feasibility Studies; D005260:Female; D005910:Glioma / Q000235:genetics / Q000378:metabolism / Q000601:surgery; D006801:Humans; D007150:Immunohistochemistry; D007521:Isocitrate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D008297:Male; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D009154:Mutation; D019635:Neurosurgical Procedures / Q000379:methods; D011446:Prospective Studies; D012372:ROC Curve; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D007521:Isocitrate Dehydrogenase; C543588:IDH1 protein, human
keywords: AUC = area under the curve; FFPE = formalin-fixed paraffin-embedded; IDH1 = isocitrate dehydrogenase 1; IHC = immunohistochemistry; PCR = polymerase chain reaction; ROC = receiver operating characteristic; glioma; isocitrate dehydrogenase 1; microfluidics; neurosurgery; oncology