Microfluidics for rapid detection of isocitrate dehydrogenase 1 mutation for intraoperative application

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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.

ABBREVIATIONSAUC = area under the curve; FFPE = formalin-fixed paraffin-embedded; IDH1 = isocitrate dehydrogenase 1; IHC = immunohistochemistry; PCR = polymerase chain reaction; ROC = receiver operating characteristic.
Article Information

Contributor Notes

Correspondence Jin-song Wu, Glioma Surgery Division, Neurological Surgery Department, Huashan Hospital, Shanghai Medical College, Fudan University, 12 Middle Wulumuqi Rd., Shanghai 200040, China, email: wjsongc@126.com.* Drs. Aibaidula and Zhao contributed equally to this work.INCLUDE WHEN CITING Published online November 6, 2015; DOI: 10.3171/2015.4.JNS141833.Disclosure The authors report no conflict of interest concerning the materials or methods used in this study or the findings specified in this paper.

© AANS, except where prohibited by US copyright law.

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