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

Abstract

S equence analysis of glioblastoma found a mutation in isocitrate dehydrogenase 1 (IDH1), which mostly resulted in the replacement of wild-type arginine in position 132 with histidine (R132H). 25 Studies have reported that the IDH1 mutation is frequently detected in diffuse astrocytoma (59%-88%), anaplastic astrocytoma (52%-78%), oligodendroglioma (68%-82%), anaplastic oligodendroglioma (60%-75%), oligoastrocytoma (50%-94%), anaplastic oligoastrocytoma (66.1%-78%), and secondary glioblastomas (50%-88%), but rarely in primary glioblastomas (3%-7%).1,14 IDH1 mutation was also closely associated with young age and better clinical outcome. 34 Currently, the detection of IDH1 mutation status can be achieved by DNA-based methods, such as DNA pyrosequencing, 10 polymerase chain reaction (PCR), 13,24 and immunohistochemistry (IHC). 5 The majority of laboratories use IHC for initial screening for IDH1 mutation and, if results are negative, use DNA sequencing for less common IDH1 mutant types. DNA sequencing provides direct abbreviatioNs AUC = area under the curve; FFPE = formalin-fixed paraffin-embedded; IDH1 = isocitrate dehydrogenase 1; IHC = immunohistochemistry; PCR = polymerase chain reaction; ROC = receiver operating characteristic. obJective 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.

Rapid detection of isocitrate dehydrogenase 1 mutation status in glioma based on Crispr-Cas12a

“…High-resolution melting is easy, fast and sensible, while it requires high quality for RT-PCR amplifier and operator 22 . There are also many detection methods for targeting 2-HG, such as the downstream metabolites of IDH-mut, including magnetic resonance spectroscopy (MRS) 23 , multiple mass spectrometry (MS) [24][25][26] and microfluidics 27 . Among them, MS is relatively mature and can quickly and accurately detect IDH mutation in surgery, which is relying on expensive mass spectrometers.…”

MinION rapid sequencing: Review of potential applications in neurosurgery

“…Recent reports on intervertebral disc disease suggest that up to 75% of the underlying etiology is attributed to genes. [ 44 ] Other diseases of neurosurgical importance found to have a component of genetic etiology include gliomas,[ 9 48 ] schwannomas,[ 24 ] meningiomas,[ 12 ] intracranial aneurysms,[ 26 38 72 73 ] arteriovenous malformations,[ 19 ] cavernous malformations,[ 21 ] subarachnoid hemorrhage,[ 25 ] idiopathic scoliosis,[ 22 ] moya-moya disease,[ 27 ] hemorrhagic, and ischemic strokes. The results generated from these genome wide association (GWA) studies, revealing specific SNPs and their association with disease, are the stepping stones for better understanding of disease risk, prediction, treatment, and prognosis.…”

Pediatric Glioblastoma

“…Numerous studies have reiterated the utility of maximal tumor removal on both progression free as well as overall survival (OS) in p-GBM (4, 9, 10, 13, 14, 51). The Children' s Cancer Group (CCG) study-945 showed that children with HGG who underwent a surgical resection of 90% or greater had a progression-free survival (PFS) of 35 ± 7% as compared with a 5-year PFS of 17 ± 4% in patients who did not (50,51). Reporting on probably the largest single-center experience of p-GBM, we have also shown that the extent of tumor excision was a strong predictor of long progression-free survival as well as OS (4).…”

“…These advanced techniques include intraoperative neuronavigation, intraoperative ultrasound, intraoperative MRI, intraoperative cortical mapping, etc. Recent technological advances utilizing microfluidic chips allow for rapid analysis of the operative specimen for molecular signatures like IDH mutation within no time (51,52). Therefore, it is possible now to make a molecular diagnosis even intraoperatively.…”