Development of a sensitive, specific and cost-effective T-ARMS PCR assay for the genotyping of R132H of IDH1 gene in glioma patients

The discovery of isocitrate dehydrogenase isoform 1 (IDH1) mutation as a key molecular marker has resulted in change glial tumour classification [1]. IDH1 mutations are commonly gliomas, particularly low-grade gliomas and secondary glioblastoma [2]. p.R132H (c.395G>A) accounted for more than 90% the IDH1/2 had significant association with clinical outcomes. cause gain-of-function resulting formation an oncometabolite, R-2-hydroxyglutarate instead α-ketoglutarate implying disruption oxidative decarboxylation Kreb’s cycle other cellular mechanisms [3,4,5]. Immunohistochemistry (IHC) Sanger sequencing is most common approach used diagnostics. Nevertheless, both IHC methods have shortcomings. prone to miss out on if samples poor quality, it also been reported low sensitivity when compared sequencing-based techniques [6,7].
 
 This study aimed develop sensitive, specific cost-effective assay genotyping glioma patients order shorten time required confirmatory diagnosis be made. tetra primer amplification refractory system polymerase chain reaction (T-ARMS PCR) was this validate applicability assay. A total 61 specimens were collected genomic DNA isolated from all them. All subjected endpoint PCR detection. T-ARMS developed optimized prior screening samples, comparative analysis carried out.
 Overall, found 45.90% (n=28/61) prevalent significantly associated gender, subtypes grading, location lesions (p=<0.05). With F1 score 0.966, we sensitivity, specificity accuracy 100% (95% CI: 87.94-100.00%), 93.94% 80.39-98.92%) 96.72%, respectively. To compare published studies, meta-analysis detected one case p.R132C (c.394C>T), howbeit could double (c.394C>T) or single p.R132Y (c.394_395CG>TA). However, able detect p.R132G (c.394C>G) via by whereas excluded mutation, suggesting very p.R132HH only.
 Hereby, performance sheds light that can adapted preoperative intraoperative diagnosis.