A portable microfluidic platform for rapid molecular diagnostic testing of patients with myeloproliferative neoplasms

Abstract

The ability to simultaneously detect JAK2 V617F and MPL W515K/L mutations would substantially improve the early diagnosis of myeloproliferative neoplasms (MPNs) and decrease the risk of arterial thrombosis. The goal of this study is to achieve a point of care testing platform for simultaneous analysis of major genetic alterations in MPN. Here, we report a microfluidic platform including a glass capillary containing polypropylene matrix that extracts genomic DNA from a drop of whole blood, a microchip for simultaneous multi-gene mutation screening, and a handheld battery-powered heating device. The µmLchip system was successfully used for point-of-care identification of the JAK2 V617F and MPL W515K/L mutations. The µmLchip assays were then validated by mutation analysis with samples from 100 MPN patients who had previously been analyzed via unlabeled probe melting curve analysis or real-time PCR. The results from the µmLchip were in perfect agreement with those from the other methods, except for one discrepant result that was negative in the unlabeled probe melting curve analysis but positive in the µmLchip. After T-A cloning, sequences of cloned PCR products revealed JAK2 V617F mutation in the sample. The portable microfluidic platform may be very attractive in developing point-of-care diagnostics for MPL W515K/L and JAK2 V617F mutations.

Development of Rapid Pharmacogenomic Testing Assay in a Mobile Molecular Biology Laboratory (2MoBiL)

“…The ability and accuracy in detecting genomic variants by using these devices have contributed to the diagnosis of hemochromatosis disease (Macleod et al, 2016) and myeloproliferative neoplasms by detecting the human p.C282Y hemochromatosis variant using the electrochemical microarray method and JAK2 p.V617F and MPL p.W515K/L variants using the loop-mediated isothermal amplification technique, respectively (Wang et al, 2017).…”

RNA-sequencing reveals that STRN, ZNF484 and WNK1 add to the value of mitochondrial MT-COI and COX10 as markers of unstable coronary artery disease

“…Especially the use of mcrochips will lead to rapid and sensitive detection of multiple targets possibly within 20 minutes [40, 41]. Portable microfluidic platforms may be very attractive in developing point-of-care diagnostics and precision medicine [42]. In additon, several machine-learning algorithms such as random forest, logistic regression, gradient boosting, and neural networks are now available to improve risk prediction [43–45].…”

Microfluidics for Biotechnology: Bridging Gaps to Foster Microfluidic Applications

“…Though this research field has developed over the past 30 years immensely, this aim is not fully realised yet. Nevertheless, the small dimensions of the devices allow, to some extent, experiments at varying locations of interest, e.g., for point of care and point of use testing (Wang et al, 2017(Wang et al, , 2019.…”