Microfluidic device for novel breast cancer screening by blood test using miRNA beacon probe

Abstract

Breast cancer is identified as the highest cause of death in women suffering from cancer. Early diagnosis is the key to increase the survival of breast cancer victims. Molecular diagnosis using biomarkers have advanced much in the recent years. The cost involved in such diagnosis is not affordable for most of the population. The concept being investigated here is to realize a simple diagnosis system for screening cancer by way of a blood test utilizing a miRNA based biomarker with a complementary molecular beacon probe. A microfluidic platform was designed and attached with a fluorescence reader, which is portable and cost effective. Experiments were performed with 51 blood samples of which 30 were healthy and 21 were positive for breast cancer, collected against institutional human ethical clearance, IHEC 16/180-7-9-2016. miRNA 21 was chosen as the biomarker because it is overexpressed 4-fold in the serum of breast cancer patients. This work involved design of an experiment to prove the concept of miRNA over expression followed by detection of miRNA 21 using the microfluidic platform attached with a fluorescence reader and validation of the results using quantitative Real Time Polymerase Chain Reaction (qRT-PCR). The results obtained from the microfluidic device concurred with qRT-PCR results. The device is suitable for point-of-care application in a mass-screening programme. The study also has revealed that the stage of the cancer could be indicated by this test, which will be further useful for deciding a therapeutic regime.

Perspectives on the Role of Histone Modification in Breast Cancer Progression and the Advanced Technological Tools to Study Epigenetic Determinants of Metastasis

“…Point of care (POC) systems were also developed in recent years. A miRNA-based microfluidic POC platform integrated with fluorescence reader was demonstrated to screen and detect miR-21 in the blood samples of breast cancer patients within 30 min (Salim et al, 2017). A further step in this direction was taken by using a power-free microfluidic chip to detect miR-196a and miR-331 (Kim et al, 2019).…”

Microfluidic and Paper-Based Devices for Disease Detection and Diagnostic Research

“…These devices are different from LFSAs and µPADS because PDMS microfluidic devices often rely on an external pressure source to induce the flow of fluids through the device instead of diffusion-driven wicking of fluids across a piece of paper. Microfluidic techniques to detect specific biomolecules for disease identification and progression include the use of microfluidic trapping arrays [ 42 ], droplet encapsulation of samples [ 42 , 43 ], on-chip genetic amplification [ 38 ], flow-based filtration of samples [ 39 ], and capturing biomolecules using antibodies [ 39 , 40 ].…”

“…PDMS microfluidic devices have been developed to control the mixing of samples with reagents through complex geometries of channels and arrays to produce a measurable output [ 38 , 39 , 40 , 41 ]. Other microfluidic devices incorporate droplet generation and trapping to detect biomolecules [ 42 , 43 ].…”

“…Other microfluidic devices incorporate droplet generation and trapping to detect biomolecules [ 42 , 43 ]. Analytical outputs for these platforms include fluorescent microscopy [ 38 , 40 , 43 ], spectroscopy [ 39 ], color production and intensity [ 41 ], and analysis of individual droplets [ 42 , 43 ]. A device developed by Salim et al was used to perform genetic amplification on-chip.…”

“…A device developed by Salim et al was used to perform genetic amplification on-chip. The microfluidic droplet generator was used to perform quantitative reverse transcription polymerase chain reaction (qRT-PCR) in order to detect overexpression of miRNA 21 as a biomarker for breast cancer in patients by measuring a fluorescent output [ 38 ]. Other microfluidic devices utilized antibodies immobilized within the flow channels to capture desired proteins [ 39 , 40 ].…”