Abstract
Many diseases have their treatment options narrowed and end up being fatal if detected during later stages. As a consequence, point-of-care devices have an increasing importance for routine screening applications in the health sector due to their portability, fast analyses and decreased cost. For that purpose, a multifunctional chip was developed and tested using gold nanoprobes to perform RNA optical detection inside a microfluidic chip without the need of molecular amplification steps. As a proof-of-concept, this device was used for the rapid detection of chronic myeloid leukemia, a hemato-oncological disease that would benefit from early stage diagnostics and screening tests. The chip passively mixed target RNA from samples, gold nanoprobes and saline solution to infer a result from their final colorimetric properties. An optical fiber network was used to evaluate its transmitted spectra inside the chip. Trials provided accurate output results within 3 min, yielding signal-to-noise ratios up to 9 dB. When compared to actual state-of-art screening techniques of chronic myeloid leukemia, these results were, at microscale, at least 10 times faster than the reported detection methods for chronic myeloid leukemia. Concerning point-of-care applications, this work paves the way for other new and more complex versions of optical based genosensors.
Highlights
Nanoparticle-based bioanalyte detection has been at the forefront of miniaturized systems for molecular diagnostics at point of care (POC) testing
To demonstrate the impact of this microfluidic chip for molecular diagnostics, gold-nanoprobe (Au-nanoprobe) assays were applied in the detection of BCR-ABL1 fusion transcript (RNA), which is the molecular hallmark of chronic myeloid leukemia (CML)
Fabrication and characterization of the chip, three case studies with increasing complexity were performed and analyzed in the present report: first, Au-nanoprobes were used with and without salt to make the proof of concept; second, BCR-ABL1 synthetic oligonucleotides were combined with Au-nanoprobes to evaluate the mixing and cross-linking behavior under mixing and optical detection inside the microchannel; and third, the system performance was assessed using total RNA extracted from a CML cell line, which is the surrogate model for the disease, mimicking real clinical sample screening
Summary
Nanoparticle-based bioanalyte detection has been at the forefront of miniaturized systems for molecular diagnostics at point of care (POC) testing. Fabrication and characterization of the chip, three case studies with increasing complexity were performed and analyzed in the present report: first, Au-nanoprobes were used with and without salt to make the proof of concept; second, BCR-ABL1 synthetic oligonucleotides were combined with Au-nanoprobes to evaluate the mixing and cross-linking behavior under mixing and optical detection inside the microchannel; and third, the system performance was assessed using total RNA extracted from a CML cell line, which is the surrogate model for the disease, mimicking real clinical sample screening These results on-chip, obtained without requiring any amplification steps, were faster than the standard laboratory operation process at macroscale[7] and unprecedented at microscale even when compared with other similar and up to date methods[39,40]. These devices are suitable for the fast screenings required in medical care and POC, and key to unlock new and more complex versions of optical based genosensors, either to diagnose CML or other similar diseases
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