Towards Multiplex Molecular Diagnosis—A Review of Microfluidic Genomics Technologies

Abstract

Highly sensitive and specific pathogen diagnosis is essential for correct and timely treatment of infectious diseases, especially virulent strains, in people. Point-of-care pathogen diagnosis can be a tremendous help in managing disease outbreaks as well as in routine healthcare settings. Infectious pathogens can be identified with high specificity using molecular methods. A plethora of microfluidic innovations in recent years have now made it increasingly feasible to develop portable, robust, accurate, and sensitive genomic diagnostic devices for deployment at the point of care. However, improving processing time, multiplexed detection, sensitivity and limit of detection, specificity, and ease of deployment in resource-limited settings are ongoing challenges. This review outlines recent techniques in microfluidic genomic diagnosis and devices with a focus on integrating them into a lab on a chip that will lead towards the development of multiplexed point-of-care devices of high sensitivity and specificity.

Semi-Automated Microfluidic Device Combined with a MiniPCR-Duplex Lateral Flow Dipstick for Screening and Visual Species Identification of Lymphatic Filariae

“…The multiplex detection of more than one species of the pathogen on a single strip offers the advantages of increased speed and lower cost. Multiplex rapid lateral flow diagnostics offers the potential of distinguishing among multiple pathogens, thereby facilitating diagnosis and improving patient care [8,10,18].…”

“…The processing capacity of laboratories is limited because highly trained personnel must supervise the analysis and the biological sample must be transported from the patient at the point of collection to the lab and the results are usually obtained ≥24 h later [7]. Furthermore, laboratory-based diagnosis cannot meet the demands of mass identification of pathogens [8][9][10].…”

“…Microfluidic platforms require and consume only minute amounts of samples and reagents, which minimizes waste and expense, and they offer the unique physical advantage of microscale fluid flow [11]. Innovations in microfluidic technologies have facilitated the development of portable, robust, accurate, and sensitive diagnostic devices for point-of-care use [10]. A microfluidic chip, which is often referred to as a lab on a chip, is a chip that was developed for use in life science and biology that is characterized by fluid flow along micro-scale channels (i.e., microfluidics) [12].…”

Dengue Detection: Advances in Diagnostic Tools from Conventional Technology to Point of Care

“…LOC embodies numerous steps into one platform, which allows real-time detection while only using a small amount of the sample [174]. These steps include biochemical reactions, transportation, product detection, and sample/analyte concentrators [175]. There are several LOC technologies that have been developed to detect various viral targets, as well as devices that incorporate micro/nanoparticles [135,[176][177][178].…”

An Overview on Microfluidic Systems for Nucleic Acids Extraction from Human Raw Samples

“…before proceeding with membrane lysis. Membrane lysis methods can be categorized as: (1) mechanical—by grinding, freeze-thawing, potterization, centrifugation, sonication or bead-beating; (2) chemical—by detergents, solubilizing membrane lipids, or by chaotropic agents, perforating cell membranes with the denaturation of trans-membrane proteins, by enzymatic digestion, or by osmotic shock; (3) thermal—heating treatments; or (4) electrical (by membrane dielectric breakdown) [ 14 , 87 , 88 ]. Sample lysis can be accomplished by a single or a combined use of the above-mentioned methods, according to the type of cell to treat and, eventually, according to the needed quality or yields of the extraction.…”