Prospects of Microfluidic Technology in Nucleic Acid Detection Approaches

Abstract

Conventional diagnostic techniques are based on the utilization of analyte sampling, sensing and signaling on separate platforms for detection purposes, which must be integrated to a single step procedure in point of care (POC) testing devices. Due to the expeditious nature of microfluidic platforms, the trend has been shifted toward the implementation of these systems for the detection of analytes in biochemical, clinical and food technology. Microfluidic systems molded with substances such as polymers or glass offer the specific and sensitive detection of infectious and noninfectious diseases by providing innumerable benefits, including less cost, good biological affinity, strong capillary action and simple process of fabrication. In the case of nanosensors for nucleic acid detection, some challenges need to be addressed, such as cellular lysis, isolation and amplification of nucleic acid before its detection. To avoid the utilization of laborious steps for executing these processes, advances have been deployed in this perspective for on-chip sample preparation, amplification and detection by the introduction of an emerging field of modular microfluidics that has multiple advantages over integrated microfluidics. This review emphasizes the significance of microfluidic technology for the nucleic acid detection of infectious and non-infectious diseases. The implementation of isothermal amplification in conjunction with the lateral flow assay greatly increases the binding efficiency of nanoparticles and biomolecules and improves the limit of detection and sensitivity. Most importantly, the deployment of paper-based material made of cellulose reduces the overall cost. Microfluidic technology in nucleic acid testing has been discussed by explicating its applications in different fields. Next-generation diagnostic methods can be improved by using CRISPR/Cas technology in microfluidic systems. This review concludes with the comparison and future prospects of various microfluidic systems, detection methods and plasma separation techniques used in microfluidic devices.

Recent Uses of Paper Microfluidics in Isothermal Nucleic Acid Amplification Tests

“…One review paper summarized the same characteristics of various isothermal amplification methods. In summary, RPA has the shortest reaction time between 5-7 min and only uses one primer [ 149 ]. The quick reaction time and simplicity of only having one primer could be one reason why RPA is a popular choice for paper-based amplification.…”

“…Other comparisons of isothermal methods recognize that HDA and NASBA also average the same time of approximately 1.5 h for nucleic acid amplification and use only one pair of primers for these isothermal methods, but NASBA can be utilized with a larger variety of templates. According to this review paper [ 149 ], NASBA targets can include ssDNA, transfer-messenger RNA, and ribosomal RNA, making it an amplification method worth further exploration.…”

“…Such diversity can also be seen with LAMP, RCA, SDA, and EXPAR to lesser extents, while RAA, MIRA, NASBA, and HDA are almost exclusively used with colorimetric detection. Another review paper summarized the advantages and disadvantages of common detection methods [ 149 ]. The most notable advantages were low cost, high signal sensitivity, and simplicity across multiple detection methods (electrochemical, colorimetric, chemiluminescence, and fluorescence).…”

The development of droplet-based microfluidic virus detection technology for human infectious diseases

“…Detection based on microfluidics has the advantages of low sample consumption, fast detection speed, simple operation, multifunctional integration, small size, and portability. 9 Therefore, it is particularly suitable for developing point-of-care (POC) diagnostics and is considered to have enormous potential for the development and wide application prospects of virus detection technology research. 7,9…”

“…9 Therefore, it is particularly suitable for developing point-of-care (POC) diagnostics and is considered to have enormous potential for the development and wide application prospects of virus detection technology research. 7,9…”