Abstract

Nucleic acid extraction (NAE) plays a vital role in molecular biology as the primary step for many downstream applications. Many modifications have been introduced to the original 1869 method. Modern processes are categorized into chemical or mechanical, each with peculiarities that influence their use, especially in point-of-care diagnostics (POC-Dx). POC-Dx is a new approach aiming to replace sophisticated analytical machinery with microanalytical systems, able to be used near the patient, at the point of care or point of need. Although notable efforts have been made, a simple and effective extraction method is still a major challenge for widespread use of POC-Dx. In this review, we dissected the working principle of each of the most common NAE methods, overviewing their advantages and disadvantages, as well their potential for integration in POC-Dx systems. At present, it seems difficult, if not impossible, to establish a procedure which can be universally applied to POC-Dx. We also discuss the effects of the NAE chemicals upon the main plastic polymers used to mass produce POC-Dx systems. We end our review discussing the limitations and challenges that should guide the quest for an efficient extraction method that can be integrated in a POC-Dx system.

Highlights

  • Nucleic acid extraction (NAE) is one of the most pivotal steps in molecular biology, being routinely used in many areas of the biological and medical sciences, as this procedure marks a starting point in any molecular diagnostic kit [1]

  • Some chemicals have the potential to affect polymer’s color, surface appearance, flexibility, and mass, generating extractable/leachable products that must be evaluated. These changes can happen due to several physicochemical reactions, such as (i) chemical interaction with polymer chain which can disturb their structure and result in depolymerization; (ii) physical interaction, that is, adsorption of chemicals into the plastics, which results in swelling and softening; or (iii) stress-associated cracking may happen due to the stresscracking agents, such as plasticizers, or adhesives used during the manufacturing of polymer parts, or even detergents or oils used during the molecular biology processes [98]

  • After almost 150 years after the first successful isolation of DNA by Friedrich Miescher, nucleic acids are central to obtaining biological information in areas as distinct as specimens’ identification for conservational purposes to the realms of personalized medicine and pharmacogenomics

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Summary

Introduction

Nucleic acid extraction (NAE) is one of the most pivotal steps in molecular biology, being routinely used in many areas of the biological and medical sciences, as this procedure marks a starting point in any molecular diagnostic kit [1]. This crucial procedure has been known for over a century and has developed substantially over the last decades. Despite the structural differences, the most commonly used methods described in the present text can be applied to DNA in its many organizational formats (chromosomal, plasmid, etc.), as well as RNA and its multidimensional formats (mRNA, rRNA, tRNA, miRNA, etc.) with minor modifications [1, 4, 5].

Method Chemical
Chemically Driven Methods
Method
Solid-Phase Nucleic Acid Extraction
Devices Used in Extraction Methods
Limitations for Implementation of Extraction Protocols in Portable Devices
Challenges for Implementation in POC Diagnostic Tests
Findings
Conclusion
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