Abstract

ADVANCEMENTS IN PCR AND ISOTHERMAL AMPLIFICATION TECHNOLOGIES

Ali Hassan Alhussain*, Waseem Ali Alquwayi, Yasser Abdrab alameer Alkuwaiti, Ahmed Mohammed Almehainy

ABSTRACT

Background: Advancements in nucleic acid amplification technologies have revolutionized molecular diagnostics. Polymerase Chain Reaction (PCR) remains a cornerstone for sensitive and specific DNA and RNA detection. However, its reliance on thermal cycling limits portability and accessibility. Isothermal amplification technologies, such as Loop-Mediated Isothermal Amplification (LAMP) and Recombinase Polymerase Amplification (RPA), provide innovative solutions, enabling rapid and robust amplification without thermal cycling. These technologies have significant implications for point-of-care (POC) testing, infectious disease diagnostics, and genomic medicine. Aim: This paper aims to review recent advancements in PCR and isothermal amplification technologies, highlighting their technical improvements, comparative advantages, and applications in clinical diagnostics, field testing, and genomic research. Methods: A systematic review of peer-reviewed literature from 2015 to 2018 was conducted, focusing on innovations in amplification technologies. Key areas of analysis included speed, sensitivity, specificity, and integration with diagnostic platforms. Results: Recent innovations in PCR include ultra-fast cycling methods, enhanced multiplexing capabilities, and digital PCR for absolute quantification. In isothermal amplification, developments in LAMP, RPA, and CRISPR-based diagnostics have improved specificity and operational simplicity. Integration with microfluidics and portable devices has further expanded the scope of these technologies, particularly in resource-limited settings. Conclusion: The advancements in PCR and isothermal amplification technologies address critical challenges in diagnostics by improving accessibility, speed, and accuracy. Their continued development will likely transform molecular diagnostics, enabling precise and timely interventions across diverse applications.

Keywords: PCR, Isothermal Amplification, LAMP, RPA, CRISPR, Molecular Diagnostics, Point-of-Care Testing, Genomic Medicine.