Emerging Alternatives to PCR for Efficient Nucleic Acid Amplification
Asim Debnath1*, Md Abu Bakar Siddique1, Nabil Deb Nath1, Md Abdur Rahman Biswash2, Tufael2
Journal of Primeasia 6(1) 1-8 https://doi.org/10.25163/primeasia.6110167
Submitted: 14 January 2025 Revised: 19 March 2025 Published: 22 March 2025
Abstract
Nucleic acid amplification is a vital part of molecular biology and biotechnology, and it serves a wide range of purposes, including clinical diagnostics, infectious disease detection, gene cloning, and quality control. While the polymerase chain reaction (PCR) has been the traditional go-to method for quite some time, it does come with some limitations. For instance, it requires thermal cycling, can be tricky with certain inhibitors, and relies on pricey equipment. To tackle these challenges, scientists have been excitedly working on alternative methods that can amplify nucleic acids without the need for complicated temperature changes. These isothermal techniques are not only easier and more cost-effective but also speed up the detection process. Some popular options include loop-mediated isothermal amplification (LAMP), nucleic acid sequence-based amplification (NASBA), strand displacement amplification (SDA), multiple displacement amplification (MDA), rolling circle amplification (RCA), and ligase chain reaction (LCR). Each of these methods brings something special to the table! For instance, LAMP is known for its awesome ability to provide visual results, while NASBA shines when it comes to detecting RNA. SDA and MDA are fantastic for amplifying small amounts of DNA, with MDA being particularly great for whole-genome amplification. RCA is super specific and resistant to contamination, and LCR is a go-to for spotting single-nucleotide variations. Even though these innovative methods have a lot of potential, many are still being fine-tuned for broader use in clinical and industrial settings. Luckily, advancements in real-time detection technologies, like molecular beacons and electrochemiluminescence (ECL), are making these methods even more reliable. As research continues to progress, these alternatives are set to complement or even take the place of PCR, offering faster, simpler, and more accessible solutions for a wide range of applications.
Keywords: Isothermal amplification, sensitivity, cost-effective, rapid detection, diagnostics, polymerase chain reaction alternatives
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