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Most of these molecular methods are based on nucleic acid amplification (NAA).

Many molecular diagnostic methods have drastically evolved to sense nucleic acids in past decades (Martzy et al.

In addition, DNA and RNA show high affinity toward ions, signaling molecules like hormones and proteins, and even whole cells (Yan et al. This is possible as they show complementarity binding. Altogether, DNA and RNA are both important biomarkers for diagnostics and biological studies. The identification of viruses is critical to check substantial economic losses. Viruses are also essential molecules, as they play an important role in our life. This review comprehensively analyzed isothermal nucleic acid amplification methods, emphasizing their advantages, disadvantages and limitations.ĭNA, a unique molecule in each organism, signifies a promising recognition molecule for identifying living organisms. The coupling of isothermal amplification methods with advanced technologies such as CRISPR/Cas systems, fluorescence-based chemistries, microfluidics and paper-based sensors has significantly influenced the biosensing and diagnosis field. Using recombinant polymerases and other nucleic acid-modifying enzymes has dramatically broadened the detection range of target pathogens under the scanner. Furthermore, methods such as WGA, CPA, HDA, EXPAR, SMART, SPIA and DAMP were introduced for even more accuracy and robustness. LAMP, NASBA, SDA, RCA and RPA are some of the isothermal amplification techniques which are preferable for POC diagnostics. Isothermal amplification techniques have been found suitable for POC techniques and follow WHO’s ASSURED criteria. The isothermal amplification methods are exploited to develop miniaturized sensors against viruses, bacteria, fungi and other pathogenic organisms and have been applied for in situ diagnosis. However, using bulky thermal cyclers and costly laboratory setup limits its uses in developing countries, including India.

Efficient pathogen detection is a key to adequate food safety and hygiene. PCR, a gold standard technique for amplifying nucleic acids, is widely used in scientific research and diagnosis. Nucleic acids are prominent biomarkers for diagnosing infectious pathogens using nucleic acid amplification techniques (NAATs).