Enzyme-Assisted Nucleic Acid Amplification in Molecular Diagnosis: A Review

Abstract

Infectious diseases and tumors have become the biggest medical challenges in the 21st century. They are driven by multiple factors such as population growth, aging, climate change, genetic predispositions and more. Nucleic acid amplification technologies (NAATs) are used for rapid and accurate diagnostic testing, providing critical information in order to facilitate better follow-up treatment and prognosis. NAATs are widely used due their high sensitivity, specificity, rapid amplification and detection. It should be noted that different NAATs can be selected according to different environments and research fields; for example, isothermal amplification with a simple operation can be preferred in developing countries or resource-poor areas. In the field of translational medicine, CRISPR has shown great prospects. The core component of NAAT lies in the activity of different enzymes. As the most critical material of nucleic acid amplification, the key role of the enzyme is self-evident, playing the upmost important role in molecular diagnosis. In this review, several common enzymes used in NAATs are compared and described in detail. Furthermore, we summarize both the advances and common issues of NAATs in clinical application.

Advancements in magnetic nanoparticle-based biosensors for point-of-care testing

“…Factors that can affect the detection of analytes in a sample include concentration, separation, enrichment, and derivatization. In essence, sample preparation refers to the process of eliminating interfering substances from biological samples, thereby reducing their inherent complexity ( Wang et al, 2023 ). The primary goal is to enhance the accuracy, sensitivity, and selectivity of subsequent analytical measurements and analyses by selectively removing or minimizing interferences, such as matrix components, contaminants, or co-existing analytes.…”

The Role of Gut Microbiota in the Clinical Outcome of Septic Patients: State of the Art and Future Perspectives

“…The detection of bacterial DNA fragments via real-time polymerase chain reaction (RT-PCR) in blood samples and the detection of 16S rRNA fragments of Gram-positive and Gram-negative bacteria or 18S rRNA fragments of Candida spp. seem to be very promising for shortening pathogen identification, as they have shown a high degree of specificity and sensitivity, thus reducing mortality, length of hospitalization, and ICU stay of patients [20]. However, these techniques are not sufficient to differentiate sepsis from other inflammatory processes, and there are no biomarkers able to identify only septic patients [11].…”

Multiplexed electrical detection of whole viruses from plasma in a microfluidic platform

“…Much research is being done on other isothermal nucleic acid amplification techniques such as loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) as alternatives to PCR to help alleviate this issue, however, these techniques have limited ability to acquire true simultaneous multiplexing without partitioning samples into multiple individual reactions that are performed separately, or utilizing fluorophores with different colors that may be hard to distinguish on a PoC device. [2][3][4][5] The other option is immunoassays, which can be used to detect either anti-viral antibodies or viral antigens. 1 The current gold standard for immunoassays are the enzyme-linked immunosorbent assays (ELISA), however these can take several hours to complete and requires bulky laboratory equipment making it hard to adapt for PoC applications and limited in its ability to achieve a speedy diagnosis.…”