Inhibition of Non-specific Amplification in Loop-Mediated Isothermal Amplification via Tetramethylammonium Chloride

• Published in: Biochip journal Vol. 16; no. 3; pp. 326 - 333
• Main Authors: Jang, MinJu, Kim, Sanghyo
• Format: Journal Article
• Language: English
• Published: Seoul The Korean BioChip Society (KBCS) 01.09.2022; Springer Nature B.V; 한국바이오칩학회
• Subjects: Additives; Biomedical Engineering and Bioengineering; Biotechnology; Bovine serum albumin; Chemistry; Chemistry and Materials Science; Chlorides; Design; Dimethyl sulfoxide; DNA polymerase; Gene amplification; Original; Original Article; Point of care testing; Polymerase chain reaction; Primers; Protein structure; Redesign; Secondary structure; Serum albumin; Severe acute respiratory syndrome coronavirus 2; 생물공학; United States > US; South Korea; Seoul South Korea; Loop-mediated isothermal amplification; Additive; Non-specific amplification; Tetramethylammonium chloride
• ISSN: 1976-0280; 2092-7843
• DOI: 10.1007/s13206-022-00070-3

Loop-mediated isothermal amplification (LAMP) may be used in molecular and point-of-care diagnostics for pathogen detection. The amplification occurs under isothermal conditions using up to six primers. However, non-specific amplification is frequently observed in LAMP. Non-specific amplification has the potential to be triggered by forward and reverse internal primers. And the relatively low reaction temperature (55–65 °C) induces the secondary structure via primer–primer interactions. Primer redesign and probe design have been recommended to solve this problem. LAMP primers have strict conditions, such as Tm, GC contents, primer dimer, and distance between primers compared to conventional PCR primers. Probe design requires specialized knowledge to have high specificity for a target. In polymerase chain reaction (PCR), some chemicals or proteins are used for improving specificity and efficiency. Therefore, we hypothesized that additives can suppress the non-specific amplification. In this study, tetramethylammonium chloride (TMAC), formamide, dimethyl sulfoxide, Tween 20, and bovine serum albumin have been used as LAMP additives. In our study, TMAC was presented as a promising additive for suppressing non-specific amplification in LAMP.