A review of reaction enhancement strategies for isothermal nucleic acid amplification reactions

•Optimization requirements for isothermal nucleic acid amplification methods are highlighted.•The effects of nonspecific amplification on sensitivity and specificity are discussed, with an emphasis on decreasing nonspecific amplification in isothermal nucleic acid amplification methods.•Isothermal n...

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Bibliographic Details
Published inSensors and actuators reports Vol. 3; p. 100033
Main Authors Özay, Burcu, McCalla, Stephanie E
Format Journal Article
LanguageEnglish
Published Elsevier B.V 01.11.2021
Elsevier
Subjects
Diagnostics
Enzyme stability
Isothermal nucleic acid amplification reactions
Molecular detection
Nonspecific amplification
Sensitivity
Specificity
CRISPR
RPA
Molecular detection
Diagnostics
HDA
SSB-Single
RCA
Sensitivity
Specificity
Cas
Enzyme stability
NASBA
Nonspecific amplification
miRNA
LAMP
EXPAR
Isothermal nucleic acid amplification reactions
NSA
dsDNA
ssDNA
PCR
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Summary:•Optimization requirements for isothermal nucleic acid amplification methods are highlighted.•The effects of nonspecific amplification on sensitivity and specificity are discussed, with an emphasis on decreasing nonspecific amplification in isothermal nucleic acid amplification methods.•Isothermal nucleic acid amplification enhancement additives are summarized.•Potentially beneficial reaction additives that are not yet incorporated into isothermal amplification methods are discussed.•Potential adaptions of PCR enhancement additives to isothermal amplification are discussed. Isothermal nucleic acid amplification techniques are used to detect a variety of molecules such as RNAs, DNAs and proteins. When compared to the gold standard polymerase chain reaction, isothermal methods are rapid and simple but often have lower specificity and sensitivity. In this review, we provide a summary of reaction additives and techniques that enhance the sensitivity, specificity, stability, robustness, or speed of isothermal nucleic acid amplification reactions used for sensing applications. We also provide the mechanism and purpose of each enhancement technique, which can provide guidance when selecting reaction additives to optimize novel isothermal amplification systems and assays. [Display omitted]
ISSN:2666-0539
2666-0539
DOI:10.1016/j.snr.2021.100033