ArticleThe Journal of molecular diagnostics : JMD2022
Characterizing the Impact of Primer-Template Mismatches on Recombinase Polymerase Amplification.
Article in The Journal of molecular diagnostics : JMD, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 24 citations in OpenAlex.
- RPA-CRISPR/Cas12a-Mediated Isothermal Amplification Technology for Visual Detection ofPlants (Basel, Switzerland) · 2026Article
- Simultaneous Detection of Human Norovirus GI, GII and Hepatitis A Virus Using CRISPR-Cas12a-Based RT-RPA and Lateral Flow Strip Method.Food and environmental virology · 2026Article
- CRISPR-based environmental detection of Burkholderia pseudomallei identifies sanitation gaps and melioidosis risk in northeast Thailand.Nature communications · 2026Article
- Iterative crRNA design and a PAM-free strategy enabled an ultra-specific RPA-CRISPR/Cas12a detection platform.Communications biology · 2024Article
- A Pan Plasmodium lateral flow recombinase polymerase amplification assay for monitoring malaria parasites in vectors and human populations.Scientific reports · 2024Article
- A multiplexed, allele-specific recombinase polymerase amplification assay with lateral flow readout for sickle cell disease detection.Lab on a chip · 2024Article
- Intrinsic RNA Targeting Triggers Indiscriminate DNase Activity of CRISPR-Cas12a.Angewandte Chemie (International ed. in English) · 2024Article
- Recombinase-based amplification coupled with lateral flow chromatography for the specific and sensitive detection and identification ofFrontiers in microbiology · 2024Article
- Improved Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the Rapid and Sensitive Detection ofViruses · 2023Article
Corrections and comments
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
Abstract
Recombinase polymerase amplification (RPA) is an isothermal amplification assay that has been ubiquitously utilized in the detection of infectious agents. Like any nucleic acid amplification technology, primer-template complementarity is critical to RPA reaction success. Mismatches arising in the primer-template complex are known to impact reaction kinetics, invalidate downstream analysis, such as nucleic acid quantification, and result in false negatives if used in a diagnostic capacity. Although the impact of specific primer-template mismatches has been well characterized for techniques such as PCR, characterization remains limited for RPA. Through our study, we systematically characterize the impact of mismatches on the RPA reaction, when located in the 3'-anchor region of the primer-template complex. Our investigation identified that the nucleotides involved, as well as position of each mismatch, influence the size of the impact, with terminal cytosine-thymine and guanine-adenine mismatches being the most detrimental. The presence of some mismatch combinations, such as a penultimate cytosine-cytosine and a terminal cytosine-adenine mismatch pairing, led to complete RPA reaction inhibition. Through the successful characterization of 315 mismatch combinations, researchers can optimize their RPA assay accordingly and seek to implement RPA technology for rapid, in-field genotyping.
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