ArticleWorld journal of microbiology & biotechnology2019
Influence of design probe and sequence mismatches on the efficiency of fluorescent RPA.
Article in World journal of microbiology & biotechnology, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed.
- Establishment and clinical application of a rapid RPA-LFS assay for detectingFrontiers in cellular and infection microbiology · 2026Article
- Intrinsic RNA Targeting Triggers Indiscriminate DNase Activity of CRISPR-Cas12a.Angewandte Chemie (International ed. in English) · 2024Article
- Efficient detection of Streptococcus pyogenes based on recombinase polymerase amplification and lateral flow strip.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2024Article
- Rapid detection ofHeliyon · 2023Article
- Article
- Rapid and sensitive detection of pathogenicFish and shellfish immunology reports · 2022Article
- Rapid detection of human coronavirus NL63 by isothermal reverse transcription recombinase polymerase amplification.Journal of clinical virology plus · 2022Article
- Establishment and Clinical Application of a RPA-LFS Assay for Detection of Capsulated and Non-CapsulatedFrontiers in cellular and infection microbiology · 2022Article
- Rapid On-Site Detection of theFrontiers in plant science · 2022Article
- A Visualized Isothermal Amplification Method for Rapid and Specific Detection of Emetic and Non-emeticFrontiers in microbiology · 2022Article
- Rapid Detection ofFrontiers in cellular and infection microbiology · 2022Article
- Rapid detection of porcine circovirus type 4Frontiers in veterinary science · 2022Article
- Recombinase Polymerase Amplification Assay with and without Nuclease-Dependent-Labeled Oligonucleotide Probe.International journal of molecular sciences · 2021Article
- Minimally instrumented SHERLOCK (miSHERLOCK) for CRISPR-based point-of-care diagnosis of SARS-CoV-2 and emerging variants.Science advances · 2021Article
- Development and Application of Rapid Clinical Visualization Molecular Diagnostic Technology forFrontiers in cellular and infection microbiology · 2021Article
- Rapid Detection ofFrontiers in cellular and infection microbiology · 2021Article
- Development and evaluation of a rapid detection assay for severe fever with thrombocytopenia syndrome virus based on reverse-transcription recombinase polymerase amplification.Molecular and cellular probes · 2020Article
- A Recombinase Polymerase Amplification and Lateral Flow Strip Combined Method That DetectsFrontiers in microbiology · 2020Article
- Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection ofFoods (Basel, Switzerland) · 2019Article
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Authors and funding
11 authors.
Funding
Abstract
Recombinase polymerase amplification (RPA) is an isothermal amplification technique. Because of its short detection cycle and high specificity, it has been applied in various fields. However, the design of probe on the efficiency of RPA is not well understood and the effect of sequence mismatches of oligonucleotides on the performance of RPA is rarely discussed. In this study, we found that different primers with the same probe have a slight effect on the efficiency of fluorescent RPA, and different probes with the same amplified region have a great influence on the efficiency of fluorescent RPA. We summarized the design rules of probes suitable for fluorescent RPA by analyzing the experimental data. The rule is that the best distance between fluorescent groups in the probe is 1-2 bases, and the G content should be reduced as far as possible. In addition, we verified this rule by designing a series of probes. Furthermore, we found the base mismatches of the probe had a significant effect on RPA, which can lead to false positives and can change the amplification efficiency. However, 1-3 mismatches covering the center of the primer sequence only affect the amplification efficiency of RPA, not its specificity. And with an increase in the number of primer mismatches, the efficiency of RPA will decrease accordingly. This study suggests that the efficiency of fluorescent RPA is closely related to the probe. We recommend that when designing a fluorescent probe, one must consider the presence of closely related non-targets and specific bases.
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