Signal-on electrochemical DNA (E-DNA) sensor for accurate quantification of nicking-assisted rolling circle amplification (N-RCA) products with attomolar sensitivity

2021 ◽  
Author(s):  
Mengmeng Li ◽  
Dandan Li ◽  
Guidan Huang ◽  
Linying Zhou ◽  
Qilin Wen ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Molecular Diagnosis ◽  
Rolling Circle Amplification ◽  
High Specificity ◽  
Dna Sensor ◽  
Rolling Circle ◽  
Specificity And Sensitivity ◽  
Nucleic Acid Analysis ◽  
Amplification Technique ◽  
Accurate Quantification

Rolling circle amplification (RCA) has become increasingly important amplification technique in nucleic acid analysis, immunoassay, and molecular diagnosis due to its high specificity and sensitivity. However, the accurate quantification of...

A netlike rolling circle nucleic acid amplification technique

The Analyst ◽  
2015 ◽  
Vol 140 (1) ◽  
pp. 74-78 ◽  
Author(s):  
Xiaoli Zhu ◽  
Chang Feng ◽  
Bin Zhang ◽  
Hui Tong ◽  
Tao Gao ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Rolling Circle Amplification ◽  
Nucleic Acid Amplification ◽  
Amplification Efficiency ◽  
Rolling Circle ◽  
Isothermal Nucleic Acid Amplification ◽  
Amplification Technique ◽  
Nucleic Acid Amplification Technique ◽  
Network Morphology

An isothermal nucleic acid amplification technique termed as netlike rolling circle amplification is proposed. Dense and uniform network morphology of amplified products is first observed, suggesting the ultrahigh amplification efficiency.

scholarly journals Multiplex real-time PCR assay combined with rolling circle amplification (MPRP) using universal primers for non-invasive detection of tumor-related mutations

RSC Advances ◽  
2018 ◽  
Vol 8 (48) ◽  
pp. 27375-27381 ◽  
Author(s):  
Jian Gong ◽  
Yishuai Li ◽  
Ting Lin ◽  
Xiaoyan Feng ◽  
Li Chu
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rolling Circle Amplification ◽  
High Specificity ◽  
Universal Primers ◽  
Multiplex Detection ◽  
Pcr Assay ◽  
Rolling Circle ◽  
Non Invasive ◽  
Specificity And Sensitivity

The MPRP system for SNP discrimination was developed, which showed high specificity and sensitivity for multiplex detection of tumor-related mutations.

The analysis of proteins and small molecules based on sterically tunable nucleic acid hyperbranched rolling circle amplification

2017 ◽  
Vol 91 ◽  
pp. 136-142 ◽  
Author(s):  
Hai Shi ◽  
Xiaoxia Mao ◽  
Xiaoxia Chen ◽  
Zihan Wang ◽  
Keming Wang ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Small Molecules ◽  
Rolling Circle Amplification ◽  
Rolling Circle

Mitigating the Non-Specific Amplification in RCA: Assessment of the Role of Ligation and Exonuclease Digestion in Circular DNA Preparation

2021 ◽  
Author(s):  
Vandana Kuttappan Nair ◽  
Chandrika Sharma ◽  
Mrittika Sengupta ◽  
Souradyuti Ghosh
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
False Positive ◽  
Rolling Circle Amplification ◽  
Circular Dna ◽  
Rolling Circle ◽  
Specific Amplification ◽  
Exonuclease Digestion ◽  
Sticky End

<b>Layman Summary: </b>Rolling circle amplification (RCA) is a popular and extensively used bioanalytical tool. Like any nucleic acid amplifications, non-specific amplification may occur in it and risk generating false positive readouts. The work described in the manuscript investigates non-specific amplification in RCA as a function of ligation and exonuclease digestion assays during the synthesis of circular DNA. In particular, it investigates and compares the role of three different ligation techniques, namely splint-padlock ligation, cohesive end (sticky end ligation), and self-annealing ligation. In addition, it also probes the role of single exonuclease vs dual exonuclease digestions. We employed real time fluorescence to quantify the effect of these factors. Finally, our work hypothesizes the possible origins of non-specific amplification in RCA.

scholarly journals Padlock Probe Assay for Detection and Subtyping of Seasonal Influenza

2018 ◽  
Vol 64 (12) ◽  
pp. 1704-1712 ◽  
Author(s):  
Felix Neumann ◽  
Iván Hernández-Neuta ◽  
Malin Grabbe ◽  
Narayanan Madaboosi ◽  
Jan Albert ◽  
...  
Keyword(s):  
Seasonal Influenza ◽  
Rolling Circle Amplification ◽  
High Specificity ◽  
Clinical Samples ◽  
Padlock Probe ◽  
Rolling Circle ◽  
Diagnostic Assays ◽  
Padlock Probes ◽  
Probe Assay ◽  
Digital Quantification

Abstract BACKGROUND Influenza remains a constant threat worldwide, and WHO estimates that it affects 5% to 15% of the global population each season, with an associated 3 to 5 million severe cases and up to 500000 deaths. To limit the morbidity and the economic burden of influenza, improved diagnostic assays are needed. METHODS We developed a multiplexed assay for the detection and subtyping of seasonal influenza based on padlock probes and rolling circle amplification. The assay simultaneously targets all 8 genome segments of the 4 circulating influenza variants—A(H1N1), A(H3N2), B/Yamagata, and B/Victoria—and was combined with a prototype cartridge for inexpensive digital quantification. Characterized virus isolates and patient nasopharyngeal swabs were used for assay design and analytical validation. The diagnostic performance was assessed by blinded testing of 50 clinical samples analyzed in parallel with a commercial influenza assay, Simplexa™ Flu A/B & RSV Direct. RESULTS The assay had a detection limit of 18 viral RNA copies and achieved 100% analytical and clinical specificity for differential detection and subtyping of seasonal circulating influenza variants. The diagnostic sensitivity on the 50 clinical samples was 77.5% for detecting influenza and up to 73% for subtyping seasonal variants. CONCLUSIONS We have presented a proof-of-concept padlock probe assay combined with an inexpensive digital readout for the detection and subtyping of seasonal influenza strains A and B. The demonstrated high specificity and multiplexing capability, together with the digital quantification, established the assay as a promising diagnostic tool for seasonal influenza.

Nucleic acid sensing by regenerable surface-associated isothermal rolling circle amplification

2007 ◽  
Vol 22 (7) ◽  
pp. 1236-1244 ◽  
Author(s):  
Erik L. McCarthy ◽  
Lee E. Bickerstaff ◽  
Mauricio Pereira da Cunha ◽  
Paul J. Millard
Keyword(s):  
Nucleic Acid ◽  
Rolling Circle Amplification ◽  
Rolling Circle
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