scholarly journals Centrifugal step emulsification applied for absolute quantification of nucleic acids by digital droplet RPA

Lab on a Chip ◽  
2015 ◽  
Vol 15 (13) ◽  
pp. 2759-2766 ◽  
Author(s):  
Friedrich Schuler ◽  
Frank Schwemmer ◽  
Martin Trotter ◽  
Simon Wadle ◽  
Roland Zengerle ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Absolute Quantification ◽  
Recombinase Polymerase Amplification ◽  
Dead Volume

Centrifugal step emulsification enables dead volume free emulsification, it was applied to the first digital droplet recombinase polymerase amplification (RPA).

scholarly journals Picoliter Well Array Chip-Based Digital Recombinase Polymerase Amplification for Absolute Quantification of Nucleic Acids

PLoS ONE ◽  
2016 ◽  
Vol 11 (4) ◽  
pp. e0153359 ◽  
Author(s):  
Zhao Li ◽  
Yong Liu ◽  
Qingquan Wei ◽  
Yuanjie Liu ◽  
Wenwen Liu ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Absolute Quantification ◽  
Recombinase Polymerase Amplification

scholarly journals Comparative Use of Quantitative PCR (qPCR), Droplet Digital PCR (ddPCR), and Recombinase Polymerase Amplification (RPA) in the Detection of Shiga Toxin-Producing E. coli (STEC) in Environmental Samples

Water ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 3507
Author(s):  
Mark A. Ibekwe ◽  
Shelton E. Murinda ◽  
Stanley Park ◽  
Amarachukwu Obayiuwana ◽  
Marcia A. Murry ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
Environmental Samples ◽  
Point Of Care ◽  
Foodborne Pathogen ◽  
Digital Pcr ◽  
Absolute Quantification ◽  
Droplet Digital Pcr ◽  
Recombinase Polymerase Amplification ◽  
High Rate ◽  
E Coli

E. coli O157:H7 is a foodborne pathogen that constitutes a global threat to human health. However, the quantification of this pathogen in food and environmental samples may be problematic at the low cell numbers commonly encountered in environmental samples. In this study, we used recombinase polymerase amplification (RPA) for the detection of E. coli O157:H7, real-time quantitative PCR (qPCR) for quantification, and droplet digital PCR (ddPCR) for absolute and accurate quantification of E. coli O157:H7 from spiked and environmental samples. Primer and probe sets were used for the detection of stx1 and stx2 using RPA. Genes encoding for stx1, stx2, eae, and rfbE were used to quantify E. coli O157:H7 in the water samples. Furthermore, duplex ddPCR assays were used to quantify the pathogens in these samples. Duplex assay set 1 used stx1 and rfbE genes, while assay set 2 used stx2 and eae genes. Droplet digital PCR was used for the absolute quantification of E. coli O15:H7 in comparison with qPCR for the spiked and environmental samples. The RPA results were compared to those from qPCR and ddPCR in order to assess the efficiency of the RPA compared with the PCR methods. The assays were further applied to the dairy lagoon effluent (DLE) and the high rate algae pond (HRAP) effluent, which were fed with diluted DLE. The RPA detected was <10 CFU/mL, while ddPCR showed quantification from 1 to 104 CFU/mL with a high reproducibility. In addition, quantification by qPCR was from 103 to 107 CFU/mL of the wastewater samples. Therefore, the RPA assay has potential as a point of care tool for the detection of E. coli O157:H7 from different environmental sources, followed by quantification of the target concentrations.

Ultra-fast detection and quantification of nucleic acids by amplification-free fluorescence assay

The Analyst ◽  
2020 ◽  
Vol 145 (17) ◽  
pp. 5836-5844
Author(s):  
Jesper Uhd ◽  
Laura Miotke ◽  
Hanlee P. Ji ◽  
Marina Dunaeva ◽  
Ger J. M. Pruijn ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Absolute Quantification ◽  
Fluorescence Assay ◽  
Fast Detection ◽  
Dna And Rna ◽  
Detection And Quantification

Fast and reliable assay for amplification-free absolute quantification of DNA and RNA in biofluids.

Generating Microdroplet Array on Photonic Pseudo-paper for Absolute Quantification of Nucleic Acids

2018 ◽  
Vol 10 (45) ◽  
pp. 39144-39150 ◽  
Author(s):  
Junjie Chi ◽  
Changmin Shao ◽  
Xin Du ◽  
Hong Liu ◽  
Zhongze Gu
Keyword(s):  
Nucleic Acids ◽  
Absolute Quantification
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