Simple and sensitive microbial pathogen detection using a label-free DNA amplification assay

2016 ◽  
Vol 52 (47) ◽  
pp. 7505-7508 ◽  
Author(s):  
Yuhuan Sun ◽  
Chuanqi Zhao ◽  
Zhengqing Yan ◽  
Jinsong Ren ◽  
Xiaogang Qu
Keyword(s):  
Magnetic Nanoparticles ◽  
Pathogen Detection ◽  
Dna Amplification ◽  
Label Free ◽  
Exonuclease Iii ◽  
Microbial Pathogen ◽  
Free Dna ◽  
Amplification Assay

A simple and facile strategy for sensitive pathogen detection has been developed by a combination of quaternized magnetic nanoparticles and a label-free exonuclease III-assisted DNA amplification assay.

Label-free DNA detection based on oligonucleotide-stabilized silver nanoclusters and exonuclease III-catalyzed target recycling amplification

2014 ◽  
Vol 6 (15) ◽  
pp. 6082-6087 ◽  
Author(s):  
Hui Ma ◽  
Wei Wei ◽  
Qian Lu ◽  
Zhixin Zhou ◽  
Henan Li ◽  
...  
Keyword(s):  
High Sensitivity ◽  
Silver Nanoclusters ◽  
Label Free ◽  
Exonuclease Iii ◽  
Target Recycling ◽  
Free Dna ◽  
Sensitivity And Selectivity ◽  
Exo Iii ◽  
Ag Ncs ◽  
Recycling Amplification

A label-free DNA biosensor with high sensitivity and selectivity is constructed by using DNA–Ag NCs and Exo III-catalyzed target recycling amplification.

A cascade signal amplification strategy for sensitive and label-free DNA detection based on Exo III-catalyzed recycling coupled with rolling circle amplification

The Analyst ◽  
2014 ◽  
Vol 139 (11) ◽  
pp. 2884-2889 ◽  
Author(s):  
Xingti Liu ◽  
Qingwang Xue ◽  
Yongshun Ding ◽  
Jing Zhu ◽  
Lei Wang ◽  
...  
Keyword(s):  
Detection Method ◽  
Dna Detection ◽  
Rolling Circle Amplification ◽  
Label Free ◽  
Exonuclease Iii ◽  
Rolling Circle ◽  
Free Dna ◽  
Exo Iii ◽  
Amplification Strategy ◽  
Cascade Amplification

A sensitive and label-free DNA detection method was developed based on cascade amplification combining exonuclease-III recycling with rolling circle amplification.

A label-free signal amplification assay for DNA detection based on exonuclease III and nucleic acid dye SYBR Green I

Talanta ◽  
2013 ◽  
Vol 114 ◽  
pp. 49-53 ◽  
Author(s):  
Aihua Zheng ◽  
Ming Luo ◽  
Dongshan Xiang ◽  
Xia Xiang ◽  
Xinghu Ji ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Signal Amplification ◽  
Dna Detection ◽  
Sybr Green ◽  
Sybr Green I ◽  
Label Free ◽  
Acid Dye ◽  
Exonuclease Iii ◽  
Amplification Assay

scholarly journals Development of a Sensitive and Rapid Recombinase Polymerase Amplification Assay for Detection of Anaplasma phagocytophilum

2020 ◽  
Vol 58 (5) ◽  
Author(s):  
Le Jiang ◽  
Philip Ching ◽  
Chien-Chung Chao ◽  
J. Stephen Dumler ◽  
Wei-Mei Ching
Keyword(s):  
Anaplasma Phagocytophilum ◽  
Pathogenic Bacteria ◽  
Pathogen Detection ◽  
Limit Of Detection ◽  
Dna Amplification ◽  
Recombinase Polymerase Amplification ◽  
Detection Methods ◽  
Content Type ◽  
Amplification Assay ◽  
Nonspecific Amplification

ABSTRACT Human granulocytic anaplasmosis (HGA) is a tick-borne disease caused by the obligate intracellular Gram-negative bacterium Anaplasma phagocytophilum. The disease often presents with nonspecific symptoms with negative serology during the acute phase. Direct pathogen detection is the best approach for early confirmatory diagnosis. Over the years, PCR-based molecular detection methods have been developed, but optimal sensitivity is not achieved by conventional PCR while real-time PCR requires expensive and sophisticated instruments. To improve the sensitivity and also develop an assay that can be used in resource-limited areas, an isothermal DNA amplification assay based on recombinase polymerase amplification (RPA) was developed. To do this, we identified a 171-bp DNA sequence within multiple paralogous copies of msp2 within the genome of A. phagocytophilum. Our novel RPA assay targeting this sequence has an analytical limit of detection of one genome equivalent copy of A. phagocytophilum and can reliably detect 125 bacteria/ml in human blood. A high level of specificity was demonstrated by the absence of nonspecific amplification using genomic DNA from human or DNA from other closely-related pathogenic bacteria, such as Anaplasma platys, Ehrlichia chaffeensis, Orientia tsutsugamushi, and Rickettsia rickettsii, etc. When applied to patient DNA extracted from whole blood, this new RPA assay was able to detect 100% of previously diagnosed A. phagocytophilum cases. The sensitivity and rapidness of this assay represents a major improvement for early diagnosis of A. phagocytophilum in human patients and suggest a role for better surveillance in its reservoirs or vectors, especially in remote regions where resources are limited.

Biochip for DNA Amplification and Label-free DNA Detection

2007 ◽  
Author(s):  
G. Hairer ◽  
M.J. Vellekoop ◽  
M.H. Mansfeld ◽  
C. Nohammer
Keyword(s):  
Dna Detection ◽  
Dna Amplification ◽  
Label Free ◽  
Free Dna

Label-free DNA Y junction for detection of Hg2+ using exonuclease III or graphene oxide-assisted background reduction

2019 ◽  
Vol 145 ◽  
pp. 1086-1093 ◽  
Author(s):  
Lili Yu ◽  
Hui Xu ◽  
Hou Chen ◽  
Shengxiao Zhang ◽  
Tingting Jiang ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Label Free ◽  
Exonuclease Iii ◽  
Background Reduction ◽  
Free Dna

scholarly journals Label-Free Electrochemical Sensor for Rapid Bacterial Pathogen Detection Using Vancomycin-Modified Highly Branched Polymers

Sensors ◽  
2021 ◽  
Vol 21 (5) ◽  
pp. 1872
Author(s):  
Holger Schulze ◽  
Harry Wilson ◽  
Ines Cara ◽  
Steven Carter ◽  
Edward N. Dyson ◽  
...  
Keyword(s):  
Pathogen Detection ◽  
Electrochemical Impedance ◽  
Point Of Care ◽  
Branched Polymers ◽  
Label Free ◽  
Conformation Change ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Staphylococcus Carnosus ◽  
Label Free Detection

Rapid point of care tests for bacterial infection diagnosis are of great importance to reduce the misuse of antibiotics and burden of antimicrobial resistance. Here, we have successfully combined a new class of non-biological binder molecules with electrochemical impedance spectroscopy (EIS)-based sensor detection for direct, label-free detection of Gram-positive bacteria making use of the specific coil-to-globule conformation change of the vancomycin-modified highly branched polymers immobilized on the surface of gold screen-printed electrodes upon binding to Gram-positive bacteria. Staphylococcus carnosus was detected after just 20 min incubation of the sample solution with the polymer-functionalized electrodes. The polymer conformation change was quantified with two simple 1 min EIS tests before and after incubation with the sample. Tests revealed a concentration dependent signal change within an OD600 range of Staphylococcus carnosus from 0.002 to 0.1 and a clear discrimination between Gram-positive Staphylococcus carnosus and Gram-negative Escherichia coli bacteria. This exhibits a clear advancement in terms of simplified test complexity compared to existing bacteria detection tests. In addition, the polymer-functionalized electrodes showed good storage and operational stability.

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