scholarly journals 2′-O-Methyl molecular beacon: a promising molecular tool that permits elimination of sticky-end pairing and improvement of detection sensitivity

RSC Advances ◽  
2020 ◽  
Vol 10 (68) ◽  
pp. 41618-41624
Author(s):  
Jiafeng Gao ◽  
Yang Li ◽  
Wenqin Li ◽  
Chaofei Zeng ◽  
Fengna Xi ◽  
...  
Keyword(s):  
Signal Amplification ◽  
Molecular Beacon ◽  
Dnase I ◽  
Fluorescence Sensor ◽  
Detection Sensitivity ◽  
Pairing Effect ◽  
Molecular Tool ◽  
Sticky End

A novel fluorescence sensor is reported based on the employment of an intriguing 2′-O-methyl molecular beacon (MB) and DNase I, the coupled use of which is responsible for both the elimination of the sticky-end pairing effect and signal amplification capability.

scholarly journals SARS-CoV-2 RNA Detection with Duplex-Specific Nuclease Signal Amplification

Micromachines ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 197
Author(s):  
Meiqing Liu ◽  
Haoran Li ◽  
Yanwei Jia ◽  
Pui-In Mak ◽  
Rui P. Martins
Keyword(s):  
Signal Amplification ◽  
Incubation Temperature ◽  
Dna Probes ◽  
Detection Sensitivity ◽  
N Gene ◽  
Rna Detection ◽  
Gold Standard Method ◽  
Complementary Method ◽  
Amplification Method ◽  
Virus Rna

The emergence of the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a zoonotic pathogen, has led to the outbreak of coronavirus disease 2019 (COVID-19) pandemic and brought serious threats to public health worldwide. The gold standard method for SARS-CoV-2 detection requires both reverse transcription (RT) of the virus RNA to cDNA and then polymerase chain reaction (PCR) for the cDNA amplification, which involves multiple enzymes, multiple reactions and a complicated assay optimization process. Here, we developed a duplex-specific nuclease (DSN)-based signal amplification method for SARS-CoV-2 detection directly from the virus RNA utilizing two specific DNA probes. These specific DNA probes can hybridize to the target RNA at different locations in the nucleocapsid protein gene (N gene) of SARS-CoV-2 to form a DNA/RNA heteroduplex. DSN cleaves the DNA probe to release fluorescence, while leaving the RNA strand intact to be bound to another available probe molecule for further cleavage and fluorescent signal amplification. The optimized DSN amount, incubation temperature and incubation time were investigated in this work. Proof-of-principle SARS-CoV-2 detection was demonstrated with a detection sensitivity of 500 pM virus RNA. This simple, rapid, and direct RNA detection method is expected to provide a complementary method for the detection of viruses mutated at the PCR primer-binding regions for a more precise detection.

scholarly journals Highly sensitive and multiplexed in situ protein profiling with cleavable fluorescent streptavidin

2019 ◽  
Author(s):  
Renjie Liao ◽  
Diego Mastroeni ◽  
Paul D. Coleman ◽  
Jia Guo
Keyword(s):  
Signal Amplification ◽  
Individual Cell ◽  
Protein Detection ◽  
Protein Profiling ◽  
Detection Sensitivity ◽  
Layer By Layer ◽  
Protein Targets ◽  
Highly Sensitive ◽  
Wide Range

AbstractThe ability to perform highly sensitive and multiplexed in situ protein analysis is crucial to advance our understanding of normal physiology and disease pathogenesis. To achieve this goal, here we develop an approach using cleavable biotin conjugated antibodies and cleavable fluorescent streptavidin (CFS). In this approach, protein targets are first recognized by the cleavable biotin labeled antibodies. Subsequently, CFS is applied to stain the protein targets. Though layer-by-layer signal amplification using cleavable biotin conjugated orthogonal antibodies and CSF, the protein detection sensitivity can be enhanced by at least 10 fold, compared with the existing methods. After imaging, the fluorophores and the biotins unbound to streptavidin are removed by chemical cleavage. The leftover streptavidin is blocked by biotin. Upon reiterative analysis cycles, a large number of different proteins with a wide range of expression levels can be unambiguously detected in individual cell in situ.

Enhancing the sensitivity of colorimetric lateral flow assay (CLFA) through signal amplification techniques

2018 ◽  
Vol 6 (44) ◽  
pp. 7102-7111 ◽  
Author(s):  
Haihang Ye ◽  
Xiaohu Xia
Keyword(s):  
Signal Amplification ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Detection Sensitivity

This article highlights recent signal amplification techniques for enhancing the detection sensitivity of colorimetric lateral flow assay.

Highly sensitive and selective detection of miRNA: DNase I-assisted target recycling using DNA probes protected by polydopamine nanospheres

2015 ◽  
Vol 51 (11) ◽  
pp. 2156-2158 ◽  
Author(s):  
Yi Xie ◽  
Xiaoyan Lin ◽  
Yishun Huang ◽  
Rujun Pan ◽  
Zhi Zhu ◽  
...  
Keyword(s):  
Signal Amplification ◽  
Dnase I ◽  
Dna Probes ◽  
Selective Detection ◽  
Protective Properties ◽  
Target Recycling ◽  
Amplification Method ◽  
Highly Sensitive ◽  
Nuclease Digestion

Based on the protective properties of polydopamine nanospheres for DNA probes against nuclease digestion, we have developed a DNase I-assisted target recycling signal amplification method for highly sensitive and selective detection of miRNA.

A highly sensitive homogeneous electrochemical assay for alkaline phosphatase activity based on single molecular beacon-initiated T7 exonuclease-mediated signal amplification

The Analyst ◽  
2015 ◽  
Vol 140 (12) ◽  
pp. 4030-4036 ◽  
Author(s):  
Lianfang Zhang ◽  
Ting Hou ◽  
Haiyin Li ◽  
Feng Li
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Signal Amplification ◽  
Molecular Beacon ◽  
Electrochemical Assay ◽  
Highly Sensitive ◽  
T7 Exonuclease

A highly sensitive homogeneous electrochemical assay for alkaline phosphatase activity based on single molecular beacon-initiated T7 exonuclease-assisted signal amplification.

scholarly journals Lateral Flow Immunoassay Coupled with Copper Enhancement for Rapid and Sensitive SARS-CoV-2 Nucleocapsid Protein Detection

Biosensors ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 13
Author(s):  
Tao Peng ◽  
Xueshima Jiao ◽  
Zhanwei Liang ◽  
Hongwei Zhao ◽  
Yang Zhao ◽  
...  
Keyword(s):  
Signal Amplification ◽  
Nucleocapsid Protein ◽  
High Efficiency ◽  
Protein Detection ◽  
Antigen Detection ◽  
Lateral Flow ◽  
Copper Deposition ◽  
Detection Sensitivity ◽  
Rapid Screening ◽  
Lateral Flow Immunoassay

The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory coronavirus 2 (SARS-CoV-2) is still raging all over the world. Hence, the rapid and sensitive screening of the suspected population is in high demand. The nucleocapsid protein (NP) of SARS-CoV-2 has been selected as an ideal marker for viral antigen detection. This study describes a lateral flow immunoassay (LFIA) based on colloidal gold nanoparticles for rapid NP antigen detection, in which sensitivity was improved through copper deposition-induced signal amplification. The detection sensitivity of the developed LFIA for NP antigen detection (using certified reference materials) under the optimized parameters was 0.01 μg/mL and was promoted by three orders of magnitude to 10 pg/mL after copper deposition signal amplification. The LFIA coupled with the copper enhancement technique has many merits such as low cost, high efficiency, and high sensitivity. It provides an effective approach to the rapid screening, diagnosis, and monitoring of the suspected population in the COVID-19 outbreak.

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