scholarly journals Impact of Dissociation Constant on the Detection Sensitivity of Polymerization-Based Signal Amplification Reactions

2013 ◽  
Vol 85 (17) ◽  
pp. 8055-8060 ◽  
Author(s):  
Kaja Kaastrup ◽  
Leslie Chan ◽  
Hadley D. Sikes
Keyword(s):  
Dissociation Constant ◽  
Signal Amplification ◽  
Detection Sensitivity

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.

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.

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.

Kinetically modulated specificity against single-base mutants in nucleic acid recycling circuitry using the destabilization motif

The Analyst ◽  
2017 ◽  
Vol 142 (15) ◽  
pp. 2786-2795
Author(s):  
Tsz Wing Fan ◽  
I-Ming Hsing
Keyword(s):  
Nucleic Acid ◽  
Signal Amplification ◽  
Detection Sensitivity ◽  
Single Base ◽  
Over Time

Signal amplification in nucleic acid sensing improves detection sensitivity, but difficulties remain in sustaining specificity over time, particularly under excess amounts of single-base mutants.

A signal amplifying fluorescent nanoprobe and lateral flow assay for ultrasensitive detection of cardiac biomarker troponin I

2019 ◽  
Vol 11 (28) ◽  
pp. 3506-3513 ◽  
Author(s):  
Doudou Lou ◽  
Lin Fan ◽  
Yongxin Ji ◽  
Ning Gu ◽  
Yu Zhang
Keyword(s):  
Signal Amplification ◽  
Troponin I ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Detection Sensitivity ◽  
Fluorescence Signal ◽  
Ultrasensitive Detection ◽  
Cardiac Biomarker ◽  
Fluorescent Nanoprobe ◽  
Fluorescence Signal Amplification

Novel functionalized nanoprobes based on a biotin–streptavidin system led to fluorescence signal amplification and the improvement of cTnI detection sensitivity.

New Advances in Super-Sensitive DNA-, RNA- and Antigen Detection: Combination of Labeled Tyramides with Nanogold-Silver Staining (NGSS)

1998 ◽  
Vol 4 (S2) ◽  
pp. 996-997
Author(s):  
G.W. Hacker ◽  
C. Hauser-Kronberger ◽  
I. Zehbe ◽  
H. Su ◽  
R. Tubbs
Keyword(s):  
Signal Amplification ◽  
Detection Efficiency ◽  
Detection Sensitivity ◽  
Silver Acetate ◽  
Rna Sequences ◽  
Silver Amplification ◽  
Specific Antigens ◽  
Gold Label ◽  
Catalyzed Reporter Deposition

In situ detection of specific antigens and nucleic acid sequences is possible with a variety of immunohistochemical (IHC) and in situ hybridization (ISH) methods. The drawback of conventional methodologies is their relatively low detection sensitivity. Improved detection efficiency is required in preparations containing only minute amounts of detectable antigens or only few copies of specific DNA and/or RNA sequences, which is often the case in routine paraffin cytoand histopathologic material. The use of gold as the label, followed by silver amplification (autometallography) avoids hazardous reagents and also gives an improved resolution compared to „standard” methods. Sensitivity problems may now be overcome by our specific combinations of recent technologies, utilitizing reporter molecule amplification with labeled tyramides (CARD, catalyzed reporter deposition, or tyramide signal amplification), followed by streptavidin bound to a new gold label, Nanogold™ (Nanoprobes, Stony Brook, NY), and silver acetate autometallography.

Applications of SIMS to electronic materials and devices

1992 ◽  
Vol 50 (2) ◽  
pp. 1682-1683
Author(s):  
S.F. Corcoran
Keyword(s):  
Depth Distribution ◽  
Secondary Ion Mass Spectrometry ◽  
Semiconductor Device ◽  
Electronic Materials ◽  
Profile Analysis ◽  
Semiconductor Industry ◽  
Small Diameter ◽  
Depth Resolution ◽  
Detection Sensitivity

Over the past decade secondary ion mass spectrometry (SIMS) has played an increasingly important role in the characterization of electronic materials and devices. The ability of SIMS to provide part per million detection sensitivity for most elements while maintaining excellent depth resolution has made this technique indispensable in the semiconductor industry. Today SIMS is used extensively in the characterization of dopant profiles, thin film analysis, and trace analysis in bulk materials. The SIMS technique also lends itself to 2-D and 3-D imaging via either the use of stigmatic ion optics or small diameter primary beams.By far the most common application of SIMS is the determination of the depth distribution of dopants (B, As, P) intentionally introduced into semiconductor materials via ion implantation or epitaxial growth. Such measurements are critical since the dopant concentration and depth distribution can seriously affect the performance of a semiconductor device. In a typical depth profile analysis, keV ion sputtering is used to remove successive layers the sample.

Plasminogen-Plasmin System IX. Specific Binding of Tranexamic Acid to Plasmin

1975 ◽  
Vol 33 (03) ◽  
pp. 573-585 ◽  
Author(s):  
Masahiro Iwamoto
Keyword(s):  
Tranexamic Acid ◽  
Affinity Chromatography ◽  
Dissociation Constant ◽  
Factor Xiii ◽  
Specific Binding ◽  
The Other ◽  
Inhibitory Mechanism ◽  
Binding Studies ◽  
Similar Affinity ◽  
Fibrin Structure

SummaryInteractions between tranexamic acid and protein were studied in respect of the antifibrinolytic actions of tranexamic acid. Tranexamic acid did neither show any interaction with fibrinogen or fibrin, nor was incorporated into cross-linked fibrin structure by the action of factor XIII. On the other hand, tranexamic acid bound to human plasmin with a dissociation constant of 3.5 × 10−5 M, which was very close to the inhibition constant (3.6 × 10−5 M) for this compound in inhibiting plasmin-induced fibrinolysis. The binding site of tranexamic acid on plasmin was not the catalytic site of plasmin, because TLCK-blocked plasmin also showed a similar affinity to tranexamic acid (the dissociation constant, 2.9–4.8 × 10−5 M).In the binding studies with the highly purified plasminogen and TLCK-plasmin preparations which were obtained by affinity chromatography on lysine-substituted Sepharose, the molar binding ratio was shown to be 1.5–1.6 moles tranexamic acid per one mole protein.On the basis of these and other findings, a model for the inhibitory mechanism of tranexamic acid is presented.

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