DNA-templated copper nanoclusters obtained via TdT isothermal nucleic acid amplification for mercury(ii) assay

2019 ◽  
Vol 11 (32) ◽  
pp. 4165-4172 ◽  
Author(s):  
Jing-Lin He ◽  
Xing-Xing Wang ◽  
Ting-Ting Mei ◽  
Ling Wu ◽  
Ju-Lan Zeng ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Fluorescence Sensor ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Copper Nanoclusters ◽  
Isothermal Nucleic Acid Amplification

The Hg2+ fluorescence sensor based on TdT isothermal amplification DNA templated CuNCs was successfully constructed.

Long-term dry storage of enzyme-based reagents for isothermal nucleic acid amplification in a porous matrix for use in point-of-care diagnostic devices

The Analyst ◽  
2020 ◽  
Vol 145 (21) ◽  
pp. 6875-6886 ◽  
Author(s):  
Sujatha Kumar ◽  
Ryan Gallagher ◽  
Josh Bishop ◽  
Enos Kline ◽  
Joshua Buser ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Glass Fiber ◽  
Point Of Care ◽  
Porous Matrix ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Isothermal Nucleic Acid Amplification ◽  
Dry Storage

Long-term dry storage of enzyme-based isothermal amplification reagents in glass fiber porous matrix for use in point-of-care devices.

A novel exonuclease-assisted isothermal nucleic acid amplification with ultrahigh specificity mediated by full-length Bst DNA polymerase

2018 ◽  
Vol 54 (75) ◽  
pp. 10562-10565 ◽  
Author(s):  
Xin Ye ◽  
Yang Li ◽  
Lijuan Wang ◽  
Xueen Fang ◽  
Jilie Kong
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Dna Polymerase ◽  
Isothermal Amplification ◽  
Full Length ◽  
Nucleic Acid Amplification ◽  
Isothermal Nucleic Acid Amplification

A novel exonuclease-assisted isothermal amplification to amplify and determine nucleic acids very sensitively and with ultrahigh specificity.

scholarly journals High-Surety Isothermal Amplification and Detection of SARS-CoV-2

mSphere ◽  
2021 ◽  
Vol 6 (3) ◽  
Author(s):  
Sanchita Bhadra ◽  
Timothy E. Riedel ◽  
Simren Lakhotia ◽  
Nicholas D. Tran ◽  
Andrew D. Ellington
Keyword(s):  
Nucleic Acid ◽  
False Negative ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Boolean Logic ◽  
Nucleic Acid Amplification ◽  
Rapid Diagnostics ◽  
One Pot ◽  
Loop Mediated Isothermal Amplification ◽  
Isothermal Nucleic Acid Amplification

ABSTRACT Isothermal nucleic acid amplification tests (iNATs), such as loop-mediated isothermal amplification (LAMP), are good alternatives to PCR-based amplification assays, especially for point-of-care and low-resource use, in part because they can be carried out with relatively simple instrumentation. However, iNATs can often generate spurious amplicons, especially in the absence of target sequences, resulting in false-positive results. This is especially true if signals are based on non-sequence-specific probes, such as intercalating dyes or pH changes. In addition, pathogens often prove to be moving, evolving targets and can accumulate mutations that will lead to inefficient primer binding and thus false-negative results. Multiplex assays targeting different regions of the analyte and logical signal readout using sequence-specific probes can help to reduce both false negatives and false positives. Here, we describe rapid conversion of three previously described SARS-CoV-2 LAMP assays that relied on a non-sequence-specific readout into individual and multiplex one-pot assays that can be visually read using sequence-specific oligonucleotide strand exchange (OSD) probes. We describe both fluorescence-based and Boolean logic-gated colorimetric lateral flow readout methods and demonstrate detection of SARS-CoV-2 virions in crude human saliva. IMPORTANCE One of the key approaches to treatment and control of infectious diseases, such as COVID-19, is accurate and rapid diagnostics that is widely deployable in a timely and scalable manner. To achieve this, it is essential to go beyond the traditional gold standard of quantitative PCR (qPCR) that is often faced with difficulties in scaling due to the complexity of infrastructure and human resource requirements. Isothermal nucleic acid amplification methods, such as loop-mediated isothermal amplification (LAMP), have been long pursued as ideal, low-tech alternatives for rapid, portable testing. However, isothermal approaches often suffer from false signals due to employment of nonspecific readout methods. We describe general principles for rapidly converting nonspecifically read LAMP assays into assays that are read in a sequence-specific manner by using oligonucleotide strand displacement (OSD) probes. We also demonstrate that inclusion of OSD probes in LAMP assays maintains the simplicity of one-pot assays and a visual yes/no readout by using fluorescence or colorimetric lateral-flow dipsticks while providing accurate sequence-specific readout and the ability to logically query multiplex amplicons for redundancy or copresence. These principles not only yielded high-surety isothermal assays for SARS-CoV-2 but might also aid in the design of more sophisticated molecular assays for other analytes.

scholarly journals Strategies for Isothermal Amplification of Nucleic Acids: are they Ready to be Applied in Point of Care Diagnosis of Mycosis?

2020 ◽  
Vol 11 (3) ◽  
pp. 10559-10571
Keyword(s):  
Nucleic Acid ◽  
Clinical Laboratory ◽  
Multiple Displacement Amplification ◽  
Rolling Circle Amplification ◽  
High Sensitivity ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Clinical Samples ◽  
Rolling Circle ◽  
Isothermal Nucleic Acid Amplification

The early detection of invasive fungal infection (IFD) is significant in order to decrease mortality in susceptible patients. There is, therefore, a need for sensitive and specific fungal species detection assays in a clinical laboratory for early targeted therapy. The isothermal amplification method may be useful for the screening of fungal isolates, especially in resource-poor settings. Therefore, our aim was to review the isothermal nucleic acid amplification methods and their applications in fungal pathogen detection. Out of 50 reported studies, 28, 12, 6, 2, and 2 studies used the isothermal-based assays of a loop-mediated isothermal amplification (LAMP), nucleic acid sequence-based amplification (NASBA), rolling circle amplification (RCA), multiple displacement amplification (MDA) and polymerase Spiral Reaction (PSR), respectively. Thirty-two studies used clinical samples, 18 pure culture, and four environmental samples. The diagnostic accuracy of isothermal nucleic acid amplification testing for pathogenic fungal was reported as high (sensitivity 0.89–1.0 and specificity 0.63–1.0) in all studies irrespective of the sample tested. Although the isothermal-based assays showed high sensitivity and specificity in reported studies, it is still poorer than that of PCR assays. However, improving the assay to make it simpler, more effective, and inexpensive compared with newer PCR methods are still needed.

A novel method to control carryover contamination in isothermal nucleic acid amplification

2017 ◽  
Vol 53 (77) ◽  
pp. 10696-10699 ◽  
Author(s):  
Cuiping Ma ◽  
Fuxin Wang ◽  
Xiudan Wang ◽  
Lingzhi Han ◽  
Hao Jing ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Restriction Endonuclease ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Isothermal Nucleic Acid Amplification ◽  
Recognition Sites ◽  
Novel Method

We developed a novel method to control carryover contamination in loop-mediated isothermal amplification (LAMP) by primer engineering to carry recognition sites for a restriction endonuclease, providing a robust ability to eliminate carryover contaminants.

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