Competitive annealing mediated isothermal amplification of nucleic acids

The Analyst ◽  
2018 ◽  
Vol 143 (3) ◽  
pp. 639-642 ◽  
Author(s):  
Rui Mao ◽  
Lifei Qi ◽  
Jianjun Li ◽  
Ming Sun ◽  
Zhuo Wang ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
High Specificity ◽  
Isothermal Amplification ◽  
Amplification Method

A novel nucleic acid isothermal amplification method with high specificity, efficiency and rapidity was developed.

Loop-mediated isothermal amplification technique: principle, development and wide application in food safety

2020 ◽  
Vol 12 (46) ◽  
pp. 5551-5561
Author(s):  
Tianzeng Huang ◽  
Linzhi Li ◽  
Xing Liu ◽  
Qi Chen ◽  
Xueen Fang ◽  
...  
Keyword(s):  
Food Safety ◽  
Gene Amplification ◽  
Foodborne Pathogens ◽  
High Specificity ◽  
Isothermal Amplification ◽  
Isothermal Conditions ◽  
Loop Mediated Isothermal Amplification ◽  
Novel Gene ◽  
Amplification Method ◽  
Amplification Technique

LAMP is a relatively novel gene amplification method under isothermal conditions with rapidity, and high specificity. It is widely applied in the field of food safety, such as in the detection of foodborne pathogens, GM, OP pesticides and so on

scholarly journals IsoPCR: An Analytically Sensitive, Nested, Multiplex Nucleic Acid Amplification Method

2013 ◽  
Vol 59 (2) ◽  
pp. 436-439 ◽  
Author(s):  
Martin Jensen Søe ◽  
Mikkel Rohde ◽  
Jens Mikkelsen ◽  
Peter Warthoe
Keyword(s):  
Nucleic Acid ◽  
Candida Glabrata ◽  
Simultaneous Detection ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Qpcr Assay ◽  
Nucleic Acid Amplification ◽  
Molecular Diagnostic ◽  
Nucleic Acid Detection ◽  
Amplification Method

BACKGROUND Nucleic acid tests that can simultaneously detect multiple targets with high sensitivity, specificity, and speed are highly desirable. To meet this need, we developed a new approach we call the isoPCR method. METHODS The isoPCR method is a 2-stage nested-like nucleic acid amplification method that combines a single multiplex preamplification PCR with subsequent distinct detection of specific targets by use of isothermal amplification. We compared isoPCR to nested quantitative PCR (qPCR), loop-mediated isothermal amplification (LAMP), and nested LAMP (PCR followed by LAMP), for detection of DNA from Candida glabrata. We evaluated the method's multiplex capability for detecting low copy numbers of pathogens commonly involved in sepsis. RESULTS IsoPCR provided detection of 1 copy of Candida glabrata, an LOD that was 5-fold lower than a nested qPCR assay (5 copies), while the amplification time was simultaneously halved. Similarly, the LOD for isoPCR was lower than that for a LAMP assay (1000 copies) and a nested LAMP assay (5 copies). IsoPCR required recognition of 6 regions for detection, thereby providing a theoretically higher specificity compared to nested qPCR (4 regions). The isoPCR multiplexing capability was demonstrated by simultaneous detection of 4 pathogens with individual LODs of 10 copies or fewer. Furthermore, the specificity of isoPCR was demonstrated by successful pathogen detection from samples with more than 1 pathogen present. CONCLUSIONS IsoPCR provides a molecular diagnostic tool for multiplex nucleic acid detection, with an LOD down to 1 copy, high theoretical specificity, and halving of the amplification time compared to a nested qPCR assay.

scholarly journals Loop-Mediated Isothermal Amplification (LAMP) for Detection of Various Organisms: A Review

2017 ◽  
Vol 11 (2) ◽  
pp. 20-27
Author(s):  
Arifa Akram
Keyword(s):  
Nucleic Acid ◽  
High Specificity ◽  
Disease Diagnosis ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Lamp Method ◽  
Isolation And Identification ◽  
Strand Displacement Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Prophylactic Measures

Disease diagnosis is important for implementation of proper therapeutic and prophylactic measures. Traditionally, disease diagnosis was depends upon isolation and identification of the causative organisms. This was followed by serology and after that molecular method. Molecular tests are valuable when early diagnosis is important. For this purpose, nucleic acid amplification (PCR, nucleic acid sequence-based amplification, self-sustained sequence replication, strand displacement amplification) is one of the most valuable tools not only for the diagnosis of infectious diseases but also used in advanced level research. The Loop-Mediated Isothermal Amplification (LAMP) is a unique nucleic acid amplification technique for diagnosis of various pathogens introduced at 2000 by Notomi and his colleagues which is simple, easy, rapid and cost effective when compared to PCR due to its high specificity, sensitivity, and rapidity. It uses a set of six primers and a DNA polymerase with stranddisplacement activity. Major advantage of LAMP method is its cost-effectiveness as it can be done simply by using waterbath or heating block. Bangladesh J Med Microbiol 2017; 11 (2): 20-27

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.

Shortening distance of forward and reverse primers for nucleic acid isothermal amplification

2014 ◽  
Vol 395 (6) ◽  
pp. 679-684
Author(s):  
Qu Haitao ◽  
Zhang Wenchao ◽  
Zhang Xiaohui ◽  
Wang Xiujun ◽  
Li Sulong
Keyword(s):  
Nucleic Acid ◽  
Reaction Times ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Amplification Efficiency ◽  
Specific Method ◽  
Detection Techniques ◽  
Amplification Method ◽  
Reverse Primer ◽  
Short Time

Abstract Existent nucleic acid isothermal detection techniques for clinical diseases are difficult to promote greatly due to limitations in such aspects as methodology, costs of detection, amplification efficiency and conditions for operation. There is therefore an urgent need for a new isothermal amplification method with the characteristics of high accuracy, easy operation, short time of detection and low costs. We have devised a new method of nucleic acid isothermal amplification using Bst DNA polymerase under isothermal conditions (60–65°C). We call this method of amplification by shortening the distance between forward and reverse primers for nucleic acid isothermal amplification SDAMP. The results demonstrated that this technique is highly sensitive, specific and has short reaction times (40–60 min). Results of sequencing show that the products of SDAMP amplification are mainly polymers formed by series connection of monomers formed through linkage of forward primer and complementary sequences in reverse primer via a few bases. The method is different from current methods of nucleic acid amplification. Our study shows, however, that it is a specific method of nucleic acid isothermal amplification depending on interactions between primers and DNA template.

NAIL: Nucleic Acid detection using Isotachophoresis and Loop-mediated isothermal amplification

Lab on a Chip ◽  
2015 ◽  
Vol 15 (7) ◽  
pp. 1697-1707 ◽  
Author(s):  
Mark D. Borysiak ◽  
Kevin W. Kimura ◽  
Jonathan D. Posner
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Mobile Phone ◽  
Isothermal Amplification ◽  
Nucleic Acid Detection ◽  
Complex Matrices ◽  
Loop Mediated Isothermal Amplification

The NAIL device integrates isotachophoresis and loop-mediated isothermal amplification (LAMP) with mobile phone detection to extract, amplify, and detect nucleic acids from complex matrices in less than one hour.

scholarly journals Three-Way Junction-Induced Isothermal Amplification with High Signal-to-Background Ratio for Detection of Pathogenic Bacteria

Sensors ◽  
2021 ◽  
Vol 21 (12) ◽  
pp. 4132
Author(s):  
Jung Ho Kim ◽  
Seokjoon Kim ◽  
Sung Hyun Hwang ◽  
Tae Hwi Yoon ◽  
Jung Soo Park ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Pathogenic Bacteria ◽  
High Selectivity ◽  
Isothermal Amplification ◽  
Rna Aptamers ◽  
High Signal ◽  
Fluorescence Signal ◽  
Enhanced Fluorescence ◽  
Target Nucleic Acid

The consumption of water and food contaminated by pathogens is a major cause of numerous diseases and deaths globally. To control pathogen contamination and reduce the risk of illness, a system is required that can quickly detect and monitor target pathogens. We developed a simple and reproducible strategy, termed three-way junction (3WJ)-induced transcription amplification, to detect target nucleic acids by rationally combining 3WJ-induced isothermal amplification with a light-up RNA aptamer. In principle, the presence of the target nucleic acid generates a large number of light-up RNA aptamers (Spinach aptamers) through strand displacement and transcription amplification for 2 h at 37 °C. The resulting Spinach RNA aptamers specifically bind to fluorogens such as 3,5-difluoro-4-hydroxybenzylidene imidazolinone and emit a highly enhanced fluorescence signal, which is clearly distinguished from the signal emitted in the absence of the target nucleic acid. With the proposed strategy, concentrations of target nucleic acids selected from the genome of Salmonellaenterica serovar Typhi (S. Typhi) were quantitatively determined with high selectivity. In addition, the practical applicability of the method was demonstrated by performing spike-and-recovery experiments with S. Typhi in human serum.

Flap endonuclease-initiated enzymatic repairing amplification for ultrasensitive detection of target nucleic acids

Nanoscale ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 3633-3638 ◽  
Author(s):  
Hyowon Jang ◽  
Chang Yeol Lee ◽  
Seoyoung Lee ◽  
Ki Soo Park ◽  
Hyun Gyu Park
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Nucleic Acid Amplification ◽  
Ultrasensitive Detection ◽  
Isothermal Nucleic Acid Amplification ◽  
Amplification Method

A new isothermal nucleic acid amplification method termed FERA (Flap endonuclease-initiated Enzymatic Repairing Amplification) is developed for the ultrasensitive detection of target nucleic acids.

scholarly journals PATH-27. A RAPID COLORIMETRIC PEPTIDE NUCLEIC ACID LOOP-MEDIATED ISOTHERMAL AMPLIFICATION METHOD FOR THE DETECTION OF THE IDH1 MUTATION IN GLIOBLASTOMA

2017 ◽  
Vol 19 (suppl_6) ◽  
pp. vi176-vi176 ◽  
Author(s):  
Edward Raack ◽  
Matthew Jennings ◽  
Robert Belton ◽  
Johnathan Lawrence ◽  
Christopher McMahon ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Peptide Nucleic Acid ◽  
Idh1 Mutation ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Method
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