Loop-mediated isothermal amplification of DNA
Tsugunori Notomi,Hiroto Okayama,Harumi Otawara-shi Masubuchi,Toshihiro Yonekawa,Keiko Watanabe,Nobuyuki Amino,Tetsu Hase +6 more
TL;DR: A novel method that amplifies DNA with high specificity, efficiency and rapidity under isothermal conditions that employs a DNA polymerase and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA.
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Abstract: We have developed a novel method, termed loop-mediated isothermal amplification (LAMP), that amplifies DNA with high specificity, efficiency and rapidity under isothermal conditions. This method employs a DNA polymerase and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA. An inner primer containing sequences of the sense and antisense strands of the target DNA initiates LAMP. The following strand displacement DNA synthesis primed by an outer primer releases a single-stranded DNA. This serves as template for DNA synthesis primed by the second inner and outer primers that hybridize to the other end of the target, which produces a stem–loop DNA structure. In subsequent LAMP cycling one inner primer hybridizes to the loop on the product and initiates displacement DNA synthesis, yielding the original stem–loop DNA and a new stem–loop DNA with a stem twice as long. The cycling reaction continues with accumulation of 109 copies of target in less than an hour. The final products are stem–loop DNAs with several inverted repeats of the target and cauliflower-like structures with multiple loops formed by annealing between alternately inverted repeats of the target in the same strand. Because LAMP recognizes the target by six distinct sequences initially and by four distinct sequences afterwards, it is expected to amplify the target sequence with high selectivity.
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Citations
Visual Detection of High-Risk Human Papillomavirus Genotypes 16, 18, 45, 52, and 58 by Loop-Mediated Isothermal Amplification with Hydroxynaphthol Blue Dye
TL;DR: This qualitative and colorimetric LAMP assay has potential usefulness for the rapid screening of HPV genotype 16, 18, 45, 52, and 58 infections, especially in resource-limited hospitals or rural clinics of provincial and municipal regions in China.
62
Rapid detection of multiple respiratory viruses based on microfluidic isothermal amplification and a real-time colorimetric method
Wang Ruili,Rongtao Zhao,Yang Li,Kong Wen,Guo Xudong,Yi Yang,Feng Wu,Wanying Liu,Song Hongbin,Hao Rongzhang +9 more
TL;DR: The developed method provides an effective alternative for rapid point-of-care screening for viruses that cause respiratory disease syndromes and further aids in accurate and timely detection to control and prevent the spread of respiratory diseases caused by such pathogens.
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Detection of phytoplasma by loop-mediated isothermal amplification of DNA (LAMP).
TL;DR: LAMP is validates for routine diagnosis of Napier stunt and other closely related phytoplasmas and was 20-fold more sensitive than nested PCR.
62
Specific detection of reverse transcription-loop-mediated isothermal amplification amplicons for Taura syndrome virus by colorimetric dot-blot hybridization.
TL;DR: A field diagnosis system based on isothermal reverse transcription-loop-mediated amplification for shrimp Taura syndrome virus (TSV), placing emphasis on specific and simple detection of the LAMP amplicons, and a one-step guanidinium thiocyanate homogenization method to provide RNA extraction efficiency comparable to that of the TRIzol Reagent for RT-LAMP.
62
Development of diagnostics for aquaculture: challenges and opportunities
TL;DR: This paper looks at some of the opportunities and challenges for the development of rapid diagnostics for aquaculture.
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