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
Detection of Mycobacterium ulcerans by the loop mediated isothermal amplification method.
Anthony Ablordey,Diana Amissah,I.F. Aboagye,Ben Hatano,Toshio Yamazaki,Tetsutaro Sata,Koichi Ishikawa,Harutaka Katano +7 more
TL;DR: The LAMP method can be used as a simple and rapid test for the detection of M. ulcerans in clinical specimens, however, obtaining purified DNA, as well as generating isothermal conditions, remains a major challenge for the use of the Lamp method under field conditions.
62
Application of DNA-based diagnostics in detection of schistosomal DNA in early infection and after drug treatment
TL;DR: The data presented here indicate that LAMP is suitable for the detection of early infection in the groups primarily infected with Schistosoma japonicum, such as migrants, travellers, military personnel and the younger age groups, but is less suitable for evaluation of the efficacy of chemotherapy in the early stages.
Analytical Performance of Four Polymerase Chain Reaction (PCR) and Real Time PCR (qPCR) Assays for the Detection of Six Leishmania Species DNA in Colombia
Cielo León,Cielo León,Marina Muñoz,Carolina Hernández,Martha S. Ayala,Carolina Flórez,Aníbal A. Teherán,Juan Ricardo Cubides,Juan David Ramírez +8 more
TL;DR: The present study revealed that the 18S marker presented the best performance in terms of analytical sensitivity and specificity for the qPCR in the species tested (species circulating in Colombia), and recommends to explore the analytical and diagnostic performance in future studies using a broader number of species across America.
62
Sensitive and rapid detection of Clonorchis sinensis infection in fish by loop-mediated isothermal amplification (LAMP)
X. Q. Cai,Min-Jun Xu,Y. H. Wang,D. Y. Qiu,G. X. Liu,A. Lin,J. D. Tang,Renli Zhang,Xing-Quang Zhu,Xing-Quang Zhu +9 more
TL;DR: The LAMP assay was proved to be 100 times more sensitive than a conventional polymerase chain reaction for detection of C. sinensis in fish, which has important implications for the effective control of human clonorchiasis.
62
Advances in the Diagnosis of Foot-and-Mouth Disease.
TL;DR: In the current review, enzyme-linked immunosorbent assay (ELISA)-based methods used in FMD diagnosis are extensively reviewed, particularly the sandwich, liquid-phase blocking, and solid-phase competition ELISA.
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