Integrated Glass Microdevice for Nucleic Acid Purification, Loop-Mediated Isothermal Amplification, and Online Detection

2011 ◽  
Vol 83 (9) ◽  
pp. 3336-3342 ◽  
Author(s):  
Qingqing Wu ◽  
Wei Jin ◽  
Chao Zhou ◽  
Sihai Han ◽  
Wenxiu Yang ◽  
...  
2017 ◽  
Vol 11 (2) ◽  
pp. 20-27
Author(s):  
Arifa Akram

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


Animals ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 76
Author(s):  
Faiz Padzil ◽  
Abdul Razak Mariatulqabtiah ◽  
Wen Siang Tan ◽  
Kok Lian Ho ◽  
Nurulfiza Mat Isa ◽  
...  

Over the years, development of molecular diagnostics has evolved significantly in the detection of pathogens within humans and their surroundings. Researchers have discovered new species and strains of viruses, while mitigating the viral infections that occur, owing to the accessibility of nucleic acid screening methods such as polymerase chain reaction (PCR), qualitative (real-time) polymerase chain reaction (qPCR) and reverse-transcription qPCR (RT-qPCR). While such molecular detection methods are widely utilized as the benchmark, the invention of isothermal amplifications has also emerged as a reliable tool to improvise on-field diagnosis without dependence on thermocyclers. Among the established isothermal amplification technologies are loop-mediated isothermal amplification (LAMP), recombinant polymerase amplification (RPA), strand displacement activity (SDA), nucleic acid sequence-based amplification (NASBA), helicase-dependent amplification (HDA) and rolling circle amplification (RCA). This review highlights the past research on and future prospects of LAMP, its principles and applications as a promising point-of-care diagnostic method against avian viruses.


2018 ◽  
Vol 268 ◽  
pp. 255-263 ◽  
Author(s):  
Padmavathy Bakthavathsalam ◽  
Guillaume Longatte ◽  
Slade O. Jensen ◽  
Mike Manefield ◽  
J. Justin Gooding

Viruses ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 699 ◽  
Author(s):  
Mahapatra ◽  
Howson ◽  
Fowler ◽  
Batten ◽  
Flannery ◽  
...  

Peste des petits ruminants (PPR) is a disease of small ruminants caused by peste des petits ruminants virus (PPRV), and is endemic in Asia, the Middle East and Africa. Effective control combines the application of early warning systems, accurate laboratory diagnosis and reporting, animal movement restrictions, suitable vaccination and surveillance programs, and the coordination of all these measures by efficient veterinary services. Molecular assays, including conventional reverse transcription-polymerase chain reaction (RT-PCR) and real-time RT-PCR (RT-qPCR) have improved the sensitivity and rapidity of diagnosing PPR. However, currently these assays are only performed within laboratory settings; therefore, the development of field diagnostics for PPR would improve the fast implementation of control policies, particularly when PPR has been targeted to be eradicated by 2030. Loop-mediated isothermal amplification (LAMP) assays are simple to use, rapid, and have sensitivity and specificity within the range of RT-qPCR; and can be performed in the field using disposable consumables and portable equipment. This study describes the development of a novel RT-LAMP assay for the detection of PPRV nucleic acid by targeting the N-protein gene. The RT-LAMP assay was evaluated using cell culture propagated PPRVs, field samples from clinically infected animals and samples from experimentally infected animals encompassing all four lineages (I-IV) of PPRV. The test displayed 100% concordance with RT-qPCR when considering an RT-qPCR cut-off value of CT >40. Further, the RT-LAMP assay was evaluated using experimental and outbreak samples without prior RNA extraction making it more time and cost-effective. This assay provides a solution for a pen-side, rapid and inexpensive PPR diagnostic for use in the field in nascent PPR eradication programme.


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