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Genomics and Bionanotechnology
Published in Anil Kumar Anal, Bionanotechnology, 2018
DNA amplification is the process to copy number of sequences of DNA. Polymerase chain reaction (PCR) invented by Kary B. Mullis in 1980s is the first DNA amplification technique designed initially to study DNA in small quantities (Mullis 1990). With the advancements in PCR, it is possible to isolate and detect target DNA sequence, mutation or polymorphisms, paternity of child, genetic diseases, study human genome, and match DNA in forensic science. Different nucleic acid amplification techniques based on PCR have been modified for applications in molecular medicine. For instance, ligase chain reaction (LCR) is used for gene mutation characterization and detection of infectious diseases; nucleic acid sequence-based amplification (NASBA) and related transcription-mediated amplification are used for infection diagnosis such as human immunodeficiency virus (HIV), hepatitis, and branched DNA-based detection for diagnostic purpose (Sorscher 1997).
Lateral Flow Assays
Published in Sibel A. Ozkan, Bengi Uslu, Mustafa Kemal Sezgintürk, Biosensors, 2023
Kamil Żukowski, Marcin Drozd, Robert Ziółkowski, Mariusz Pietrzak, Katarzyna Tokarska, Adam Nowiński, Elżbieta Malinowska
In real life applications the NALFA test are used for detection of DNA fragments amplified during PCR (45–47). It should be stressed, that in the case of classic, three temperature-based PCR reactions, the nucleic acids amplification step still is limited only to well-equipped laboratories with skilled staff. However, the reports dedicated to the use of LFA tests as detection element in the case isothermal nucleic acids amplification methods (48–56), show the technological approach which truly could be appropriate for the point-of-need, thus fulfilling the ASSURED criteria (affordable, sensitive, specific, user-friendly, rapid and robust, equipment-free and deliverable to end-users). That is mainly because theoretically for the nucleic acid amplification or its detection no energy source is needed and the whole assay could be performed by unskilled persons. Importantly, this approach still offers high selectivity, low cost and satisfactory time of the analysis. Among different methods of isothermal DNA amplification the nucleic acid sequence based amplification (NASBA), transcription mediated amplification (TMA), self-sustained sequence replication (3SR), helicase dependent amplification (HAD), loop mediated isothermal amplification (LAMP) or recombinase polymerase amplification (RPA) should be mentioned. The latter seems to offer the most user friendly approach (57). This is because it requires conventionally designed primers and leads to exponential amplification with no need for pretreatment of DNA sample. Moreover, the reactions are sensitive, specific and rapid and the whole system works at constant low temperature (optimum in the range between 37 and 40o C) (57). It is a reason for multiple examples of LFAs dedicated to determination/detection of RPA products, in a form of both NALFIA and NALFT (35, 57–60) (see Fig. 10.7).
Developing mitochondrial DNA field-compatible tests
Published in Critical Reviews in Environmental Science and Technology, 2022
Bidhan C. Dhar, Christina E. Roche, Jay F. Levine
Several alternative nucleic acid based isothermal amplification methods have been developed (Kim & Easley, 2011). Among these are loop mediated isothermal amplification (LAMP; Notomi et al. 2000), recombinase polymerase amplification (RPA; Piepenburg et al., 2006), nucleic acid sequence-based amplification (NASBA, also known as transcription mediated amplification; Compton, 1991), and rolling circle amplification (RCA; Dean et al., 2001). Although these methods can differ considerably, all share some common characteristics (Figure 1).