Nucleic acid sequence-based amplification

Compton, Jean

doi:10.1038/350091a0

Cited by 1,195 publications

(668 citation statements)

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“…Real-time PCR assay has been useful to study the transmission and development of this viral infection in juvenile [56]. Another useful method is the nucleic acid sequence based amplification (NASBA) [27] which is an isothermal method for nucleic acid amplification that is particularly suited to RNA targets [68]. The method amplifies a target-specific product through oligonucleotide primers and the co-ordinated activity of 3 enzymes: reverse transcriptase, RNase H, and T7 RNA polymerase.…”

Section: Microscopymentioning

confidence: 99%

Betanodavirus of Marine and Freshwater Fish: Distribution, Genomic Organization, Diagnosis and Control Measures

et al. 2012

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Section: Microscopymentioning

confidence: 99%

Betanodavirus of Marine and Freshwater Fish: Distribution, Genomic Organization, Diagnosis and Control Measures

et al. 2012

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“…Thermal cycling of the PCR technique imposes instrumental constraints, limiting the technique to a laboratory setting, and dual-labelled fluorescent probes, such as Taqman probes 12 , are usually needed to determine the specificity of amplification. Therefore, isothermal amplification of RNA, such as nucleic acid sequence-based amplification [13][14][15] , rolling-cycle amplification 16,17 and loopmediated isothermal amplification 18,19 have emerged as alternative amplification techniques. As the above mentioned reactions can be preceded at a constant temperature, there is no need of specialized instruments for RNA detection, and in addition, they have potential for 'on-site' testing.…”

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confidence: 99%

“…Two kinds of catalytic DNA sequences, RNA-cleaving DNAzyme (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23) 26,27 and G-quadruplex horseradish peroxidase-mimicking DNAzyme (PW17) [28][29][30] were applied in our research. The 10-23 RNAcleaving DNAzyme has been reported to cleave any purinepyrimidine (RY) junction under simulated physiological conditions 26 ; hence, it was applied in our strategy for the cleaving of target RNA molecule to initiate the following exponential amplification.…”

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Cleavage-based signal amplification of RNA

2013

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RNA detection has become an integral part of current biomedical research. Up to now, the reverse transcription-PCR has been the most practical method to detect mRNA targets. However, RNA detection by reverse transcription-PCR requires sophisticated equipment and it is highly sensitive to contamination with genomic DNA. Here we report a new isothermal reaction to simultaneously amplify and detect RNA, based on cleavage by DNAzyme and signal amplification. Cleavage-based signal amplification of RNA cannot be contaminated by genomic DNA and is suitable for the detection of both mRNA and microRNA targets, with high specificity and sensitivity. Moreover, the detection results can be reported in a colorimetric or real-time fluorometric way for different detection purposes.

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“…Nucleic acid sequence based amplification (NASBA) is a technology developed in the early 90s to amplify nucleic acids without the use of a thermal cycler [9]. It is most often used to obtain many copies of RNA starting from a few RNA molecules.…”

Section: Nucleic Acid Sequence Based Amplification (Nasba)mentioning

confidence: 99%

Potentials and limitations of molecular diagnostic methods in food safety

Lauri

Mariani

2008

Genes Nutr

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Molecular methods allow the detection of pathogen nucleic acids (DNA and RNA) and, therefore, the detection of contamination in food is carried out with high selectivity and rapidity. In the last 2 decades molecular methods have accompanied traditional diagnostic methods in routine pathogen detection, and might replace them in the upcoming future. In this review the implementation in diagnostics of four of the most used molecular techniques (PCR, NASBA, microarray, LDR) are described and compared, highlighting advantages and limitations of each of them. Drawbacks of molecular methods with regard to traditional ones and the difficulties encountered in pathogen detection from food or clinical specimen are also discussed. Moreover, criteria for the choice of the target sequence for a secure detection and classification of pathogens and possible developments in molecular diagnostics are also proposed.

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Nucleic acid sequence-based amplification

Abstract: Nucleic acid sequence-based amplification (NASBA) is a primer-dependent technology that can be used for the continuous amplification of nucleic acids in a single mixture at one temperature.

Cited by 1,195 publications

References 0 publications

Betanodavirus of Marine and Freshwater Fish: Distribution, Genomic Organization, Diagnosis and Control Measures

Betanodavirus of Marine and Freshwater Fish: Distribution, Genomic Organization, Diagnosis and Control Measures

Cleavage-based signal amplification of RNA

Potentials and limitations of molecular diagnostic methods in food safety

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