Nucleic Acids Hybridization: Potentials and Limitations

Buzdin, Anton

doi:10.1007/978-1-4020-6040-3_1

Cited by 2 publications

(2 citation statements)

References 71 publications

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“…By contrast, nucleic acid hybridizations have the advantage of detecting a specific gene or nucleic-acid sequence, so gene expression is not a prerequisite. Unfortunately, these methods are tedious and more sophisticated and complex than traditional methodologies [19][20][21]. PCR and qPCR assays have limitations in pathogen identification because of the need for specialized laboratory equipment and reagents, as well as proper training and technical expertise, which are often unavailable in poorly-resourced laboratory and field conditions [22][23][24].…”

Section: Introductionmentioning

confidence: 99%

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

Waliullah

Ling

Cieniewicz

et al. 2020

IJMS

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A loop-mediated isothermal amplification (LAMP) assay was developed for simple, rapid and efficient detection of Cucurbit leaf crumple virus (CuLCrV), one of the most important begomoviruses that infects cucurbits worldwide. A set of six specific primers targeting a total 240 nt sequence regions in the DNA A of CuLCrV were designed and synthesized for detection of CuLCrV from infected leaf tissues using real-time LAMP amplification with the Genie® III system, which was further confirmed by gel electrophoresis and SYBR™ Green I DNA staining for visual observation. The optimum reaction temperature and time were determined, and no cross-reactivity was seen with other begomoviruses. The LAMP assay could amplify CuLCrV from a mixed virus assay. The sensitivity assay demonstrated that the LAMP reaction was more sensitive than conventional PCR, but less sensitive than qPCR. However, it was simpler and faster than the other assays evaluated. The LAMP assay also amplified CuLCrV-infected symptomatic and asymptomatic samples more efficiently than PCR. Successful LAMP amplification was observed in mixed virus-infected field samples. This simple, rapid, and sensitive method has the capacity to detect CuLCrV in samples collected in the field and is therefore suitable for early detection of the disease to reduce the risk of epidemics.

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Section: Introductionmentioning

confidence: 99%

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

Waliullah

Ling

Cieniewicz

et al. 2020

IJMS

View full text Add to dashboard Cite

show abstract

“…In hybridization assays, a labeled oligonucleotide probe that is complementary to the genomic sequence of interest is used to demonstrate the presence of a particular sequence in the sample. However, these traditional molecular methods have disadvantages of being labor-intensive, cumbersome, and requiring large amounts of nucleic acids, so they are not routinely used in diagnostic laboratories [ 104 , 105 ].…”

Section: Detection Of Sars-cov-2mentioning

confidence: 99%

Wastewater-Based Epidemiology as an Early Warning System for the Spreading of SARS-CoV-2 and Its Mutations in the Population

Martı́n

Gál

Špalková

et al. 2021

IJERPH

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New methodologies based on the principle of “sewage epidemiology” have been successfully applied before in the detection of illegal drugs. The study describes the idea of early detection of a virus, e.g., SARS-CoV-2, in wastewater in order to focus on the area of virus occurrence and supplement the results obtained from clinical examination. By monitoring temporal variation in viral loads in wastewater in combination with other analysis, a virus outbreak can be detected and its spread can be suppressed early. The use of biosensors for virus detection also seems to be an interesting application. Biosensors are highly sensitive, selective, and portable and offer a way for fast analysis. This manuscript provides an overview of the current situation in the area of wastewater analysis, including genetic sequencing regarding viral detection and the technological solution of an early warning system for wastewater monitoring based on biosensors.

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Nucleic Acids Hybridization: Potentials and Limitations

Cited by 2 publications

References 71 publications

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

Wastewater-Based Epidemiology as an Early Warning System for the Spreading of SARS-CoV-2 and Its Mutations in the Population

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