Challenges and perspectives in the application of isothermal DNA amplification methods for food and water analysis

Martzy, Roland; Kolm, Claudia; Krska, Rudolf; Mach, Robert L.; Farnleitner, Andreas H.; Reischer, Georg H.

doi:10.1007/s00216-018-1553-1

Cited by 54 publications

(44 citation statements)

References 38 publications

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“…In contrast, isothermal methods require only single, constant temperature throughout the reaction. A simple heat block or even water bath could be used to maintain the reaction temperature needed 35 . Furthermore, an isothermal reaction can reduce the overall reaction time by eliminating the time necessary for temperature adjustment.…”

Section: Discussionmentioning

confidence: 99%

Sensitive fluorescence detection of SARS-CoV-2 RNA in clinical samples via one-pot isothermal ligation and transcription

et al. 2020

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The control of viral outbreaks requires nucleic acid diagnostic tests that are sensitive, simple and fast. Here, we report a highly sensitive and specific one-pot assay for the fluorescence-based detection of RNA from pathogens. The assay, which can be performed within 30-50 min of incubation time and can reach a limit of detection of 0.1-attomolar RNA concentration, relies on a sustained isothermal reaction cascade producing an RNA aptamer that binds to a fluorogenic dye. The RNA aptamer is transcribed by the T7 RNA polymerase from the ligation product of a promoter DNA probe and a reporter DNA probe that hybridize with the target single-stranded RNA sequence via the SplintR ligase (a Chlorella virus DNA ligase). In 40 nasopharyngeal SARS-CoV-2 samples, the assay reached positive and negative predictive values of 95 and 100%, respectively. We also show that the assay can rapidly detect a range of viral and bacterial RNAs.

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Section: Discussionmentioning

confidence: 99%

Sensitive fluorescence detection of SARS-CoV-2 RNA in clinical samples via one-pot isothermal ligation and transcription

et al. 2020

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“…Isothermal amplification methods have been developed (e.g. LAMP and RPA) that are faster than PCR and can be operated at constant temperature, eliminating the need for sophisticated equipment like thermocyclers ( Martzy et al, 2019 ). However, these new advantages come with tradeoffs in terms of sensitivity and specificity.…”

Section: Introductionmentioning

confidence: 99%

Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities

Dongen

Berendsen

Steenbergen

et al. 2020

Biosensors and Bioelectronics

284

191

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With the trend of moving molecular tests from clinical laboratories to on-site testing, there is a need for nucleic acid based diagnostic tools combining the sensitivity, specificity and flexibility of established diagnostics with the ease, cost effectiveness and speed of isothermal amplification and detection methods. A promising new nucleic acid detection method is Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated nuclease (Cas)-based sensing. In this method Cas effector proteins are used as highly specific sequence recognition elements that can be combined with many different read-out methods for on-site point-of-care testing. This review covers the technical aspects of integrating CRISPR/Cas technology in miniaturized sensors for analysis on-site. We start with a short introduction to CRISPR/Cas systems and the different effector proteins and continue with reviewing the recent developments of integrating CRISPR sensing in miniaturized sensors for point-of-care applications. Finally, we discuss the challenges of point-of-care CRISPR sensing and describe future research perspectives.

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“…Isothermal nucleic acid amplification is a technique that is used to amplify nucleic acids at constant temperature avoiding the complex requirement of the regular PCR that needs changing multiple temperatures in each cycle [ 70 , 71 ].…”

Section: Current Detection Methods For Sars-cov-2mentioning

confidence: 99%

Diagnostic techniques for COVID-19 and new developments

Sheikhzadeh

Eissa

Ismail

et al. 2020

Talanta

129

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COVID-19 pandemic is a serious global health issue today due to the rapid human to human transmission of SARS-CoV-2, a new type of coronavirus that causes fatal pneumonia. SARS -CoV-2 has a faster rate of transmission than other coronaviruses such as SARS and MERS and until now there are no approved specific drugs or vaccines for treatment. Thus, early diagnosis is crucial to prevent the extensive spread of the disease. The reverse transcription-polymerase chain reaction (RT-PCR) is the most routinely used method until now to detect SARS-CoV-2 infections. However, several other faster and accurate assays are being developed for the diagnosis of COVID-19 aiming to control the spread of infection through the identification of patients and immediate isolation. In this review, we will discuss the various detection methods of the SARS-CoV-2 virus including the recent developments in immunological assays, amplification techniques as well as biosensors.

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Challenges and perspectives in the application of isothermal DNA amplification methods for food and water analysis

Cited by 54 publications

References 38 publications

Sensitive fluorescence detection of SARS-CoV-2 RNA in clinical samples via one-pot isothermal ligation and transcription

Sensitive fluorescence detection of SARS-CoV-2 RNA in clinical samples via one-pot isothermal ligation and transcription

Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities

Diagnostic techniques for COVID-19 and new developments

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