Isothermal Nucleic Acid Amplification Techniques and Their Use in Bioanalysis

Bodulev, Oleg L.; Sakharov, I Iu

doi:10.1134/s0006297920020030

Cited by 98 publications

(59 citation statements)

References 122 publications

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“…As a result, isothermal nucleic acid amplification has emerged as an alternative detection method for SARS-CoV-2 viral RNA from clinical samples [ 130 ]. In general, isothermal amplification techniques increase the analytical signal by increasing the target nucleic acid concentration through enzymatic activities at a fixed temperature, and simultaneously detecting the signal with colorimetric or fluorescence indicators [ 131 ]. Changes in color, fluorescence level, or turbidity indicate the presence of SARS-CoV-2 RNA or DNA [ 131 ].…”

Section: Isothermal Nucleic Acid Amplification Methodsmentioning

confidence: 99%

“…In general, isothermal amplification techniques increase the analytical signal by increasing the target nucleic acid concentration through enzymatic activities at a fixed temperature, and simultaneously detecting the signal with colorimetric or fluorescence indicators [ 131 ]. Changes in color, fluorescence level, or turbidity indicate the presence of SARS-CoV-2 RNA or DNA [ 131 ]. Therefore, unlike RT-qPCR, isothermal amplification methods do not require thermal cycling instruments or specialized technicians for disease diagnosis [ 132 ].…”

Section: Isothermal Nucleic Acid Amplification Methodsmentioning

confidence: 99%

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Nucleic Acid Testing of SARS-CoV-2

Yoo

Kim

2021

IJMS

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The coronavirus disease 2019 (COVID-19) has caused a large global outbreak. It is accordingly important to develop accurate and rapid diagnostic methods. The polymerase chain reaction (PCR)-based method including reverse transcription-polymerase chain reaction (RT-PCR) is the most widely used assay for the detection of SARS-CoV-2 RNA. Along with the RT-PCR method, digital PCR has emerged as a powerful tool to quantify nucleic acid of the virus with high accuracy and sensitivity. Non-PCR based techniques such as reverse transcription loop-mediated isothermal amplification (RT-LAMP) and reverse transcription recombinase polymerase amplification (RT-RPA) are considered to be rapid and simple nucleic acid detection methods and were reviewed in this paper. Non-conventional molecular diagnostic methods including next-generation sequencing (NGS), CRISPR-based assays and nanotechnology are improving the accuracy and sensitivity of COVID-19 diagnosis. In this review, we also focus on standardization of SARS-CoV-2 nucleic acid testing and the activity of the National Metrology Institutes (NMIs) and highlight resources such as reference materials (RM) that provide the values of specified properties. Finally, we summarize the useful resources for convenient COVID-19 molecular diagnostics.

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Section: Isothermal Nucleic Acid Amplification Methodsmentioning

confidence: 99%

Section: Isothermal Nucleic Acid Amplification Methodsmentioning

confidence: 99%

Nucleic Acid Testing of SARS-CoV-2

Yoo

Kim

2021

IJMS

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“…Isothermal amplification methods have been widely used for species identification of pathogenic fungi in recent years ( He et al, 2019 ; Qiu et al, 2019 ; Bodulev and Sakharov, 2020 ). Among them, loop-mediated isothermal amplification (LAMP) is the most widely used one, which has received increasing attention ( Lee, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

Rapid and Visual Identification of Chlorophyllum molybdites With Loop-Mediated Isothermal Amplification Method

et al. 2021

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Chlorophyllum molybdites is a kind of common poisonous mushroom in China that is widely distributed in different areas. Food poisoning caused by accidentally eating C. molybdites has become more frequent in recent years. In 2019, there were 55 food poisoning incidents caused by eating this mushroom in China. Mushroom poisoning continues to be a common health issue of global concern. When mushroom poisoning occurs, an effective, simple, and rapid detection method is required for accurate clinical treatment or forensic analysis. For the first time, we established a loop-mediated isothermal amplification (LAMP) assay for the visual detection of C. molybdites. A set of specific LAMP primers was designed, and the specificity was confirmed against 43 different mushroom species. The LAMP method could detect as low as 1 pg of genomic DNA. Boiled mushrooms and artificial gastric-digested mushroom samples were prepared to test the applicability of the method, and the results showed that as low as 1% C. molybdites in boiled and digested samples could be successfully detected. The LAMP method can also be completed within 45 min, and the reaction results could be directly observed based on a color change under daylight by the naked eye. Therefore, the LAMP assay established in this study provides an accurate, sensitive, rapid, and low-cost method for the detection of C. molybdites.

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“…In recent years, many other amplification techniques have been developed to improve PCR in terms of sensitivity, affordability, and rapidity [7]. Loop-mediated isothermal amplification (LAMP) is one of the most widely used isothermal methods, detecting bacteria, DNA viruses, and, recently, parasites [8][9][10][11][12].…”

Section: Introductionmentioning

confidence: 99%

One-Day Molecular Detection of Salmonella and Campylobacter in Chicken Meat: A Pilot Study

Zendrini

Carta

Filipello

et al. 2021

Foods

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Salmonella and Campylobacter ssp. are bacterial pathogens responsible for most foodborne infections in EU countries. Poultry serves as a reservoir for these pathogens, and its important role in the meat industry makes it essential to develop a rapid detection assay able to provide results in one day. Indeed, the rapid identification of foodborne pathogens is an important instrument for the monitoring and prevention of epidemic outbreaks. To date, Salmonella and Campylobacter screening is mainly conducted through molecular methods (PCR or real-time PCR) performed after 18–24 h long enrichments. In this study, we evaluated short enrichments (0, 2, 4, and 6 h) combined with a colorimetric loop-mediated isothermal AMPlification (LAMP) or real-time PCR to detect Salmonella and Campylobacter in poultry meat contaminated at different concentration levels (101, 103, and 105 CFU/g). Our results show that real-time PCR allows the detection of Salmonella and Campylobacter, even after shorter enrichment times than prescribed by ISO references; particularly, it detected Salmonella down to 101 CFU/g since T0 and Campylobacter from 103 CFU/g since T0. Detection with LAMP was comparable to real-time PCR without the requirement of a thermal cycler and with shorter execution times. These characteristics make colorimetric LAMP a valid alternative when one-day results are needed, improving the timely identification of positive meat batches, even in the absence of specialized instrumentation.

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Isothermal Nucleic Acid Amplification Techniques and Their Use in Bioanalysis

Cited by 98 publications

References 122 publications

Nucleic Acid Testing of SARS-CoV-2

Nucleic Acid Testing of SARS-CoV-2

Rapid and Visual Identification of Chlorophyllum molybdites With Loop-Mediated Isothermal Amplification Method

One-Day Molecular Detection of Salmonella and Campylobacter in Chicken Meat: A Pilot Study

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