Pullulan reduces the non-specific amplification of loop-mediated isothermal amplification (LAMP)

Gao, Xueqin; Sun, Bingqi; Guan, Yifu

doi:10.1007/s00216-018-1552-2

Cited by 43 publications

(18 citation statements)

References 34 publications

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“…This device operates by detecting viral nucleic acids such as the reference standard (RT-PCR) and isothermal amplification tests. RT-LAMP amplification unfortunately has issues such as: non-specific amplification that can be a problem in later steps as a result of “carry-over contamination”, “product cross contamination” [ 28 , 29 , 30 ], or primer hybridization [ 31 ].…”

Section: Discussionmentioning

confidence: 99%

Analytical Evaluation of Visby Medical RT-PCR Portable Device for Rapid Detection of SARS-CoV-2

et al. 2021

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Extended community testing constitutes one of the main strategic pillars in controlling the COVID-19 pandemic. Reverse transcription PCR (RT-PCR) targeting the SARS-CoV-2 genome on nasopharyngeal swab samples is currently the reference test. While displaying excellent analytical sensitivity and specificity, this test is costly, often requires a substantial turnaround time, and, more importantly, is subject to reagent and other material shortages. To complement this technology, rapid antigen tests have been developed and made available worldwide, allowing cheap, quick, and decentralized SARS-CoV-2 testing. The main drawback of these tests is the reduced sensitivity when RT-PCR is the gold standard. In this study, we evaluate Visby an innovative, portable, easy-to-use RT-PCR point-of-care (POC) diagnostic device. Our retrospective analysis shows that overall, compared to the Cobas 6800 RT-qPCR assay (Roche), this RT-PCR POC technology detects SARS-CoV-2 RNA with 95% sensitivity (95%CI = 86.3–99%) and 100% specificity (95% CI = 80.5–100%). For samples with cycle-threshold values below 31, we observed 100% sensitivity (95% CI = 66.4–100%). While showing an analytical sensitivity slightly below that of a standard RT-qPCR system, the evaluated Visby RT-PCR POC device may prove to be an interesting diagnostic alternative in the COVID-19 pandemic, potentially combining the practical advantages of rapid antigen tests and the robust analytical performances of nucleic acid detection systems.

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Section: Discussionmentioning

confidence: 99%

Analytical Evaluation of Visby Medical RT-PCR Portable Device for Rapid Detection of SARS-CoV-2

et al. 2021

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“…Hence, LAMP can be used as a reli able method for the diagnostics of Plasmodium species. However, in another study, LAMP based assay produced some false positives [15]. LAMP was successfully used for identification of Salmonella in food products in situ and detection of stxA2 (Shiga toxin 2 subunit A) in Escherichia coli O157:H7 cells [15a, 15b].…”

Section: Amplification Techniques That Increase Analytical Signal By mentioning

confidence: 99%

Isothermal Nucleic Acid Amplification Techniques and Their Use in Bioanalysis

Bodulev

Sakharov

2020

Biochemistry Moscow

100

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“…One of the limitations of colorimetric LAMP-based methods based on the phenol red color shift is that some degree of color change exists in samples containing no template (see ref [29]). This has been associated to the occurrence of nonspecific amplification induced by the interaction among the LAMP primers [24,34], which often serve as templates for off-target polymerizations. In general, nonspecific amplification is believed to be the main reason for false positives in LAMP colorimetric reactions.…”

Section: Lf-qlamp In Saliva Samples: Discrimination Between Positive and Negative Samplesmentioning

confidence: 99%

“…LAMP-based methods also use from four to six primer sets, and the design of LAMP primer sets is nontrivial as the primers may interact among themselves during amplification. This can promote nonspecific amplifications that may also result in false positives [23,24].…”

Section: Introductionmentioning

confidence: 99%

Portable and Label-Free Quantitative Loop-Mediated Isothermal Amplification (LF-qLamp) for Reliable COVID-19 Diagnostics in Three Minutes of Reaction Time: Arduino-Based Detection System Assisted by a pH Microelectrode

et al. 2021

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Loop-mediated isothermal amplification (LAMP) has been recently studied as an alternative method for cost-effective diagnostics in the context of the current COVID-19 pandemic. Recent reports document that LAMP-based diagnostic methods have a comparable sensitivity and specificity to that of RT-qPCR. We report the use of a portable Arduino-based LAMP-based amplification system assisted by pH microelectrodes for the accurate and reliable diagnosis of SARS-CoV-2 during the first 3 min of the amplification reaction. We show that this simple system enables a straightforward discrimination between samples containing or not containing artificial SARS-CoV-2 genetic material in the range of 10 to 10,000 copies per 50 µL of reaction mix. We also spiked saliva samples with SARS-CoV-2 synthetic material and corroborated that the LAMP reaction can be successfully monitored in real time using microelectrodes in saliva samples as well. These results may have profound implications for the design of real-time and portable quantitative systems for the reliable detection of viral pathogens including SARS-CoV-2.

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Pullulan reduces the non-specific amplification of loop-mediated isothermal amplification (LAMP)

Cited by 43 publications

References 34 publications

Analytical Evaluation of Visby Medical RT-PCR Portable Device for Rapid Detection of SARS-CoV-2

Analytical Evaluation of Visby Medical RT-PCR Portable Device for Rapid Detection of SARS-CoV-2

Isothermal Nucleic Acid Amplification Techniques and Their Use in Bioanalysis

Portable and Label-Free Quantitative Loop-Mediated Isothermal Amplification (LF-qLamp) for Reliable COVID-19 Diagnostics in Three Minutes of Reaction Time: Arduino-Based Detection System Assisted by a pH Microelectrode

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