Simultaneous elimination of carryover contamination and detection of DNA with uracil-DNA-glycosylase-supplemented loop-mediated isothermal amplification (UDG-LAMP)

Hsieh, Kuangwen; Mage, Peter L.; Csordas, Andrew T.; Eisenstein, Michael; Soh, H. Tom

doi:10.1039/c4cc00540f

Cited by 163 publications

(142 citation statements)

References 24 publications

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“…Furthermore, as indicated, the results can be inspected visually, so the method has considerable potential for application in low-resource countries and is highly cost-effective. However, one important limitation is that the DNA amplification mechanism involved in the LAMP technique itself could lead to carryover contamination, giving rise to false-positive results (230,233). Also, other possible problems, such as difficulties in optimization and the limitations of multiplexing associated with the use of increasing numbers of LAMP primers, need to be considered in applying this approach (234,235).…”

Section: Lampmentioning

confidence: 99%

Advances in the Diagnosis of Human Schistosomiasis

et al. 2015

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SUMMARY Schistosomiasis is a major neglected tropical disease that afflicts more than 240 million people, including many children and young adults, in the tropics and subtropics. The disease is characterized by chronic infections with significant residual morbidity and is of considerable public health importance, with substantial socioeconomic impacts on impoverished communities. Morbidity reduction and eventual elimination through integrated intervention measures are the focuses of current schistosomiasis control programs. Precise diagnosis of schistosome infections, in both mammalian and snail intermediate hosts, will play a pivotal role in achieving these goals. Nevertheless, despite extensive efforts over several decades, the search for sensitive and specific diagnostics for schistosomiasis is ongoing. Here we review the area, paying attention to earlier approaches but emphasizing recent developments in the search for new diagnostics for schistosomiasis with practical applications in the research laboratory, the clinic, and the field. Careful and rigorous validation of these assays and their cost-effectiveness will be needed, however, prior to their adoption in support of policy decisions for national public health programs aimed at the control and elimination of schistosomiasis.

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Section: Lampmentioning

confidence: 99%

Advances in the Diagnosis of Human Schistosomiasis

et al. 2015

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“…The powerful amplification mechanism of LAMP, which is beneficial for having highly sensitive assays, also renders it highly susceptible to carry over contamination, where amplified DNA products from previous LAMP reactions become templates for reamplification (Hsieh, Mage, Csordas, Eisenstein, & Soh, 2014;Tomita et al, 2008). Consequently, one of our goals was to find one single incubation temperature that would work accurately for both CHD-W and CHD-Z primer sets so that separate reactions could be run simultaneously on the same thermoblock, requiring less equipment and time overall.…”

Section: Discussionmentioning

confidence: 99%

Rapid sex determination of a wild passerine species using loop‐mediated isothermal amplification (LAMP)

Koch

Blohm‐Sievers

Liedvogel

2019

Ecology and Evolution

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Many bird species are sexually monomorphic and cannot be sexed based on phenotypic traits. Rapid sex determination is often a necessary component of avian studies focusing on behavior, ecology, evolution, and conservation. While PCR‐based methods are the most common technique for molecularly sexing birds in the laboratory, a simpler, faster, and cheaper method has emerged, which can be used in the laboratory, but importantly also in the field. Herein, we used loop‐mediated isothermal amplification (LAMP) for rapid sex determination of blood samples from juvenile European blackcaps, Sylvia atricapilla, sampled in the wild. We designed LAMP primers unique to S. atricapilla based on the sex chromosome‐specific gene, chromo‐helicase‐DNA‐binding protein (CHD), optimized the primers for laboratory and field application, and then used them to test a subset of wild‐caught juvenile blackcaps of unknown gender at the time of capture. Sex determination results were fast and accurate. The advantages of this technique are that it allows researchers to identify the sex of individual birds within hours of sampling and eliminates the need for direct access to a laboratory if implemented at a remote field site. This work adds to the increasing list of available LAMP primers for different bird species and is a new addition within the Passeriformes order.

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“…One major drawback of LAMP is its high risk of carryover contamination which often leads to false-positive results in negative controls (Hsieh et al 2014;Karthik et al 2014). This is due to the extremely high efficiency of the LAMP reaction.…”

Section: Limitations Of Lampmentioning

confidence: 99%

Loop-mediated isothermal amplification (LAMP): a versatile technique for detection of micro-organisms

et al. 2018

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Simultaneous elimination of carryover contamination and detection of DNA with uracil-DNA-glycosylase-supplemented loop-mediated isothermal amplification (UDG-LAMP)

Abstract: We report a one-pot, closed-vessel enzymatic assay that eliminates carryover contamination while preserving robust DNA amplification in loop-mediated isothermal amplification (LAMP), providing reliable and rapid detection of target DNA in contaminated samples.

Cited by 163 publications

References 24 publications

Advances in the Diagnosis of Human Schistosomiasis

Advances in the Diagnosis of Human Schistosomiasis

Rapid sex determination of a wild passerine species using loop‐mediated isothermal amplification (LAMP)

Loop-mediated isothermal amplification (LAMP): a versatile technique for detection of micro-organisms

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