Digital droplet LAMP as a microfluidic app on standard laboratory devices

Schuler, Friedrich; Siber, Clara; Hin, Sebastian; Wadle, Simon; Paust, Nils; Zengerle, Roland; Stetten, Felix von

doi:10.1039/c6ay00600k

Cited by 48 publications

(39 citation statements)

References 21 publications

(32 reference statements)

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“…We summarize the chamber size and number, together with the consequent sensitivity and dynamic range of the dLAMP in Table 1 . In most studies, the volume of the microchambers is less than 10 nL . The chamber number is around 1000; further increase in the compartment number will not increase the resolution of dLAMP, as confirmed by previous simulation results .…”

Section: Current Dlamp Techniquessupporting

confidence: 71%

“…By increasing the vibration frequency and the flow rate, the size of the resultant droplets can be decreased. In addition to the flow focusing method and the XiE method, LAMP reagents can also be emulsified using a commercial Dropchip powered by a centrifuge as shown in Figure a3, which produces droplets with a CV of 1.4% . Consequently, the flow focusing, XiE and the centrifuge methods are all capable of generating droplets with a narrow size distribution, ensuring the accuracy of the nucleic acid quantification of the dLAMP technique.…”

Section: Current Dlamp Techniquesmentioning

confidence: 99%

“…a1–a3) Typical devices to emulsify LAMP mixtures. (a1 is reproduced with permission, Copyright 2019, American Chemical Society; a2 is reproduced with permission, Copyright 2016, American Chemical Society; a3 reproduced with permission, Copyright 2016, Royal Society of Chemistry.) b1, b2) The multilayer approach for preventing evaporation during heating.…”

Section: Current Dlamp Techniquesmentioning

confidence: 99%

“…The compartment number is a crucial parameter in droplet microfluidic dLAMP, as it can affect the sensitivity, the dynamic range, and the resolution of nucleic acid amplification inside droplets. Currently, varying numbers of compartments have been applied, including 250, 1000, 5000, and 10 000 droplets per µL, which can provide different sensitivities and dynamic ranges, as shown in Table 3 . The sensitivity bellow 10 copies per µL can be achieved in the test with a partition number of 20 000 droplets per test or more; this is comparable to the sensitivity of dPCR.…”

Section: Current Dlamp Techniquesmentioning

confidence: 99%

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Droplet and Microchamber‐Based Digital Loop‐Mediated Isothermal Amplification (dLAMP)

Yuan

Chao

Shum

2020

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Digital loop‐mediated isothermal amplification (dLAMP) refers to compartmentalizing nucleic acids and LAMP reagents into a large number of individual partitions, such as microchambers and droplets. This compartmentalization enables dLAMP to be an excellent platform to quantify the absolute number of the target nucleic acids. Owing to its low requirement for instrumentation complexity, high specificity, and strong tolerance to inhibitors in the nucleic acid samples, dLAMP has been recognized as a simple and accurate technique to quantify pathogenic nucleic acid. Herein, the general process of dLAMP techniques is summarized, the current dLAMP techniques are categorized, and a comprehensive discussion on different types of dLAMP techniques is presented. Also, the challenges of the current dLAMP are illustrated together with the possible strategies to address these challenges. In the end, the future directions of the dLAMP developments, including multitarget detection, multisample detection, and processing nucleic acid extraction are outlined. With recently significant advances in dLAMP, this technology has the potential to see more widespread use beyond the laboratory in the future.

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Section: Current Dlamp Techniquessupporting

confidence: 71%

Section: Current Dlamp Techniquesmentioning

confidence: 99%

Section: Current Dlamp Techniquesmentioning

confidence: 99%

Section: Current Dlamp Techniquesmentioning

confidence: 99%

See 2 more Smart Citations

Droplet and Microchamber‐Based Digital Loop‐Mediated Isothermal Amplification (dLAMP)

Yuan

Chao

Shum

2020

Small

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“…Another method was introduced using disposable polymeric DropChip, generating the droplets by centrifugal force [58]. Once the droplet generation was completed, the chip was transferred to a thermal cycler to perform LAMP in z1 h and the amplification results were monitored by a microarray scanner.…”

Section: Digital Lampmentioning

confidence: 99%

LAMP-on-a-chip: Revising microfluidic platforms for loop-mediated DNA amplification

Zhang

Fohlerová

et al. 2019

TrAC Trends in Analytical Chemistry

171

113

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a b s t r a c tNucleic acid amplification for the detection of infectious diseases, food pathogens, or assessment of genetic disorders require a laboratory setting with specialized equipment and technical expertise. Isothermal deoxyribonucleic acid amplification methods, such as loop-mediated isothermal amplification (LAMP), exhibit characteristics ideal for point-of-care (POC) applications, since their instrumentation is simpler in comparison with the standard method of polymerase chain reaction. Other key advantages of LAMP are robustness and the production of pyrophosphate in the presence of the target gene, enabling to detect the reaction products using the naked eye. Polymerase inhibitors, presented in clinical samples, do not affect the amplification process, making LAMP suitable for a simple sample-to-answer diagnostic systems with simplified sample preparation. In this review, we discuss the trends in miniaturized LAMP techniques, such as microfluidic, paper-based, and digital with their advantages and disadvantages, especially for POC applications alongside our opinion of the future development of miniaturized LAMP.

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Droplet Encoding‐Pairing Enabled Multiplexed Digital Loop‐Mediated Isothermal Amplification for Simultaneous Quantitative Detection of Multiple Pathogens

et al. 2023

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Despite the advantages of digital nucleic acid analysis (DNAA) in terms of sensitivity, precision, and resolution, current DNAA methods commonly suffer a limitation in multiplexing capacity. To address this issue, a droplet encoding‐pairing enabled DNAA multiplexing strategy is developed, wherein unique tricolor combinations are deployed to index individual primer droplets. The template droplets and primer droplets are sequentially introduced into a microfluidic chip with a calabash‐shaped microwell array and are pairwise trapped and merged in the microwells. Pre‐merging and post‐amplification image analysis with a machine learning algorithm is used to identify, enumerate, and address the droplets. By incorporating the amplification signals with droplet encoding information, simultaneous quantitative detection of multiple targets is achieved. This strategy allows for the establishment of flexible multiplexed DNAA by simply adjusting the primer droplet library. Its flexibility is demonstrated by establishing two multiplexed (8‐plex) droplet digital loop‐mediated isothermal amplification (mddLAMP) assays for individually detecting lower respiratory tract infection and urinary tract infection causative pathogens. Clinical sample analysis shows that the microbial detection outcomes of the mddLAMP assays are consistent with those of the conventional assay. This DNAA multiplexing strategy can achieve flexible high‐order multiplexing on demand, making it a desirable tool for high‐content pathogen detection.

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Digital droplet LAMP as a microfluidic app on standard laboratory devices

Abstract: Digital droplet LAMP is performed on a disposable chip (DropChip) with the size of a microscope slide using only standard laboratory devices.

Cited by 48 publications

References 21 publications

Droplet and Microchamber‐Based Digital Loop‐Mediated Isothermal Amplification (dLAMP)

Droplet and Microchamber‐Based Digital Loop‐Mediated Isothermal Amplification (dLAMP)

LAMP-on-a-chip: Revising microfluidic platforms for loop-mediated DNA amplification

Droplet Encoding‐Pairing Enabled Multiplexed Digital Loop‐Mediated Isothermal Amplification for Simultaneous Quantitative Detection of Multiple Pathogens

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