Rapid quantification of fecal indicator bacteria in water using the most probable number - loop-mediated isothermal amplification (MPN-LAMP) approach on a polymethyl methacrylate (PMMA) microchip

Fu, Jing; Chiang, Elaine L. C.; Medriano, Carl Angelo; Li, Liyan; Bae, Sung Woo

doi:10.1016/j.watres.2021.117172

Cited by 17 publications

(13 citation statements)

References 49 publications

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“…167−170 To avoid the application of expensive and complicated equipment, Fu et al investigated a rapid on-chip most probable number (MPN)−LAMP approach to determine the target genes of fecal indicator bacteria (FIB) in water samples. 171 ously. 172 This system consisted of a microchip and a portable device for temperature control and results visualization (Figure 4).…”

Section: ■ Microscale Systems "See" In Cellsmentioning

confidence: 99%

“…LAMP involves four to six primers and a strand-displacing DNA polymerase to promote the amplification process at a constant temperature. − LAMP provides a quick and selective technology for detecting foodborne microorganisms . In addition to controlling microbial contamination in food, it is important to explore fast and sensitive methods for detecting bacteria in water samples. − To avoid the application of expensive and complicated equipment, Fu et al investigated a rapid on-chip most probable number (MPN)–LAMP approach to determine the target genes of fecal indicator bacteria (FIB) in water samples . A microchip platform combined with LAMP was developed to detect Streptococcus pneumoniae ( S. pneumoniae ) and Mycoplasma pneumoniae ( M. pneumoniae ) simultaneously .…”

Section: Microscale Systems “See” In Bacteriamentioning

confidence: 99%

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What the Microscale Systems “See” In Biological Assemblies: Cells and Viruses?

et al. 2021

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Section: ■ Microscale Systems "See" In Cellsmentioning

confidence: 99%

Section: Microscale Systems “See” In Bacteriamentioning

confidence: 99%

What the Microscale Systems “See” In Biological Assemblies: Cells and Viruses?

et al. 2021

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“…The assay achieved a LOD of 2.5 × 10 2 copies/reaction in 30 min. In another study, Fu et al [ 122 ] developed a LAMP assay using the most probable number (MNP) for the quantification of E. coli and Enterococcus spp. in a PMMA chip.…”

Section: Lamp-based Point-of-care Biosensorsmentioning

confidence: 99%

LAMP-Based Point-of-Care Biosensors for Rapid Pathogen Detection

2022

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Seeking optimized infectious pathogen detection tools is of primary importance to lessen the spread of infections, allowing prompt medical attention for the infected. Among nucleic-acid-based sensing techniques, loop-mediated isothermal amplification is a promising method, as it provides rapid, sensitive, and specific detection of microbial and viral pathogens and has enormous potential to transform current point-of-care molecular diagnostics. In this review, the advances in LAMP-based point-of-care diagnostics assays developed during the past few years for rapid and sensitive detection of infectious pathogens are outlined. The numerous detection methods of LAMP-based biosensors are discussed in an end-point and real-time manner with ideal examples. We also summarize the trends in LAMP-on-a-chip modalities, such as classical microfluidic, paper-based, and digital LAMP, with their merits and limitations. Finally, we provide our opinion on the future improvement of on-chip LAMP methods. This review serves as an overview of recent breakthroughs in the LAMP approach and their potential for use in the diagnosis of existing and emerging diseases.

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“… 20 , 21 To address these challenges, isothermal NAA methods, such as loop-mediated isothermal amplification (LAMP), are used for environmental quantification of other microbial pathogens including Zika virus, 22 astrovirus, 23 MS2, 24 Escherichia coli , and Enterococcus spp. 25 , 26 RT-LAMP quantification has higher tolerances to inhibitors and shorter amplification times (e.g., 30 min) compared to RT-qPCR. 24 , 27 , 28 Huang et al used a portable in-gel LAMP platform for the sensitive detection of MS2 coliphage in wastewater, while RT-qPCR failed to produce a positive result.…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, RT-qPCR does not produce absolute quantification. In addition, RT-qPCR targeting SARS-CoV-2 is sensitive to inhibitors that are present in wastewater, leading to false-negative results. , To address these challenges, isothermal NAA methods, such as loop-mediated isothermal amplification (LAMP), are used for environmental quantification of other microbial pathogens including Zika virus, astrovirus, MS2, Escherichia coli, and Enterococcus spp. , RT-LAMP quantification has higher tolerances to inhibitors and shorter amplification times (e.g., 30 min) compared to RT-qPCR. ,, Huang et al used a portable in-gel LAMP platform for the sensitive detection of MS2 coliphage in wastewater, while RT-qPCR failed to produce a positive result . RT-LAMP has been used in portable SARS-CoV-2 detection platforms for use on clinical samples. , However, isothermal methods have yet to be developed for detecting SARS-CoV-2 in environmental water samples.…”

Section: Introductionmentioning

confidence: 99%

Membrane-Based In-Gel Loop-Mediated Isothermal Amplification (mgLAMP) System for SARS-CoV-2 Quantification in Environmental Waters

Zhu

et al. 2021

Environ. Sci. Technol.

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Since the COVID-19 pandemic is expected to become endemic, quantification of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in ambient waters is critical for environmental surveillance and for early detection of outbreaks. Herein, we report the development of a membrane-based in-gel loop-mediated isothermal amplification (mgLAMP) system that is designed for the rapid point-of-use quantification of SARS-CoV-2 particles in environmental waters. The mgLAMP system integrates the viral concentration, in-assay viral lysis, and on-membrane hydrogel-based RT-LAMP quantification using enhanced fluorescence detection with a target-specific probe. With a sample-to-result time of less than 1 h, mgLAMP successfully detected SARS-CoV-2 below 0.96 copies/mL in Milli-Q water. In surface water, the lowest detected SARS-CoV-2 concentration was 93 copies/mL for mgLAMP, while the reverse transcription quantitative polymerase chain reaction (RT-qPCR) with optimal pretreatment was inhibited at 930 copies/mL. A 3D-printed portable device is designed to integrate heated incubation and fluorescence illumination for the simultaneous analysis of nine mgLAMP assays. Smartphone-based imaging and machine learning-based image processing are used for the interpretation of results. In this report, we demonstrate that mgLAMP is a promising method for large-scale environmental surveillance of SARS-CoV-2 without the need for specialized equipment, highly trained personnel, and labor-intensive procedures.

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Rapid quantification of fecal indicator bacteria in water using the most probable number - loop-mediated isothermal amplification (MPN-LAMP) approach on a polymethyl methacrylate (PMMA) microchip

Cited by 17 publications

References 49 publications

What the Microscale Systems “See” In Biological Assemblies: Cells and Viruses?

What the Microscale Systems “See” In Biological Assemblies: Cells and Viruses?

LAMP-Based Point-of-Care Biosensors for Rapid Pathogen Detection

Membrane-Based In-Gel Loop-Mediated Isothermal Amplification (mgLAMP) System for SARS-CoV-2 Quantification in Environmental Waters

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