Development of a fluorescent distance-based paper device using loop-mediated isothermal amplification to detect <i>Escherichia coli</i> in urine

Saengsawang, Natkrittaya; Ruang-areerate, Toon; Kesakomol, Piyanate; Thita, Thanyapit; Mungthin, Mathirut; Dungchai, Wijitar

doi:10.1039/d0an01306d

Cited by 19 publications

(37 citation statements)

References 48 publications

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“…Its simplicity and low operating cost make this technique an important asset when resources are limited. Hence, even if it is still increasingly used in the clinical detection of human pathogens (e.g., 6 – 9 ), assays have also been developed in various other fields, such as pathogen detection in crops 10 – 12 , in aquaculture 13 , in food safety 14 , 15 or in food market control for species identification 16 .…”

Section: Introductionmentioning

confidence: 99%

eDNA-based detection of the invasive crayfish Pacifastacus leniusculus in streams with a LAMP assay using dependent replicates to gain higher sensitivity

Porco

Hermant

Purnomo

et al. 2022

Sci Rep

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LAMP assays are becoming increasingly popular in the field of invasive species detection but are still underused in eDNA-based monitoring. Here, we propose a LAMP assay designed to detect the North American crayfish species Pacifastacus leniusculus in water samples from streams. The presence of P. leniusculus was detected through this new LAMP assay in all but one of the nine sites sampled. No correlation was found between ddPCR absolute concentration measurements and the number of LAMP-positive technical replicates. However, we showed that using dependent technical replicates could significantly enhance the detection sensitivity of the LAMP assay. Applied to other assays, it could improve sensitivity and thus allow for a more efficient use of eDNA-based LAMP assays for invasive species detection in aquatic ecosystems.

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Section: Introductionmentioning

confidence: 99%

eDNA-based detection of the invasive crayfish Pacifastacus leniusculus in streams with a LAMP assay using dependent replicates to gain higher sensitivity

Porco

Hermant

Purnomo

et al. 2022

Sci Rep

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“…In particular, without the necessity of using expensive thermal circulators, gel electrophoresis and UV detection equipment, LAMP is low-cost. Currently, quantitative LAMP has been developed in broad spectrums including cheap semi-quantified E. coli LAMP using paper-based on costly LAMP chips (Rolando et al 2019 ; Saengsawang et al 2021 ). It is suitable for the rapid detection of a pathogenic microorganism on the spot, the popularization of field and grass-roots in wartime (Mori et al 2013 ).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of the real-time fluorescence loop-mediated isothermal amplification assay for the detection of Ureaplasma urealyticum

Shen

Lao³

et al. 2022

AMB Expr

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Ureaplasma urealyticum (UU) is commonly present in human reproductive tract, which frequently leads to genital tract infection. Hence, there is an urgent need to develop a rapid detection method for UU. In our study, a real-time fluorescence loop-mediated isothermal amplification (LAMP) assay was developed and evaluated for the detection of UU. Two primers were specifically designed based on the highly conserved regions of ureaseB genes. The reaction was carried out for 60 min in a constant temperature system using Bst DNA polymerase, and the process was monitored by real-time fluorescence signal, while polymerase chain reaction (PCR) was performed simultaneously. In real-time fluorescence LAMP reaction system, positive result was only obtained for UU among 9 bacterial strains, with detection sensitivity of 42 pg/μL (4.2 × 105 CFU/mL), and all 16 clinical samples of UU could be detected. In conclusion, real-time fluorescence LAMP is a simple, sensitive, specific and effective method compared with conventional PCR, which shows great promise in the rapid detection of UU.

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“…Contrastingly, LAMP requires basic equipment, whereas local trained health care workers could perform the assay according to its simple operating procedure. To overcome the limitations of LAMP in the field sites, simplified methods have been successfully modified and adapted to the assays such as selective colorimetric and fluorescent dyes 11 , 13 , 23 , 44 , lyophilized form 45 , paper-based devices 46 and dye capsules 15 , 18 . In the meantime, ongoing development of the new approaches encourages the use of LAMP for point-of-care (POC) in low resource settings such as health services as well as field sites; not only for screening but also to confirm diagnostic level.…”

Section: Discussionmentioning

confidence: 99%

“…The one-step SYBR safe and AuNP-LAMP assay are suitable for multipurpose Leishmania detection, for example, direct evaluation or screening in field environments using closed tube SYBR safe method to reduce contamination prone 13 , 19 or, re-evaluation and verification using post hybridization of AuNPs-probe method to control and assure the first detection 26 , 40 . In addition, the remaining LAMP products could be further quantified by a distance-based paper device 46 . These applications could improve and additionally simplify the closed tube SYBR safe dye LAMP assay to empower asymptomatic leishmaniasis surveillance in HIV patients.…”

Section: Discussionmentioning

confidence: 99%

Development of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patients

Ruang-areerate

Sukphattanaudomchoke

Thita

et al. 2021

Sci Rep

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Asymptomatic leishmaniasis cases have continuously increased, especially among patients with HIV who are at risk to develop further symptoms of cutaneous and visceral leishmaniasis. Thus, early diagnosis using a simple, sensitive and reliable diagnostic assay is important because populations at risk mostly reside in rural communities where laboratory equipment is limited. In this study, the highly sensitive and selective determination of Leishmania infection in asymptomatic HIV patients was achieved using dual indicators (SYBR safe and gold-nanoparticle probe; AuNP-probe) in one-step LAMP method based on basic instruments. The assay can be simply evaluated under the naked eye due to clear interpretation of fluorescent emission of LAMP-SYBR safe dye-complex and colorimetric precipitate of specific AuNP-probes. The sensitivities and specificities of fluorescent SYBR safe dye and AuNP-probe indicators were equal, which were as high as 94.1 and 97.1%, respectively. Additionally, detection limits were 102 parasites/mL (0.0147 ng/µL), ten times more sensitivity than other related studies. To empower leishmaniasis surveillance, this inexpensive one-step SYBR safe and AuNP-LAMP assay is reliably fast and simple for field diagnostics to point-of-care settings, which can be set up in all levels of health care facilities including resource limited areas, especially in low to middle income countries.

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Development of a fluorescent distance-based paper device using loop-mediated isothermal amplification to detect Escherichia coli in urine

Abstract: The highly sensitive and selective determination of Escherichia coli (E. coli) in urine using SYBRTM Safe loop-mediated Isothermal Amplification (LAMP) method with a distance-based paper device was developed. The new...

Cited by 19 publications

References 48 publications

eDNA-based detection of the invasive crayfish Pacifastacus leniusculus in streams with a LAMP assay using dependent replicates to gain higher sensitivity

eDNA-based detection of the invasive crayfish Pacifastacus leniusculus in streams with a LAMP assay using dependent replicates to gain higher sensitivity

Evaluation of the real-time fluorescence loop-mediated isothermal amplification assay for the detection of Ureaplasma urealyticum

Development of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patients

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