Multiplex loop-mediated isothermal amplification-based lateral flow dipstick for simultaneous detection of 3 food-borne pathogens in powdered infant formula

Jiang, Yujun; Chen, Sihan; Zhao, Yueming; Yang, Xinyan; Fu, Shiqian; McKillip, John L.; Fox, Edward M.; Man, Chaoxin

doi:10.3168/jds.2019-17538

Cited by 42 publications

(24 citation statements)

References 36 publications

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“…There are conflicting reports regarding the role of different types of sugars in maintaining protein’s stability. Various studies reported trehalose as an efficient stabilizer because of its higher glass transition temperature (Tg) as compared to sucrose [ 22 , 23 , 24 ]. On contrary, there are studies that have demonstrated the employment of sucrose for preserving protein structure [ 17 , 25 , 26 ].…”

Section: Resultsmentioning

confidence: 99%

Optimizing Effective Parameters to Enhance the Sensitivity of Vertical Flow Assay for Detection of Escherichia coli

Kaur

Eltzov

2022

Biosensors

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Vertical flow immunoassays (VFIAs) are considered potential point-of-care testing (POCT) devices compared to lateral flow assays due to their ability to analyze a comparatively large sample volume and ease of multiplexing. However, VFIA devices are limited by low analytical sensitivity when coupled with a visual colorimetric signal. Herein, we carefully analyzed key parameters that accounted for the proper functionality of VFIA that can be modified to enhance the overall sensitivity of VFIA. In particular, we focused on improving the stability of conjugate pads impregnated with capture antibodies, maintaining a controlled flow rate to ensure higher analyte reactivity with capture antibodies, and enhancing the absorption efficiency. The results showed that air-drying of conjugate pads in the presence of 5% (w/v) lactose significantly improved the stability of antibodies during long-term storage. Integration of dissolvable polyvinyl alcohol (PVA) membrane of optimal concentration as a time-barrier film into the sensor delayed the flow of samples, thereby increasing the biorecognition interaction time between immunoreagents for the formation of immuno-complexes, which in turn led to higher sensitivity of the assay. Furthermore, the employment of an absorbent pad with higher water holding capacity significantly reduced the non-specific binding of immunocomplexes, thereby reducing the possibility of false-negative results.

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Section: Resultsmentioning

confidence: 99%

Optimizing Effective Parameters to Enhance the Sensitivity of Vertical Flow Assay for Detection of Escherichia coli

Kaur

Eltzov

2022

Biosensors

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“…Since the introduction of LAMP technology over 20 years ago, it has been applied in many settings, from point-of-care rapid clinical testing for coronaviruses ( Augustine et al, 2020 ) to multiplex lateral-flow dipstick tests for simultaneous rapid detection of multiple pathogens in foods ( Lalle et al, 2018 ; Jiang et al, 2020 ). Here, we applied a previously developed LAMP test to the detection of Giardia in leafy greens, using a comparatively simple approach that could be readily employed in most food testing laboratories for routine surveillance, using existing equipment and expertise and commercially available reagents.…”

Section: Discussionmentioning

confidence: 99%

Optimization and validation of a loop-mediated isothermal amplification (LAMP) assay for detection of Giardia duodenalis in leafy greens

Lalonde

Xie

Oakley

et al. 2021

Food and Waterborne Parasitology

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Giardia duodenalis is one of the most common food and water-borne intestinal parasites of humans and animals worldwide. Fresh, ready-to-eat produce such as leafy greens and salad mixes are considered potential transmission vehicles for Giardia infection in humans. Therefore, a specific, sensitive, and reliable method for Giardia detection in leafy greens is needed. We optimized washing procedures for the recovery of Giardia cysts from leafy greens and adapted and validated an existing EF1α LAMP assay for the detection of Giardia DNA to support routine diagnostic surveillance and disease outbreak investigations. Four leafy green types (35 ± 1 g) were spiked with 100 Giardia cysts and we compared washing by shaking with 1 M glycine ( n = 20) or 0.1% Alconox ( n = 20). DNA was extracted from washes, tested by LAMP and melt curve analysis, and time to positive (TTP) values compared. The detection limit was determined by spiking 10 ( n = 40) Giardia cysts onto these same types of leafy greens and processing as above with 0.1% Alconox. Method robustness was assessed by subjecting spring mix ( n = 45 total) to aging (1, 3 or 7 days) and washes to aging and freezing conditions prior to testing. Assay repeatability and specificity were evaluated, and an artificial positive control (APC) distinguishable by melt temperature (Tm) from DNA of Giardia spiked on leafy greens was designed to rule out cross-contamination from the control. Giardia detection rates were higher and TTP was lower ( P < 0.05) for 0.1% Alconox (19/20, 8.85 ± 0.3 min) compared with 1 M glycine (15/20, 14.53 ± 7.2 min). The LAMP assay detected 10 Giardia cysts spiked on leafy greens in 13–34 min in 14/40 samples tested. Robustness assessment showed that TTP was higher ( P < 0.0001) when spiked produce was stored for 7 days (13.09 ± 1.14 min) compared to fresh (9.72 ± 0.43 min). No unspiked samples were positive by LAMP, and the Tm for DNA of Giardia spiked on leafy greens was higher ( P < 0.0001, 87.43 ± 0.05 °C) than the APC (86.43 ± 0.12 °C). Within-assay repeatability co-efficient of variation (CV) for TTP was 5.4% and no cross-contamination occurred when spiked and un-spiked samples were processed in alternate order. The optimized sample processing procedure combined with the EF1α LAMP assay is a sensitive, specific, labour-saving, and rapid method for the detection of Giardia cysts in leafy greens.

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“…In a similar study, Jiang et al developed a facile, quick and responsive visual and POC detection approach for Cronobacter spp., Salmonella spp., and S. aureus in powdered infant formula (PIF) based on lateral flow dipstick (LFD) integrated with mLAMP [90]. Serious blood infections or meningitis and other inflammations of the brain and spine can result from Cronobacter bacteria [91].…”

Section: Gold Nanoparticlesmentioning

confidence: 99%

Progress in the Application of Nanoparticles and Graphene as Drug Carriers and on the Diagnosis of Brain Infections

et al. 2021

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The blood–brain barrier (BBB) is the protective sheath around the brain that protects the sensitive microenvironments of the brain. However, certain pathogens, viruses, and bacteria disrupt the endothelial barrier and cause infection and hence inflammation in meninges. Macromolecular therapeutics are unable to cross the tight junctions, thereby limiting their bioavailability in the brain. Recently, nanotechnology has brought a revolution in the field of drug delivery in brain infections. The nanostructures have high targeting accuracy and specificity to the receptors in the case of active targeting, which have made them the ideal cargoes to permeate across the BBB. In addition, nanomaterials with biomimetic functions have been introduced to efficiently cross the BBB to be engulfed by the pathogens. This review focuses on the nanotechnology-based drug delivery approaches for exploration in brain infections, including meningitis. Viruses, bacteria, fungi, or, rarely, protozoa or parasites may be the cause of brain infections. Moreover, inflammation of the meninges, called meningitis, is presently diagnosed using laboratory and imaging tests. Despite attempts to improve diagnostic instruments for brain infections and meningitis, due to its complicated and multidimensional nature and lack of successful diagnosis, meningitis appears almost untreatable. Potential for overcoming the difficulties and limitations related to conventional diagnostics has been shown by nanoparticles (NPs). Nanomedicine now offers new methods and perspectives to improve our knowledge of meningitis and can potentially give meningitis patients new hope. Here, we review traditional diagnosis tools and key nanoparticles (Au-NPs, graphene, carbon nanotubes (CNTs), QDs, etc.) for early diagnosis of brain infections and meningitis.

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Multiplex loop-mediated isothermal amplification-based lateral flow dipstick for simultaneous detection of 3 food-borne pathogens in powdered infant formula

Cited by 42 publications

References 36 publications

Optimizing Effective Parameters to Enhance the Sensitivity of Vertical Flow Assay for Detection of Escherichia coli

Optimizing Effective Parameters to Enhance the Sensitivity of Vertical Flow Assay for Detection of Escherichia coli

Optimization and validation of a loop-mediated isothermal amplification (LAMP) assay for detection of Giardia duodenalis in leafy greens

Progress in the Application of Nanoparticles and Graphene as Drug Carriers and on the Diagnosis of Brain Infections

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