Noncompetitive Chromogenic Lateral-Flow Immunoassay for Simultaneous Detection of Microcystins and Nodularin

Akter, Sultana; Kustila, Teemu; Leivo, Janne; Muralitharan, Gangatharan; Vehniäinen, Markus; Lamminmäki, Urpo

doi:10.3390/bios9020079

Cited by 18 publications

(4 citation statements)

References 24 publications

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“…There are other antibody-based platforms that can detect the dengue NS1 protein, some studies even demonstrate the use of enzymatic enhancement to increase the sensitivity by up to 10-fold compared to those of gold nanoparticles. However, not all of them reach the sensitivity shown in the paper; they require more than one step to operate, usually substrate addition; ,− they need sophisticated machinery; or they are able to give qualitative but not quantitative , results.…”

Section: Resultsmentioning

confidence: 97%

Capture-Layer Lateral Flow Immunoassay: A New Platform Validated in the Detection and Quantification of Dengue NS1

2020

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The lateral flow immunoassay (LFIA) is the most successful point-of-care testing (POCT) method to date. In the case of clinical biomarkers that require quantification, it remains a challenge to quantitate those biomarkers using the lateral flow immunoassay remains a challenge due to the cost of the reader and possibly the type of marker used. In the present work, a new concept of a platform LFIA device configuration is proposed in which different, aligned membrane components, some already existing in the classical lateral flow immunoassay, and the others created with special new functions in the present device. As the sample containing the target analyte passes through the aforementioned membranes, the target analyte will initially interact with a target-specific antibody-conjugated to horseradish peroxidase (HRP). Thereafter, the newly formed immunocomplex will diffuse through a proprietary capture membrane (that ensures that the nontarget-bound antibodies do not continue further and thus remain “captured” to that specific area). This is done by having the target molecules (or components thereof) immobilized onto the said capture layer. The target-bound immunocomplexes will then be allowed by the system configuration to continue further to the last layer, where the signal will be generated and quantified. Thus, in the absence of the target analyte in the sample, the free antibodies will be filtered at the capture layer by preimmobilized analyte molecules, thus preventing a false positive signal to occur. We validated the concept in the detection of dengue NS1 protein in view of making a triage test. The sample containing NS1 will first meet HRP-conjugated NS1-specific antibodies and become attached, thus producing an NS1-specific antibody–HRP immunocomplex. The sample then flows through the blocking layer, where the immunocomplex is unchallenged and thus allowed to reach the last “absorbent” pad, incorporating the substrate for the HRP marker. In the case of a positive test, a signal is generated, that is proportional to the amount of immunocomplexes (and therefore the NS1 concentration), and then analyzed and measured at the absorbent pad. Any unbound anti-NS1 antibody will be stopped at the blocking matrix by preimmobilized NS1, so there will be no false positive. As this study is the initial study of a novel configuration, much of the work comprised of optimization steps, such as determining the required NS1 membrane-immobilization concentration and the required target-specific capture antibody concentration. Our immunoassay was tested with spiked buffer and serum samples to mimic the clinical conditions, with a range of NS1 concentrations, and was found, at this time, to be fivefold more sensitive than a gold standard enzyme-linked immunosorbent assay (ELISA) test (5 ng mL–1) performed in our laboratory. This method shows another form of LFIA that has the potential to be quantitative (at least semiquantitative), albeit not solving the reader cost; however, unlike the regular LFIA, we do not use nanobeads but...

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Section: Resultsmentioning

confidence: 97%

Capture-Layer Lateral Flow Immunoassay: A New Platform Validated in the Detection and Quantification of Dengue NS1

2020

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“…The POC devices are replacing culture-based and other cumbersome commercial methods, which is attributable to widened accessibility to detection, rapid analysis time, and cost effectiveness [42]. Various POCs are majorly developed on microfluidic [43,44,45] or lateral flow systems [46,47,48] using glass; paper; silk fibroin; flexible graphite; or polymer substrates, such as polydimethylsiloxane (PDMS), poly(3,4-ethylene dioxythiophene):poly(styrene sulfonate) (PEDOT:PSS)), poly(methyl methacrylate) (PMMA), and polyethylene terephthalate (PET), etc. Further, the detection approaches are dependent upon fluorescence [49], colorimetry [50], and electrochemistry [51,52,53] to achieve rapid and easy to interpret signals for water safety monitoring.…”

Section: Point-of-care (Poc) Devices For Biological Contaminantsmentioning

confidence: 99%

Point-of-Care Strategies for Detection of Waterborne Pathogens

Kumar

Nehra

Mehta

et al. 2019

Sensors

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Waterborne diseases that originated due to pathogen microorganisms are emerging as a serious global health concern. Therefore, rapid, accurate, and specific detection of these microorganisms (i.e., bacteria, viruses, protozoa, and parasitic pathogens) in water resources has become a requirement of water quality assessment. Significant research has been conducted to develop rapid, efficient, scalable, and affordable sensing techniques to detect biological contaminants. State-of-the-art technology-assisted smart sensors have improved features (high sensitivity and very low detection limit) and can perform in a real-time manner. However, there is still a need to promote this area of research, keeping global aspects and demand in mind. Keeping this view, this article was designed carefully and critically to explore sensing technologies developed for the detection of biological contaminants. Advancements using paper-based assays, microfluidic platforms, and lateral flow devices are discussed in this report. The emerging recent trends, mainly point-of-care (POC) technologies, of water safety analysis are also discussed here, along with challenges and future prospective applications of these smart sensing technologies for water health diagnostics.

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“…Ouyang et al [ 49 ] reported a NOD-R detection limit as low as 167 pM using a newly developed DNA-based aptasensor with high selectivity and good reproducibility and stability. A more recent study [ 50 ] proposes a simple chromogenic lateral-flow immunoassay (LFIA) approach for the simultaneous detection of MC and NOD-R concentrations lower than 4 μg/L. One of the advantages of this reported detection method is the possibility to visually confirm the result without using additional measuring devices.…”

Section: Introductionmentioning

confidence: 99%

“… Main algal toxins correlated with blooming, and the most relevant techniques used for their detection as reported between 2017 and 2022 (source—Web of Science Collection). A table summarizing the lowest LODs obtained by using label-based approaches or elaborated platforms designed for trace-level detection [ 30 , 31 , 32 , 36 , 48 , 49 , 50 , 54 , 57 , 63 , 64 , 65 , 67 ]. …”

Section: Introductionmentioning

confidence: 99%

Detection and Characterization of Nodularin by Using Label-Free Surface-Enhanced Spectroscopic Techniques

Brezeștean

Gherman

Colniță

et al. 2022

IJMS

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Nodularin (NOD) is a potent toxin produced by Nodularia spumigena cyanobacteria. Usually, NOD co-exists with other microcystins in environmental waters, a class of cyanotoxins secreted by certain cyanobacteria species, which makes identification difficult in the case of mixed toxins. Herein we report a complete theoretical DFT-vibrational Raman characterization of NOD along with the experimental drop-coating deposition Raman (DCDR) technique. In addition, we used the vibrational characterization to probe SERS analysis of NOD using colloidal silver nanoparticles (AgNPs), commercial nanopatterned substrates with periodic inverted pyramids (KlariteTM substrate), hydrophobic Tienta® SpecTrimTM slides, and in-house fabricated periodic nanotrenches by nanoimprint lithography (NIL). The 532 nm excitation source provided more well-defined bands even at LOD levels, as well as the best performance in terms of SERS intensity. This was reflected by the results obtained with the KlariteTM substrate and the silver-based colloidal system, which were the most promising detection approaches, providing the lowest limits of detection. A detection limit of 8.4 × 10−8 M was achieved for NOD in solution by using AgNPs. Theoretical computation of the complex vibrational modes of NOD was used for the first time to unambiguously assign all the specific vibrational Raman bands.

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Noncompetitive Chromogenic Lateral-Flow Immunoassay for Simultaneous Detection of Microcystins and Nodularin

Cited by 18 publications

References 24 publications

Capture-Layer Lateral Flow Immunoassay: A New Platform Validated in the Detection and Quantification of Dengue NS1

Capture-Layer Lateral Flow Immunoassay: A New Platform Validated in the Detection and Quantification of Dengue NS1

Point-of-Care Strategies for Detection of Waterborne Pathogens

Detection and Characterization of Nodularin by Using Label-Free Surface-Enhanced Spectroscopic Techniques

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