Multiplex lateral flow assay development for snake venom detection in biological matrices

Abstract

Bothrops and Lachesis are two of Brazil’s medically most relevant snake genera, causing tens of thousands of bites annually. Fortunately, Brazil has good accessibility to high-quality antivenoms at the genus and inter-genus level, enabling the treatment of many of these envenomings. However, the optimal use of these treatments requires that the snake species responsible for the bite is determined. Currently, physicians use a syndromic approach to diagnose snakebite, which can be difficult for medical personnel with limited training in clinical snakebite management. In this work, we have developed a novel monoclonal antibody-based multiplex lateral flow assay for differentiating Bothrops and Lachesis venoms within 15 min. The test can be read by the naked eye or (semi)-quantitatively by a smartphone supported by a 3D-printed attachment for controlling lighting conditions. The LFA can detect Bothrops and Lachesis venoms in spiked plasma and urine matrices at concentrations spanning six orders of magnitude. The LFA has detection limits of 10–50 ng/mL in spiked plasma and urine, and 50–500 ng/mL in spiked sera, for B. atrox and L. muta venoms. This test could potentially support medical personnel in correctly diagnosing snakebite envenomings at the point-of-care in Brazil, which may help improve patient outcomes and save lives.

Molecular Memory Micromotors for Fast Snake Venom Toxin Dynamic Detection

“…Our approach presents several advantages over it since it can be applied to any type of venom (the traditional one is only available for specific Australian snakes) and does not suffer from cross-reactivity and low sensitivity . The LODs obtained with the MIPs MMs are like those obtained in LFA and paper-based optical assays for the determination of similar toxins. − Also, the LODs are within the range of a multiplexed fluorescence assay (1 mg/mL) . Remarkably, our moving-based assay has the lowest detection time reported (20 s) as compared with the 2 to 15 min required by the previously mentioned approaches.…”

“… 8 The LODs obtained with the MIPs MMs are like those obtained in LFA and paper-based optical assays for the determination of similar toxins. 13 − 18 Also, the LODs are within the range of a multiplexed fluorescence assay (1 mg/mL). 21 Remarkably, our moving-based assay has the lowest detection time reported (20 s) as compared with the 2 to 15 min required by the previously mentioned approaches.…”

“…Such tools are normally based on the use of specifically designed antibodies toward targeted venoms from different species, facilitating optical detection with portable smartphones in just 15 min. LFAs also provided a limit of detection that met the clinically relevant range postsnakebite (from 0.1 to 1000 ng/mL). − It should be mentioned here, yet, the performance can be affected by the matrix influence when applied to blood or serum samples . Paper-based optical assays have also been developed, using antibodies in connection with the enzyme horseradish peroxidase and the colorimetric substrate tetramethylbenzidine for β-bungarotoxin detection in blood, plasma, and urine.…”

Inoculum Size and False-Positive Detection of NDM- and OXA-48-Type Carbapenemases Using Two Multiplex Lateral Flow Assays

“…It is important to note that multiple species from three bacterial genera were involved. In LFAs, it is well known that false-positive results can be caused by the inadequate optimization of reaction buffers, nonspecific protein–conjugate interactions and cross-reactivity [ 27 , 28 ]. Our findings corroborate previous studies showing that the false-positive results mostly involved the NDM band for the CARBA5-LFA and both the NDM and OXA-48-like bands for the KNIVO-LFA, with the intensities of the false-positive bands being fainter than the control bands ( Table 3 ).…”

Retrospective Evaluation of Clinical and Clinicopathologic Findings, Case Management, and Outcome for Dogs and Cats Exposed to Micrurus fulvius (Eastern Coral Snake): 92 Cases (2021–2022)

“…Previous studies have called for point-of-care diagnostic tests specific to Micrurus fulvius venom. An ELISA test for coral snake venom has been previously described in people, and there is ongoing research on a multiplex lateral flow assay to differentiate different venomous snakes in Brazil [ 11 , 29 , 30 ]. This would allow for significant savings in resources (antivenom, cage space for patients under monitoring for development of clinical signs, nursing care, etc.)…”