Development of an Inhibition Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detecting Cytotoxic Three-Finger Toxins (3FTxs) in African Spitting Cobra Venoms

The administration of toxin-specific therapy in snake envenoming is predicated on improved diagnostic techniques capable of detecting specific venom toxins. Various serological tests have been used in detecting snakebite envenoming. Comparatively, enzyme-linked immunosorbent assay (ELISA) has been s...

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Main Authors: Ernest Z. Manson, Kyama C. Mutinda, Joseph K. Gikunju, Aleksandra Bocian, Konrad K. Hus, Vladimír Petrílla, Jaroslav Legáth, James H. Kimotho
Format: Article
Language:English
Published: MDPI AG 2022-01-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/27/3/888
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author Ernest Z. Manson
Kyama C. Mutinda
Joseph K. Gikunju
Aleksandra Bocian
Konrad K. Hus
Vladimír Petrílla
Jaroslav Legáth
James H. Kimotho
author_facet Ernest Z. Manson
Kyama C. Mutinda
Joseph K. Gikunju
Aleksandra Bocian
Konrad K. Hus
Vladimír Petrílla
Jaroslav Legáth
James H. Kimotho
author_sort Ernest Z. Manson
collection DOAJ
description The administration of toxin-specific therapy in snake envenoming is predicated on improved diagnostic techniques capable of detecting specific venom toxins. Various serological tests have been used in detecting snakebite envenoming. Comparatively, enzyme-linked immunosorbent assay (ELISA) has been shown to offer a wider practical application. We report an inhibition ELISA for detecting three-finger toxin (3FTx) proteins in venoms of African spitting cobras. The optimized assay detected 3FTxs in <i>N. ashei</i> (including other <i>Naja</i> sp.) venoms, spiked samples, and venom-challenged mice samples. In venoms of <i>Naja</i> sp., the assay showed inhibition, implying the detection of 3FTxs, but showed little or no inhibition in non-<i>Naja</i> sp. In mice-spiked samples, one-way ANOVA results showed that the observed inhibition was not statistically significant between spiked samples and negative control (<i>p</i>-value = 0.164). Similarly, the observed differences in inhibition between venom-challenged and negative control samples were not statistically significant (<i>p</i>-value = 0.9109). At an LOD of 0.01 µg/mL, the assay was able to confirm the presence of 3FTxs in the samples. Our results show a proof of concept for the use of an inhibition ELISA model as a tool for detecting 3FTxs in the venoms of African spitting cobra snakes.
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spelling doaj.art-9823c537c5224d8699bb530440079f962023-11-23T17:14:07ZengMDPI AGMolecules1420-30492022-01-0127388810.3390/molecules27030888Development of an Inhibition Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detecting Cytotoxic Three-Finger Toxins (3FTxs) in African Spitting Cobra VenomsErnest Z. Manson0Kyama C. Mutinda1Joseph K. Gikunju2Aleksandra Bocian3Konrad K. Hus4Vladimír Petrílla5Jaroslav Legáth6James H. Kimotho7Institute for Basic Sciences, Technology & Innovation, Pan African University, Nairobi 00100, KenyaDepartment of Medical Laboratory Science, College of Health Sciences, Jomo Kenyatta University of Agriculture & Technology, Nairobi 00100, KenyaDepartment of Medical Laboratory Science, College of Health Sciences, Jomo Kenyatta University of Agriculture & Technology, Nairobi 00100, KenyaDepartment of Biotechnology and Bioinformatics, Faculty of Chemistry, Rzeszow University of Technology, 35-959 Rzeszow, PolandDepartment of Biotechnology and Bioinformatics, Faculty of Chemistry, Rzeszow University of Technology, 35-959 Rzeszow, PolandDepartment of Biology and Physiology, University of Veterinary Medicine and Pharmacy, 041 81 Košice, SlovakiaDepartment of Biotechnology and Bioinformatics, Faculty of Chemistry, Rzeszow University of Technology, 35-959 Rzeszow, PolandKenya Medical Research Institute, Nairobi 00100, KenyaThe administration of toxin-specific therapy in snake envenoming is predicated on improved diagnostic techniques capable of detecting specific venom toxins. Various serological tests have been used in detecting snakebite envenoming. Comparatively, enzyme-linked immunosorbent assay (ELISA) has been shown to offer a wider practical application. We report an inhibition ELISA for detecting three-finger toxin (3FTx) proteins in venoms of African spitting cobras. The optimized assay detected 3FTxs in <i>N. ashei</i> (including other <i>Naja</i> sp.) venoms, spiked samples, and venom-challenged mice samples. In venoms of <i>Naja</i> sp., the assay showed inhibition, implying the detection of 3FTxs, but showed little or no inhibition in non-<i>Naja</i> sp. In mice-spiked samples, one-way ANOVA results showed that the observed inhibition was not statistically significant between spiked samples and negative control (<i>p</i>-value = 0.164). Similarly, the observed differences in inhibition between venom-challenged and negative control samples were not statistically significant (<i>p</i>-value = 0.9109). At an LOD of 0.01 µg/mL, the assay was able to confirm the presence of 3FTxs in the samples. Our results show a proof of concept for the use of an inhibition ELISA model as a tool for detecting 3FTxs in the venoms of African spitting cobra snakes.https://www.mdpi.com/1420-3049/27/3/888inhibition ELISAthree-finger toxinscytotoxicsnakebite envenomingdiagnostics
spellingShingle Ernest Z. Manson
Kyama C. Mutinda
Joseph K. Gikunju
Aleksandra Bocian
Konrad K. Hus
Vladimír Petrílla
Jaroslav Legáth
James H. Kimotho
Development of an Inhibition Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detecting Cytotoxic Three-Finger Toxins (3FTxs) in African Spitting Cobra Venoms
Molecules
inhibition ELISA
three-finger toxins
cytotoxic
snakebite envenoming
diagnostics
title Development of an Inhibition Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detecting Cytotoxic Three-Finger Toxins (3FTxs) in African Spitting Cobra Venoms
title_full Development of an Inhibition Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detecting Cytotoxic Three-Finger Toxins (3FTxs) in African Spitting Cobra Venoms
title_fullStr Development of an Inhibition Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detecting Cytotoxic Three-Finger Toxins (3FTxs) in African Spitting Cobra Venoms
title_full_unstemmed Development of an Inhibition Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detecting Cytotoxic Three-Finger Toxins (3FTxs) in African Spitting Cobra Venoms
title_short Development of an Inhibition Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detecting Cytotoxic Three-Finger Toxins (3FTxs) in African Spitting Cobra Venoms
title_sort development of an inhibition enzyme linked immunosorbent assay elisa prototype for detecting cytotoxic three finger toxins 3ftxs in african spitting cobra venoms
topic inhibition ELISA
three-finger toxins
cytotoxic
snakebite envenoming
diagnostics
url https://www.mdpi.com/1420-3049/27/3/888
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