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...
Main Authors: | , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2022-01-01
|
Series: | Molecules |
Subjects: | |
Online Access: | https://www.mdpi.com/1420-3049/27/3/888 |
_version_ | 1797486103649845248 |
---|---|
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. |
first_indexed | 2024-03-09T23:28:27Z |
format | Article |
id | doaj.art-9823c537c5224d8699bb530440079f96 |
institution | Directory Open Access Journal |
issn | 1420-3049 |
language | English |
last_indexed | 2024-03-09T23:28:27Z |
publishDate | 2022-01-01 |
publisher | MDPI AG |
record_format | Article |
series | Molecules |
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 |
work_keys_str_mv | AT ernestzmanson developmentofaninhibitionenzymelinkedimmunosorbentassayelisaprototypefordetectingcytotoxicthreefingertoxins3ftxsinafricanspittingcobravenoms AT kyamacmutinda developmentofaninhibitionenzymelinkedimmunosorbentassayelisaprototypefordetectingcytotoxicthreefingertoxins3ftxsinafricanspittingcobravenoms AT josephkgikunju developmentofaninhibitionenzymelinkedimmunosorbentassayelisaprototypefordetectingcytotoxicthreefingertoxins3ftxsinafricanspittingcobravenoms AT aleksandrabocian developmentofaninhibitionenzymelinkedimmunosorbentassayelisaprototypefordetectingcytotoxicthreefingertoxins3ftxsinafricanspittingcobravenoms AT konradkhus developmentofaninhibitionenzymelinkedimmunosorbentassayelisaprototypefordetectingcytotoxicthreefingertoxins3ftxsinafricanspittingcobravenoms AT vladimirpetrilla developmentofaninhibitionenzymelinkedimmunosorbentassayelisaprototypefordetectingcytotoxicthreefingertoxins3ftxsinafricanspittingcobravenoms AT jaroslavlegath developmentofaninhibitionenzymelinkedimmunosorbentassayelisaprototypefordetectingcytotoxicthreefingertoxins3ftxsinafricanspittingcobravenoms AT jameshkimotho developmentofaninhibitionenzymelinkedimmunosorbentassayelisaprototypefordetectingcytotoxicthreefingertoxins3ftxsinafricanspittingcobravenoms |