| Summary: | 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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