Abstract
As said by former United Nations Secretary-General Kofi Annan, “Snakebite is the most important tropical disease you’ve never heard of.” Listed as a priority neglected tropical disease by the World Health Organization, snakebite envenoming (SBE) kills in excess of 125,000 people per year. However, due to the complexity and overlap of snake venom compositions, few reliable venom diagnostic methods for genus-/species-specific identification, which is crucial for successful SBE therapy, are available. Here, we develop a strategy to select and prepare genus-specific snake venom antibodies, which allows rapid and efficient clinical diagnosis of snakebite. Multi-omics approaches are used to choose candidate antigens from snake venoms and identify genus-specific antigenic epitope peptide fragments (GSAEPs) with ideal immunogenicity, specificity, and spatial accessibility. Double-antibody sandwich ELISA kit was established by matching a polyclonal antibody against a natural antigen and a monoclonal antibody that was prepared by natural protein as antigen and can specifically target the GSAEPs. The kit shows the ability to accurately identify venoms from similar genera of Trimeresurus and Protobothrops with a detection limit of 6.25 ng/ml on the snake venoms and a little cross-reaction, thus proving high feasibility and applicability.
Highlights
There are about 3,596 species of snakes worldwide and 768 of them are venomous
We propose a strategy for the efficient preparation of genus-specific diagnostic antibodies for snakebites, as follows: 1) data mining and multi-omics analysis of snake venom glands were performed to obtain information on proteins in different snake venoms and select potential antigens, which are highly abundant proteins containing ideal antigenic epitopes in various genera; 2) antigenic epitope peptide fragments, whose immunogenicity, specificity, and spatial accessibility are validated and verified, and natural venomous proteins containing the antigenic epitopes are used for the preparation of antibodies; and 3) the antibodies were used for the establishment of double-antibody sandwich enzymelinked immunosorbent assay (ELISA) to identify and verify venoms from different snakes (Figure 1)
The amino acid sequences of PLA2 and SVMP were analyzed by a BLAST search of the NCBI database to identify genus-specific peptide antigens (Supplementary Figures 1, 2)
Summary
There are about 3,596 species of snakes worldwide and 768 of them are venomous. Elapidae (elapids, consisting of nearly 300 species and 60 genera) and Viperidae (vipers, 329 species and 27 genera) are two strictly venomous families. Some species from the families of Colubridae (colubrids) and Lamprophiidae (lamprophiids) are venomous, most of the members. Most of the lethal cases are almost exclusively resulting from snakebite envenoming (SBE) of Viperidae and Elapidae [1, 2]. There are about 5.4 million snakebites annually, which results in 1.8 to 2.7 million SBE cases and at least 125,000 deaths [3]. Due to many incidences occurring in rural areas with poor health and transportation facilities, it is difficult to accurately estimate the global rates of snakebite and associated mortality, likely underreporting worldwide estimates
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