Collocation of avian and mammal antibodies to develop a rapid and sensitive diagnostic tool for Russell's Vipers Snakebite

Abstract

Russell's vipers (RVs) envenoming is an important public health issue in SouthEast Asia. Disseminated intravascular coagulopathy, systemic bleeding, hemolysis, and acute renal injury are obvious problems that develop in most cases, and neuromuscular junction blocks are an additional problem caused by western RV snakebite. The complex presentations usually are an obstacle to early diagnosis and antivenom administration. Here, we tried to produce highly specific antibodies in goose yolks for use in a paper-based microfluidic diagnostic kit, immunochromatographic test of viper (ICT-Viper), to distinguish RVs from other vipers and even cobra snakebite in Asia. We used indirect ELISA to monitor specific goose IgY production and western blotting to illustrate the interaction of avian or mammal antibody with venom proteins. The ICT-Viper was tested not only in prepared samples but also in stored patient serum to demonstrate its preliminary efficacy. The results revealed that specific anti-Daboia russelii IgY could be raised in goose eggs effectively without inducing adverse effects. When it was collocated with horse anti-Daboia siamensis antibody, which broadly reacted with most of the venom proteins of both types of Russell's viper, the false cross-reactivity was reduced, and the test showed good performance. The limit of detection was reduced to 10 ng/ml in vitro, and the test showed good detection ability in clinical snake envenoming case samples. The ICT-Viper performed well and could be combined with a cobra venom detection kit (ICT-Cobra) to create a multiple detection strip (ICT-VC), which broadens its applications while maintaining its detection ability for snake envenomation identification. Nonetheless, the use of the ICT-Viper in the SouthEast Asia region is pending additional laboratory and field investigations and regional collaboration. We believe that the development of this practical diagnostic tool marks the beginning of positive efforts to face the global snakebite issue.

Local Cytotoxic Effects in Cobra Envenoming: A Pilot Study

“…The horse anti-N. atra F(ab') 2 was obtained from the Centers for Disease Control (CDC), Taiwan. The horse and rabbit polyclonal antibody was affinity-purified using a Taiwan cobra venom-immobilized column prepared as described in our previous study [51]. Goat anti-horse IgG (H+L)-HRP and goat anti-mouse IgG (H+L)-HRP, the secondary antibodies, were purchased from Jackson ImmunoResearch Inc. (West Grove, PA, USA).…”

“…The sandwich ELISA was modified and applied to measure the concentration of whole venom (N. atra) as well as the levels of CTX A3 and sNTX in serum or wound discharge following the protocol that we previously developed and reported [15,20,51]. One day before analysis, the capture antibody (rabbit anti-N. atra IgG, 0.4 µg/mL) was coated onto a 96-well polystyrene microplate with a coating buffer (50 mM carbonate/bicarbonate buffer, pH 9.6) at 4 • C overnight.…”

Clinical management of snakebite envenoming: Future perspectives

“…Snakebite diagnostics reported in the literature include assays based on infrared thermography ( de Madeiros et al, 2017 ; Sabitha et al, 2021 ), measurement of enzymatic activity of toxins ( Maduwage et al, 2014 ), molecular biology methods ( Suntrarachun et al, 2001 ; Supikamolseni et al, 2015 ), and immunoassays (e.g., Faria et al, 2018 ; Lin et al, 2020 ; Hung et al, 2014 ; Liu et al, 2018 ) to mention only a few), of which enzyme-linked-immunosorbent assays (ELISAs) are the most frequently described. The pros and cons of these methods make them differentially suited for different uses.…”

Cerebral Complications of Snakebite Envenoming: Case Studies

“…Daboia russelii , also called Russell’s viper, is one of the most common vipers found in India [ 49 ] and is the greatest contributor to accidental snakebites [ 50 ]. To shorten the time needed to treat an unidentified snakebite, a group in Taiwan utilized avian and mammal antibodies to develop a rapid and sensitive diagnostic tool for a Russell’s viper’s snakebite [ 51 ]. Although D. russelii belongs to the family of Viperidae , several neurotoxins are identified in the viper’s venom [ 52 ] which also cause neural disorders such as ptosis and ophthalmoplegia [ 12 , 38 ].…”