Table 1.
Overview of snakebite diagnostics capable of differentiating snake venoms.
| Type | Subtype | Abs | Area | Snake(s) targeted | LoD | Assay duration | Tested on patient samples? | Sample matrix | Notes | Year | Ref. |
|---|---|---|---|---|---|---|---|---|---|---|---|
| Immuno-assay | Immuno-diffusion | Equine | Cobra spp. (possibly O. hannah) | 1:100,000,000 dilution | Patients: 1 (case study) | Tissue homogenate | 1957 | (24) | |||
| Immuno-diffusion | ? | Australia |
A. antarcticus, N. scutatus, P. porphyriacus, P. textilis |
? | >3 hours | ? | Serum exudate (guineapig) | Only abstract available | 1967 | (25) | |
| Agglutination test | Caprine & equine | California | C. v. helleri | >2 hours | Patients: 16 | Serum | 1968 | (26) | |||
| Immuno-diffusion | Leporine | Africa |
B. arietans, C. maculatus, E. carinatus, N. haje, N. melanoleuca, N. nigricollis |
48 hours (immuno-difussion) & >1 hour (immuno-electrophoresis) | Patients: 101 | Wound aspirates, blister fluids, sera, urine | Sensitivity: 39.6% (40/101) | 1974 | (27) | ||
| RIA | Australia |
N. scutatus, P. textilis |
<10 ng | >24 hours | Patients: 2 (also tested on rabbit serum) | Serum, sample buffer | 1974 | (28) | |||
| RIA | Australia |
N. scutatus, P. textilis |
>24 hours | Patients: 3 | Tissue samples, fluids | 1975 | (29) | ||||
| ELISA | Leporine & equine IgGs | Multiple |
B. arietans, C. maculatus, E. carinatus |
1-5 ng/mL | O/N incubation | No – Tested in rodents | Serum (human and rat) | Cross-reactivity to the following species tested: A. rhodostoma, B. gabonica, C. adamanteus, E. schistosa, N. haje, N. naja, N. nigricollis, O. scutellatus, V. berus | 1977 | (30) | |
| RIA | Leporine | Australia | N. scutatus | O/N incubation | Patients: 3 | Urine, serum, clothing, tissue samples | 1977 | (31, 32) | |||
| RIA | Leporine IgG | Australia |
A. antarcticus, A. superba, N. scutatus, O. scutellatus, P. australis, P. porphyriacus, P. textilis |
0.1-0.4 ng/mL | >20 hours | Unpublished | Tissue samples | 1978 | (33) | ||
| ELISA | Leporine IgG | ? | 0.5-2 ng/mL | 30-90 minutes | ? | ? | Only abstract available | 1980 | (34) | ||
| Enzyme immunoassay | Leporine IgG | Australia |
Acantophis spp., Notechis spp., Oxyuranus spp., Pseudoechis spp., Pseudonaja spp. |
5-15 ng/mL | 20-40 minutes | Patients: 43 | Whole blood, urine, wound swab | 1982 | (35) | ||
| ELISA | ? | ? | ? | 1 ng/mL | >3 hours | No - Tested in rabbits | Blood, urine, exudate | Only abstract available | 1983 | (36) | |
| ELISA | Leporine IgG | ? |
A. rhodostoma, N. naja |
7.8-15.6 ng/mL | 35-45 minutes | Yes | Serum | Only abstract available | 1983 | (37) | |
| ELISA | Leporine IgG | USA |
A. contortrix, C. atrox, C. scutulatus |
0.1-.01 µg/mL | O/N incubation | No - Tested in animals | Sero-sanguineous fluid, blood, urine, peritoneal fluid, pleural fluid, lung, kidney | Cross-reactions with venoms of other snakes were extensive at higher concentrations | 1984 | (38) | |
| ELISA | Equine | Myanmar | D. russelii | 10 ng/mL | O/N incubation | Yes | Serum | No cross-reactivity found to B. fasciatus, N. naja, O. hannah, T. gramineus | 1984 | (39) | |
| ELISA | Leporine | Thailand | N. kaouthia | 2 ng/well | Yes | Serum | 1986 | (40) | |||
| Enzyme-linked coagulation assay | Murine monoclonal IgG | D. russelii | 2-10 pg/mL | No | 1987 | (41) | |||||
| ELISA | Nigeria |
B. arietans, C. maculatus, E. carinatus |
O/N incubation | Patients: 31 | Blood, serum, urine, sputum, bite site aspirates | 1987 | (42) | ||||
| ELISA | Philippines | N. n. philippinensis | O/N incubation | Patients: 1 (postmortem) | Blood | 1987 | (43) | ||||
| ELISA | Leporine IgG | Thailand |
C. rhodostoma, D. russelii, N. kaouthia, T. albolabris |
10-20 ng/mL | O/N incubation | Patients: 251 | Serum | 1987 | (44) | ||
| RIA | Murine monoclonal Abs | Thailand | D. russelii | 4-20 ng/mL depending on matrix | O/N incubation | Patients: 4 | Serum, urine | Known to cross-react with cobra venom. | 1987 | (45) | |
| ELISA | Equine | Europe | V. ammodytes | <1 ng/mL | <20 minutes | No - Tested in rabbits | Blood | Specificity mentioned as being a problem. | 1988 | (46) | |
| ELISA | ? | Asia | A. b. blomhoffii | 5.4 ng/well | ? | No - Tested in mice | Serum | Only abstract available. No cross-reactivity to R. t. tigrinus venom. | 1988 | (47) | |
| ELISA | Leporine IgG | Brazil | B. jararaca | 14.6 ng/mL | O/N incubation | No – Tested in mice | Serum | Tested for cross-reactivity to Bothrops spp., Crotalus spp., Lachesis spp., and Tityus serrulatus venom. | 1990 | (48) | |
| ELISA | ? | ? | A. b. blomhoffii | ? | ? | No - Tested in rats & rabbits | Serum | Only abstract available | 1990 | (49) | |
| Agglutination test | Leporine IgG | Thailand |
B. fasciatus, C. rhodostoma, D. russelii, N. kaouthia, N. n. siamensis, O. hannah, T. albolabris |
0.16-1.2 µg/mL | 40 minutes | Serum samples: 59 Wound swabs: 26 |
Serum, wound swabs | Sensitivity of 52.5%. Tested for hook effect and interference from sample matrices. | 1991 | (50) | |
| ELISA | Equine F(ab’)2 | Brazil | C. d. terrificus | 1-3 pg/mL | O/N incubation | No – Tested in mice | Serum (mice), sample buffer | 1991 | (51) | ||
| ELISA | Leporine IgG | Myanmar | D. russelii | 10 ng/mL | O/N incubation | Patients: 311 Controls: 118 |
Serum | Specificity 88% (14 false positives from 118 negatives). Tested for cross-reactivity to B. fasciatus, N. kaouthia, O. hannah, T. erythrurus. | 1991 | (52) | |
| ELISA | Equine F(ab’)2 | Europe | Vipera spp. | 2-7 ng/mL (depending on sample matrix) | >4.5 hours | Yes | Blood, serum, urine | Tested for cross-reactivity to B. jararaca and C. d. terrificus. | 1992 | (53–56) | |
| ELISA | Leporine IgG | Australia |
A. antarcticus, N. scutatus, O. scutellatus, P. australis, P. textilis. |
2.5 ng/mL | O/N incubation | Unpublished | Sample buffer | 1992 | (57) | ||
| Leporine IgG | Southern Thailand | C. rhodostoma | 5 ng/mL | 50 minutes | No | Sample buffer | Cross-reactivity to 26 venoms tested. | 1992 | (58) | ||
| Agglutination test | Leporine IgG | Thailand |
B. fasciatus, C. rhodostoma, D. russellli, N. kaouthia, O. hannah, T. albolabris |
2-635 ng/mL | 60-120 minutes | Serum samples: 59 Wound swabs: 26 |
Serum, wound swab | Sensitivity of 81.3% for serum samples and 61.5% for wound swabs. Cross reactivity at concentrations at least 62 times higher. | 1993 | (59) | |
| ELISA | Leporine IgG | Brazil |
B. alternatus, B. atrox, B. jararaca, B.jararacussu, B. moojeni, B. neuwedi, C. d. terrificus, C. d. collineatus, L. muta |
<0.01-0.1 µg/mL | O/N incubation | No | Sample buffer, serum (non-envenomed humans) | 1993 | (60) | ||
| ELISA | Leporine IgG | Brazil |
B. atrox, L. m. muta |
20 ng/mL | 2 hours | Yes | Serum | Also tested in mice. | 1993 | (61) | |
| Fluorogenic ELISA | ? | D. russelii | 0.1 pg/mL | ? | ? | ? | Only abstract available. Shown to cross-react with several other venoms. | 1993 | (62) | ||
| ELISA | Papua New Guinea | P. papuanus | O/N incubation | Patients: 9 | Serum, urine, wound aspirates | 1994 | (63) | ||||
| ELISA | Leporine IgG | Tunisia | E. pyramidum | <10 ng/mL | O/N | Yes | Serum | 1994 | (64) | ||
| ELISA | North America | Agkistrodon spp. | 2 ng/mL | >5 hours | No – Tested in rabbits | Serum | 1994 | (65) | |||
| ELISA | Myanmar | O. hannah | <20 ng/mL | Patients: 2 (case studies) | Serum | 1995 | (66) | ||||
| ELISA & RIA | Ovine Fab | Europe | Vipera spp. | 0.8 ng/mL (ELISA) & 2 ng/mL (RIA) | >3 hours (ELISA) & O/N incubation (RIA) | Yes | Plasma, urine | 1996 | (67) | ||
| ELISA | Leporine F(ab’)2 | India |
B. caerulus, D. russelii, E. carinatus, N. naja |
1 ng/mL | 30 min | Patients: 27 | Blood, serum, urine, wound swab | Only abstract available | 1996 | (68) | |
| ELISA | Equine F(ab’)2 | Martinique | B. lanceolatus | 3 hours | Patients: 40 Controls: 120 |
Serum | Sensitivity 46%, specificity 88%. | 1997 | (69) | ||
| ELISA & agglutination assay | Equine | Central America | Micrurus spp. | 0.3 mg/mL (agglutination assay) & 4 ng/mL (ELISA) | >5 minutes (agglutination test) & O/N incubation (ELISA) | No – Tested in rabbits and mice | Serum, plasma | 1997 | (70) | ||
| ELISA | Caprine & leporine IgG | India |
Bungarus spp., Daboia spp., Echis spp., Naja spp. |
0.1 ng/mL | >5 hours | Yes (postmortem only) | Tissue samples | 1999 | (71) | ||
| ELISA | ? | Taiwan | Cobra spp. | 0.5 ng/mL | 6 hours | ? | Calf serum and human urine | Only abstract available | 2002 | (72) | |
| Optical immunoassay | Leporine IgG | Asia | B. multicinctus | 2.5-10 ng/mL | 25 minutes | No – Tested in mice | Blood, tissue samples | Cross-reactivity to 11 venoms and toxins tested. | 2002 | (73) | |
| Agglutination test | Equine | Venezuela |
Bothrops spp., Crotalus spp. |
167 µg/mL | 10 minutes | No | Sample buffer | LoD unit uncertain. | 2004 | (74) | |
| Optical immunoassay | Leporine IgG | Vietnam |
C. rhodostoma, N. kaouthia, O. hannah, T. albolabris |
0.2-0.8 ng/mL depending on the venom and sample matrix | 40 minutes | Patients: 83 Samples: 125 |
Serum, urine, wound exudate | 2004 | (75) | ||
| ELISA | Avian IgY & leporine IgG | India | N. naja | 0.1-300 ng | O/N incubation | Patients: 12 (live) Patients: 7 (postmortem) |
Skin, blood, cerebrospinal fluid | 2006 | (76) | ||
| ELISA | Leporine IgG | India |
B. caeruleus, N. naja |
O/N incubation | Samples: 22 (postmortem) | Skin, blood | 2007 | (77) | |||
| Immuno-flourescence | N. kaouthia | 5–10 ng/mL | 3 hours | No | Sample buffer | 2008 | (78) | ||||
| ELISA | Leporine IgG | Australasia | Oxyuranus spp. | 0.15 ng/mL | O/N incubation | Patients: 17 | Serum | Also tested in rat serum and for cross-reactivity with Australian snake venoms. | 2010 | (79) | |
| ELISA | Leporine IgG | Colombia | L. acrochorda | 3.9 ng/mL | No | Sample buffer | Specificity 100%. | 2012 | (80) | ||
| ELISA | Leporine IgG | Egypt |
N. haje, N. nigricollis, W. aegyptia |
<10 ng/well | O/N incubation | No | Sample buffer | Avidities of 2.5-2.8 depending on the venom | 2013 | (81) | |
| LFA | Avian | Taiwan | N. atra | 5 ng/mL | 20 minutes | Patients: 88 (34 cobra and 54 non-cobra) | Serum | Sensitivity 83.3%, specificity 100%. | 2014 | (82) | |
| LFA | Equine & leporine IgG | India | Daboia spp., Naja spp. | 0.1 ng/mL | 10 minutes | No - Tested in mice | Plasma | 2016 | (83) | ||
| ELISA | Leporine IgG | India |
Bungarus spp., Daboia spp., Echis spp., Naja spp. |
1 ng/mL | 20-25 minutes | No - Tested in mice | Sample buffer | 2017 | (84) | ||
| ELISA & LFA | Equine | Taiwan | Neurotoxic vs hemorrhagic venom | LoD of 5-50 ng/mL (LFA) & LoQ of 0.39-0.78 ng/mL (ELISA) | 10-15 minutes | Patients: 21 | Serum | Sensitivity and specificity of 100% for neurotoxic venoms. Sensitivity of 36.4% for hemorrhagic venoms. | 2018 | (85) | |
| Impedimetric immunoassay | Equine | Brazil | Bothrops spp. | 0.27 ug/mL | >25 minutes? | No | Sample buffer | Tested for cross-reactivity to C. d. terrificus and M. leminiscatus. | 2018 | (86) | |
| ELISA | Leporine | Sri Lanka |
B. caeruleus, D. russelii, H. hypnale, N. naja |
0.19-1.56 ng/mL (depending on the venom) | >2 hours | Patients: 19 Controls: 20 |
Serum | Quantitative. Cross-reactivity between the venoms was tested. | 2020 | (87) | |
| LFA | Avian & equine | South-East Asia |
Daboia spp., Naja spp. |
10 ng/mL (in vitro) | 25-30 minutes | Samples: 5 | Serum | 2020 | (88) | ||
| LFA | Leporine & equine | Asia & Africa | Naja spp. | 5-10 ng/mL for Asian cobras and <500 ng/mL for African cobras | >20 minutes | No | Serum (fetal bovine) | Based on (82). | 2020 | (89) | |
| Molecular biology | PCR | N/A | Thailand | N. kaouthia | >2 hours | No - Tested on mice | Wound swabs (mice) | Also tested on venom from B. fasciatus, C. rhadostma, D. russelii, O. Hannah. | 2001 | (90) | |
| PCR | N/A | Thailand |
B. fasciatus, C. rhodostoma, D. siamensis, Hydrophiinae spp., Naja spp., O. Hannah, Trimeresurus spp. |
0.025 ng/mL | >67 minutes | No | Saliva (snake) | 2015 | (91) | ||
| PCR | N/A | Nepal |
Bungarus spp., Naja spp., O. hannah, O. moticola, Trimeresurus spp. |
O/N incubation | Patients: 565 | Wound swab | Specificity 100%. | 2016 | (92) | ||
| Misc. | Enzymatic activity assay | N/A | Sri Lanka & Australia |
B. caeruleus, D. russelii, H. hypnale, N. naja, P. porphyriacus |
Patients: 108 | Serum | 2014 | (93) | |||
| Infrared thermography | N/A | Brazil |
B. moojeni, C. d. terrificus, B. jararaca |
>15 minutes | Patients: 8 | 2017 | (94) | ||||
| Enzymatic activity assay & ELISA | N/A | Australia | Elapid spp. | 0.1-0.2 ng/mL (ELISA) | Patients: 115 Controls: 80 |
Serum | 2018 | (95) |
Many studies did not report on the duration of the diagnostic procedure. In such cases, assay duration was reported in this table as “> total incubation time”, e.g. “>3 hours” for an assay with an incubation time of 3 hours. For other unreported values, the corresponding fields were left empty. In some cases, only the abstracts of the studies were available to us, and in these cases, values not reported in the abstract have been marked “?”. Studies describing the detection of venom-specific antibodies in snakebite victims [e.g. (96–98)] or the detection of toxins or toxin activities for the purpose of venom characterization rather than diagnosis [e.g. (99–103)] were not included in this table. Abs, antibodies; ELISA, enzyme-linked immunosorbent assay; F(ab’)2, fragment antigen binding 2; IgG, immunoglobulin G; LFA, lateral flow assay; O/N, overnight; PCR, polymerase chain reaction; RIA, radioimmunoassay.