Enzyme immunoassays in brown snake (Pseudonaja spp.) envenoming: Detecting venom, antivenom and venom-antivenom complexes

M O'LEARY, Geof Isbister, Jennifer Schneider, S BROWN, Bart Currie

    Research output: Contribution to journalArticle

    Abstract

    Although a commercial snake venom detection kit (SVDK) is available to distinguish between the five major snake groups in Australia, there is no assay for quantifying venom or antivenom concentrations in envenomed patients. Serum samples were obtained from patients with brown snake (Pseudonaja spp.) envenoming before and after the administration of antivenom and patients with suspected brown snake bites but no evidence of envenoming. Enzyme immunoassays (EIAs) were developed for free venom, free antivenom and the venom-antivenom complex. Standard samples measured in duplicate had a coefficient of variation of less than 10%. The EIA for venom was able to detect brown snake venom down to concentrations of 3 ng/mL. A high baseline absorbance was measured in some patients that did not change with the addition of excess antivenom to the samples. In these patients, the baseline absorbance was subtracted from all measurements to calculate the true venom concentration. The EIA for brown snake antivenom had a limit of detection of 20 ?g/mL, but 50 ?g/mL was used as a cut-off based on assays in patients who had not received antivenom. The EIA for venom-antivenom complexes was unable to detect these at the low venom concentrations that occurred in patients. Quantification of venom and antivenom will help to determine the dose of antivenom required to bind venom and to establish appropriate end points for antivenom treatment. � 2006 Elsevier Ltd. All rights reserved.
    Original languageEnglish
    Pages (from-to)4-11
    Number of pages8
    JournalToxicon
    Volume48
    Issue number1
    Publication statusPublished - 2006

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    Antivenins
    Snakes
    Venoms
    Immunoenzyme Techniques
    Enzymes
    Snake Venoms
    Assays
    Snake Bites
    Limit of Detection

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    O'LEARY, M ; Isbister, Geof ; Schneider, Jennifer ; BROWN, S ; Currie, Bart. / Enzyme immunoassays in brown snake (Pseudonaja spp.) envenoming : Detecting venom, antivenom and venom-antivenom complexes. In: Toxicon. 2006 ; Vol. 48, No. 1. pp. 4-11.
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    abstract = "Although a commercial snake venom detection kit (SVDK) is available to distinguish between the five major snake groups in Australia, there is no assay for quantifying venom or antivenom concentrations in envenomed patients. Serum samples were obtained from patients with brown snake (Pseudonaja spp.) envenoming before and after the administration of antivenom and patients with suspected brown snake bites but no evidence of envenoming. Enzyme immunoassays (EIAs) were developed for free venom, free antivenom and the venom-antivenom complex. Standard samples measured in duplicate had a coefficient of variation of less than 10{\%}. The EIA for venom was able to detect brown snake venom down to concentrations of 3 ng/mL. A high baseline absorbance was measured in some patients that did not change with the addition of excess antivenom to the samples. In these patients, the baseline absorbance was subtracted from all measurements to calculate the true venom concentration. The EIA for brown snake antivenom had a limit of detection of 20 ?g/mL, but 50 ?g/mL was used as a cut-off based on assays in patients who had not received antivenom. The EIA for venom-antivenom complexes was unable to detect these at the low venom concentrations that occurred in patients. Quantification of venom and antivenom will help to determine the dose of antivenom required to bind venom and to establish appropriate end points for antivenom treatment. � 2006 Elsevier Ltd. All rights reserved.",
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    Enzyme immunoassays in brown snake (Pseudonaja spp.) envenoming : Detecting venom, antivenom and venom-antivenom complexes. / O'LEARY, M; Isbister, Geof; Schneider, Jennifer; BROWN, S; Currie, Bart.

    In: Toxicon, Vol. 48, No. 1, 2006, p. 4-11.

    Research output: Contribution to journalArticle

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    T1 - Enzyme immunoassays in brown snake (Pseudonaja spp.) envenoming

    T2 - Detecting venom, antivenom and venom-antivenom complexes

    AU - O'LEARY, M

    AU - Isbister, Geof

    AU - Schneider, Jennifer

    AU - BROWN, S

    AU - Currie, Bart

    PY - 2006

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    N2 - Although a commercial snake venom detection kit (SVDK) is available to distinguish between the five major snake groups in Australia, there is no assay for quantifying venom or antivenom concentrations in envenomed patients. Serum samples were obtained from patients with brown snake (Pseudonaja spp.) envenoming before and after the administration of antivenom and patients with suspected brown snake bites but no evidence of envenoming. Enzyme immunoassays (EIAs) were developed for free venom, free antivenom and the venom-antivenom complex. Standard samples measured in duplicate had a coefficient of variation of less than 10%. The EIA for venom was able to detect brown snake venom down to concentrations of 3 ng/mL. A high baseline absorbance was measured in some patients that did not change with the addition of excess antivenom to the samples. In these patients, the baseline absorbance was subtracted from all measurements to calculate the true venom concentration. The EIA for brown snake antivenom had a limit of detection of 20 ?g/mL, but 50 ?g/mL was used as a cut-off based on assays in patients who had not received antivenom. The EIA for venom-antivenom complexes was unable to detect these at the low venom concentrations that occurred in patients. Quantification of venom and antivenom will help to determine the dose of antivenom required to bind venom and to establish appropriate end points for antivenom treatment. � 2006 Elsevier Ltd. All rights reserved.

    AB - Although a commercial snake venom detection kit (SVDK) is available to distinguish between the five major snake groups in Australia, there is no assay for quantifying venom or antivenom concentrations in envenomed patients. Serum samples were obtained from patients with brown snake (Pseudonaja spp.) envenoming before and after the administration of antivenom and patients with suspected brown snake bites but no evidence of envenoming. Enzyme immunoassays (EIAs) were developed for free venom, free antivenom and the venom-antivenom complex. Standard samples measured in duplicate had a coefficient of variation of less than 10%. The EIA for venom was able to detect brown snake venom down to concentrations of 3 ng/mL. A high baseline absorbance was measured in some patients that did not change with the addition of excess antivenom to the samples. In these patients, the baseline absorbance was subtracted from all measurements to calculate the true venom concentration. The EIA for brown snake antivenom had a limit of detection of 20 ?g/mL, but 50 ?g/mL was used as a cut-off based on assays in patients who had not received antivenom. The EIA for venom-antivenom complexes was unable to detect these at the low venom concentrations that occurred in patients. Quantification of venom and antivenom will help to determine the dose of antivenom required to bind venom and to establish appropriate end points for antivenom treatment. � 2006 Elsevier Ltd. All rights reserved.

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