Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark; BioPorto Diagnostics A/S, Hellerup, Denmark; VenomAid Diagnostics ApS, Kongens Lyngby, Denmark.
VenomAid Diagnostics ApS, Kongens Lyngby, Denmark.
Anal Chim Acta. 2023 Sep 1;1272:341306. doi: 10.1016/j.aca.2023.341306. Epub 2023 May 1.
Brazil is home to a multitude of venomous snakes; perhaps the most medically relevant of which belong to the Bothrops genus. Bothrops spp. are responsible for roughly 70% of all snakebites in Brazil, and envenomings caused by their bites can be treated with three types of antivenom: bothropic antivenom, bothro-lachetic antivenom, and bothro-crotalic antivenom. The choice to administer antivenom depends on the severity of the envenoming, while the choice of antivenom depends on availability and on how certain the treating physician is that the patient was bitten by a bothropic snake. The diagnosis of a bothropic envenoming can be made based on expert identification of the dead snake or a photo thereof or based on a syndromic approach wherein the clinician examines the patient for characteristic manifestations of envenoming. This approach can be very effective but requires staff that has been trained in clinical snakebite management, which, unfortunately, far from all relevant staff has.
In this article, we describe a prototype of the first lateral flow assay (LFA) capable of detecting venoms from Brazilian Bothrops spp. The monoclonal antibodies for the assay were generated using hybridoma technology and screened in sandwich enzyme-linked immunosorbent assays (ELISAs) to identify Bothrops spp.-specific antibody sandwich pairs. The prototype LFA is able to detect venom from several Bothrops spp. The LFA has a limit of detection (LoD) of 9.5 ng/mL in urine, when read with a commercial reader, and a visual LoD of approximately 25 ng/mL.
The work presented here serves as a proof of concept for a genus-specific venom detection kit that could support physicians in diagnosing Bothrops envenomings. Although further optimisation and testing is needed before the LFA can find clinical use, such a device could aid in decentralising antivenoms in the Brazilian Amazon and help ensure optimal snakebite management for even more victims of this highly neglected disease.
巴西是多种毒蛇的家园；也许最具医学相关性的毒蛇属于矛头蝮属。矛头蝮属蛇类约占巴西所有蛇伤的 70%，其咬伤引起的蛇伤可以用三种抗蛇毒血清治疗：矛头蝮抗蛇毒血清、矛头蝮溶血抗蛇毒血清和矛头蝮出血抗蛇毒血清。抗蛇毒血清的使用取决于中毒的严重程度，而抗蛇毒血清的选择取决于可用性以及治疗医生确定患者被矛头蝮蛇咬伤的确定性。根据专家对死蛇或其照片的识别，或根据临床医生检查患者是否存在蛇伤特征表现的综合征方法，可以诊断出矛头蝮蛇伤。这种方法非常有效，但需要经过临床蛇伤管理培训的工作人员，而不幸的是，并非所有相关工作人员都接受过这种培训。
在本文中，我们描述了第一个能够检测巴西矛头蝮属蛇类毒液的侧向流动分析（LFA）原型。该测定用的单克隆抗体是使用杂交瘤技术产生的，并在夹心酶联免疫吸附测定（ELISA）中进行了筛选，以鉴定矛头蝮属特异性抗体夹心对。原型 LFA 能够检测几种矛头蝮属蛇类的毒液。LFA 在使用商业阅读器读取时，尿液中的检测限（LoD）为 9.5ng/mL，视觉 LoD 约为 25ng/mL。
这里介绍的工作为一种属特异性毒液检测试剂盒提供了概念验证，该试剂盒可以帮助医生诊断矛头蝮蛇伤。虽然在侧向流动分析可以临床应用之前还需要进一步优化和测试，但这种设备可以帮助在巴西亚马逊地区分散抗蛇毒血清，并有助于确保更多这种被严重忽视疾病的受害者得到最佳的蛇伤管理。
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