A new assay for the detection of Loxosceles species (brown recluse) spider venom

Abstract

Study objective-Dermal lesions from unrelated arthropod species and medical causes appear similar to Loxosceles species (brown recluse spider) bites. This may result in delayed diagnosis and treatment. We developed a sensitive Loxosceles species venom enzyme-linked immunosorbent assay (ELISA) and characterized the specificity of the assay by evaluating antigenic cross-reactivity from a variety of North American arthropod venoms. Methods-NorthAmerican arthropod (14 spiders, 2 scorpions, and 1 bee) venoms were studied. Three venom amounts (diluted in 100 μL of ELISA buffer) were assayed: 16,000 ng, 2,000 ng, and 40 ng. The latter quantity was selected because this is the observed maximum amount of venom we detect when inoculating dermis with amounts likely to be deposited by a spider bite. The larger venom amounts are overwhelming quantities designed to test the limits of the assay for arthropod venom cross-reactivity. Similar amounts of Loxosceles species venom and bovine albumin served as positive and negative controls, respectively.Results-At the lowest amount of venom tested (40 ng), the ELISA detected only the Loxosceles species positive control. When 2,000 ng was assayed, only Scytodes fusca and Kukulcania hibernalis arachnid venoms (in addition to Loxosceles species) cross-reacted to the assay. Finally, at 16,000 ng, the ELISA assay modestly detected Diguetia canities, Heteropoda venatoria, Tegenaria agrestis, Plectreurys tristes, Dolomedes tenebrosus, and Hadrurus arizonensis arachnid venoms.Conclusion-Cross-reactivity was observed in 8 of 17 North American arthropod venoms when large venom amounts were assayed with a Loxosceles species ELISA. By using a relevant quantity of venom, 40 ng, the assay was specific for Loxosceles species venom. The venom specificity of the ELISA may allow clinical application in Loxosceles species endemic regions of North America.

Loxosceles: A Case Healed Completely Without Any Necrotic Tissue by Emergency Department and Review of the Literature

“…Günümüzde, spesifik bir tanı yöntemi ve türlerin ayırd edilebilmesi için Loxosceles olgularında çalışmalar devam etmektedir. Bu doğrultuda ELISA ile serumda Loxosceles zehrinin tespit edildiği, yine zehirle temastan 7 gün sonra tavşan modellerinin saç örneklerinde, yara yeri sürüntüsünde ve biyopsi materyalinde zehirin tespit edildiği çalışmalar yapılmıştır (14,15). Loxosceles ısırıkları genellikle ekstremitelerde görülmekle birlikte otorinolarinjoloji literatürüne geçecek şe-kilde kafa ve yüzde de lezyonlar bildirilmiştir (16).…”

Brown Recluse Spider Bite to the Face

“…7,9 According to Anderson, 1 the main criteria for diagnosing envenomation by a brown recluse should be that the spider is recovered in the immediate proximity of the bite and its species verified by an experienced entomologist. Gomez et al 10 identified a Loxosceles species venom enzyme-linked immunosorbent assay (ELISA) that accurately identified the venom at less than 40 ng/100 L homogenized rabbit tissue. At higher concentrations of venom, the assay cross-reacted with 8 of the 17 species tested.…”

“…Although not yet in clinical use, if applicable in humans this assay may prove to be valuable because it can at least identify the presence of an arthropod venom in the patient. 10 The venom is the etiologic agent responsible for the cutaneous necrosis. It is a low-molecular-weight, water-soluble protein expected to spread rapidly within the subcutaneous tissues.…”

Un posible patrón de oro para el diagnóstico de loxoscelismo y una solución para la investigación

“…4 , elaboraron un ensayo inmunoenzimático (ELISA) para la detección del veneno de las especies del género Loxosceles. Se estudió el veneno de diferentes artrópodos de Norteamérica (14 arañas, 2 escorpiones y una abeja).…”