Gomez Hernan F, Krywko Diann M, Stoecker William V
Department of Emergency Medicine, University of Michigan Medical Center, Ann Arbor, MI 48109-0305, USA.
Ann Emerg Med. 2002 May;39(5):469-74. doi: 10.1067/mem.2002.122914.
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.
North American arthropod (14 spiders, 2 scorpions, and 1 bee) venoms were studied. Three venom amounts (diluted in 100 microL 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.
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.
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.
来自无关节肢动物物种和医学病因的皮肤损伤看起来与棕色遁蛛叮咬相似。这可能导致诊断和治疗延迟。我们开发了一种灵敏的棕色遁蛛属毒液酶联免疫吸附测定（ELISA），并通过评估多种北美节肢动物毒液的抗原交叉反应来表征该测定的特异性。
对北美节肢动物（14种蜘蛛、2种蝎子和1种蜜蜂）的毒液进行了研究。测定了三种毒液量（用100微升ELISA缓冲液稀释）：16000纳克、2000纳克和40纳克。选择后者的量是因为这是我们在用可能由蜘蛛叮咬沉积的量接种真皮时检测到的毒液最大量。较大的毒液量是为了测试该测定对节肢动物毒液交叉反应的限度而设置的过量。相似量的棕色遁蛛属毒液和牛血清白蛋白分别作为阳性和阴性对照。
在测试的最低毒液量（40纳克）时，ELISA仅检测到棕色遁蛛属阳性对照。当测定2000纳克时，只有褐足塞托蛛和冬栖库库卡尼亚蛛的蛛形纲动物毒液（除棕色遁蛛属外）与该测定发生交叉反应。最后，在16000纳克时，ELISA适度检测到苍白迪盖蛛、狩猎巨蟹蛛、田野褛网蛛、暗色栉足蛛、阴暗盗蛛和亚利桑那肥尾蝎的蛛形纲动物毒液。
当用棕色遁蛛属ELISA测定大量毒液时，在17种北美节肢动物毒液中的8种中观察到了交叉反应。通过使用相关量的毒液（40纳克），该测定对棕色遁蛛属毒液具有特异性。ELISA的毒液特异性可能使其在北美棕色遁蛛属流行地区具有临床应用价值。
