Assay for Detection of Loxosceles Envenomation

斜索蛇毒的检测测定

• 批准号: 8101097
• 负责人: WILLIAM V STOECKER
• 金额: $ 39.14万
• 依托单位: STOECKER & ASSOCIATES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8101097
• 关键词: Accident and Emergency department; Address; Affinity; American; Antibodies; Antivenins; Area; Biological Assay; Bioreactors; Biotinylation; Bite; Buffers; Cessation of life; Clinical; Clinical Research; Color; Contracts; Country; Cutaneous; Detection; Development; Device Approval; Devices; Diagnosis; Diagnostic; Diagnostic tests; Differential Diagnosis; Economics; Enzyme-Linked Immunosorbent Assay; Evaluation; Feedback; Friends; Fright; Goals; Health; Hearing; Hour; Human; In Vitro; Infectious Skin Diseases; Inflammatory; Label; Lateral; Legal patent; Lesion; Life; Manufacturer Name; Marketing; Measures; Mediation; Methods; Missouri; Modification; Monoclonal Antibodies; Morbidity - disease rate; Necrosis; Oryctolagus cuniculus; Pain; Painless; Patients; Phase; Physicians; Proteins; Protocols documentation; Relative (related person); Research; Seasons; Sensitivity and Specificity; Site; Skin; Skin Cancer; Source; Specificity; Spider Bites; Spider Venoms; Spiders; Surface; Surveys; Swab; Techniques; Testing; Time; Triage; Tube; United States; Venoms; Work; assay development; clinical application; commercialization; experience; in vivo; loxosceles venom; mortality; phase 1 study; polyclonal antibody; satisfaction; sphingomyelin phosphodiesterase D; tool; wound

DESCRIPTION (provided by applicant): Envenomations by the brown recluse spider, Loxosceles reclusa, are a significant source of morbidity in endemic regions of the United States, and misdiagnoses are common. A survey of physicians in the endemic area has shown the economic viability of an accurate diagnostic test for these spider bites. Development and testing of an optimized Loxosceles venom assay will present significant challenges. Unlike the routine construction of ELISAs dedicated to the detection of a single protein, this ELISA will detect venom containing multiple proteins, including a unique physiologically active protein- sphingomyelinase D (SMD) abundantly present in the venom. In preliminary research, our polyclonal assay has shown good sensitivity and good in-vitro specificity. Our research shows that identifiable amounts of venom in clinical envenomations are present for at least seven days. In rabbits, our polyclonal assay allows identification of venom on the surface for as long as two weeks. The limits of sensitivity, in-vivo specificity, and the duration of detection are unknown. Phase I should allow determination of the smallest amount of venom detectable as well as the clinical time limits for in-vivo duration of sensitivity and specificity of the assay. Monoclonal antibodies will be isotyped and quantities raised in a bioreactor to perform checkerboard analysis. This analysis will allow determination of an optimal combination of L. reclusa monoclonal and polyclonal antibodies. A kit assay will result from Phase I, allowing multi-site testing in Phase II. These clinical studies will allow development of a lateral flow assay or microtiter plate assay, with the goal of FDA device approval and marketing. Clinical application of an optimized assay would save the morbidity and expense due to inappropriate diagnosis and treatment of various skin conditions with presentations similar to Loxosceles envenomations. Techniques used in the successful detection of this spider venom are directly applicable to bites from S. American Loxosceles species, responsible for additional deaths each year. The swab venom assay technique could be applicable to envenomations from numerous species PUBLIC HEALTH RELEVANCE: Bites of the brown recluse spider, Loxosceles reclusa, cause considerable morbidity and occasional mortality in the Midwest. Many lesions can appear similar and mimic spider bites, including bacterial and fungal skin infections as well as skin cancer. The goal of this project is to develop a commercially viable test for brown recluse spider bites using a painless and simple swab test for the spider venom on the surface of the skin.

描述（由申请人提供）：由褐隐蛛（Loxosceles reclusa）所引起的中毒是美国一些地方病区的一个重要致病源，误诊是很常见的。对流行地区的医生进行的一项调查显示，对这些蜘蛛叮咬进行准确的诊断测试具有经济可行性。开发和测试优化的Loxosceles毒液分析将面临重大挑战。与常规构建的用于检测单一蛋白质的ELISA不同，该ELISA将检测含有多种蛋白质的毒液，包括一种独特的生理活性蛋白-鞘磷脂酶D (SMD)，该蛋白大量存在于毒液中。在初步研究中，我们的多克隆检测方法具有良好的灵敏度和体外特异性。我们的研究表明，在临床中毒中，可识别数量的毒液至少存在7天。在兔子身上，我们的多克隆试验允许在长达两周的时间内识别表面的毒液。其灵敏度、体内特异性和检测时间的限制尚不清楚。第一阶段应允许测定可检测到的最小毒液量，以及测定体内敏感性和特异性持续时间的临床时间限制。单克隆抗体将被同型并在生物反应器中提高数量以进行棋盘分析。该分析将允许确定一个最佳组合的单克隆和多克隆抗体。第一阶段将产生试剂盒分析，第二阶段将进行多地点测试。这些临床研究将允许开发横向流动测定法或微量滴度板测定法，目标是FDA设备批准和营销。临床应用一种优化的检测方法，将节省由于不适当的诊断和治疗各种与蛇形囊肿相似的皮肤病而导致的发病率和费用。用于成功检测这种蜘蛛毒液的技术直接适用于南美洲Loxosceles物种的叮咬，每年造成额外的死亡。拭子毒液检测技术可适用于多种物种的毒液检测

项目成果

Duration of Loxosceles reclusa venom detection by ELISA from swabs.

通过 ELISA 从拭子检测 Loxosceles reclusa 毒液的持续时间。

• 发表时间: 2009
• 期刊: Clinical laboratory science : journal of the American Society for Medical Technology
• 作者: McGlasson,DavidL;Green,JonathonA;Stoecker,WilliamV;Babcock,JamesL;Calcara,DavidA
• 通讯作者: Calcara,DavidA

Systemic loxoscelism confirmation by bite-site skin surface: ELISA.

通过咬伤部位皮肤表面确认系统性异侧运动：ELISA。

• 发表时间: 2009
• 期刊: Missouri medicine
• 作者: Stoecker,WilliamV;Wasserman,GaryS;Calcara,DavidA;Green,JonathanA;Larkin,Karen
• 通讯作者: Larkin,Karen