Savage H M, Duncan J F, Roberts D R, Sholdt L L
Department of Preventive Medicine and Biometrics, Uniformed Services University of the Health Sciences, Bethesda, MD 20814-4799.
J Am Mosq Control Assoc. 1991 Mar;7(1):16-23.
A dot-enzyme-linked immunosorbent assay, dot-ELISA, that allows for identification of human blood meals in mosquitoes in less than 2 h is presented. Strips of mylar-backed nitrocellulose paper, the dipstick, may be inoculated with anti-human capture antibody, blocked, dried and stored for at least 3 months before use. The Dipstick ELISA consistently detected human blood meals at 24 h post-blood meal in frozen Anopheles mosquitoes and at 32 h post-blood meal on samples eluted off filter-paper smears. This ELISA detects human IgG at dilutions of 1:25,600, and produces strong positive results at dilutions between 1:400 and 1:12,800. The Dipstick ELISA is highly specific, and no false positives were detected when tested against cow, horse, goat, dog, cat, pig, rabbit, mouse, rat, chicken, raccoon and opossum sera. All reagents for the assay are commercially available. The Dipstick ELISA meets requirements for a rapid and simple assay for the identification of human blood meals and should have particular application to short-term field studies and emergency epidemiological investigations. A modified protocol, the Trough ELISA, which treats dipsticks jointly in disposable pipet troughs during the conjugate and substrate steps, was developed and produced as a kit. The Trough ELISA produces 1-4% random false positives, and we recommend that the Trough ELISA not be used.
本文介绍了一种斑点酶联免疫吸附测定法（dot-ELISA），该方法可在不到2小时内鉴定蚊子体内的人血餐。聚酯薄膜背衬的硝酸纤维素纸条（试纸条）可用抗人捕获抗体接种，封闭，干燥，并在使用前储存至少3个月。试纸条酶联免疫吸附测定法在冷冻的按蚊体内血餐后24小时以及从滤纸条涂片上洗脱的样品血餐后32小时，均能持续检测到人血餐。这种酶联免疫吸附测定法能检测稀释至1:25,600的人IgG，并在1:400至1:12,800的稀释度下产生强阳性结果。试纸条酶联免疫吸附测定法具有高度特异性，在与牛、马、山羊、狗、猫、猪、兔、小鼠、大鼠、鸡、浣熊和负鼠血清进行检测时未检测到假阳性。该测定法的所有试剂均可从市场购得。试纸条酶联免疫吸附测定法符合快速简便鉴定人血餐的测定要求，尤其适用于短期现场研究和紧急流行病学调查。开发了一种改良方案——槽式酶联免疫吸附测定法（Trough ELISA），该方法在偶联物和底物步骤中在一次性移液管槽中联合处理试纸条，并制成试剂盒。槽式酶联免疫吸附测定法会产生1%-4%的随机假阳性，我们建议不要使用槽式酶联免疫吸附测定法。
