Brown M, Shami Y, Zywulko M, Singh-Naz N, Middleton P J
Department of Microbiology, University of Toronto, Ontario, Canada.
J Clin Microbiol. 1990 Jun;28(6):1398-402. doi: 10.1128/jcm.28.6.1398-1402.1990.
A time-resolved fluoroimmunoassay (TR FIA) was developed for the direct detection of adenovirus types 40 (Ad40) and 41 (Ad41) in stool specimens by using a monoclonal antibody (5D8/2C2) which recognizes both Ad40 and Ad41 but does not cross-react with other adenovirus serotypes. In this assay, the detector antibody is biotinylated directly rather than labeled with europium, and the fluorescent signal is generated on a solid phase in the presence of excess europium (Eu3+). The strength of the signal is dependent on the amount of a Eu3+ chelator [4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA)]-streptavidin complex bound by the biotinylated detector antibody (5D8/2C2). In a pilot study with 41 specimens, this TR FIA demonstrated a maximum sensitivity and specificity of 88% compared with SmaI restriction analysis of adenovirus isolates from the same specimens. TR FIA using the europium chelator BCPDA represents a feasible approach for the direct identification of specific adenovirus serotypes in stool specimens.
开发了一种时间分辨荧光免疫测定法（TR FIA），用于通过使用一种单克隆抗体（5D8/2C2）直接检测粪便标本中的40型腺病毒（Ad40）和41型腺病毒（Ad41），该抗体可识别Ad40和Ad41，但不与其他腺病毒血清型发生交叉反应。在该测定法中，检测抗体直接用生物素化而非用铕标记，并且在存在过量铕（Eu3+）的情况下在固相上产生荧光信号。信号强度取决于由生物素化检测抗体（5D8/2C2）结合的Eu3+螯合剂[4,7-双（氯磺苯基）-1,10-菲咯啉-2,9-二羧酸（BCPDA）]-链霉亲和素复合物的量。在对41份标本的初步研究中，与对来自相同标本的腺病毒分离株进行SmaI限制性分析相比，该TR FIA显示出的最大灵敏度和特异性为88%。使用铕螯合剂BCPDA的TR FIA是直接鉴定粪便标本中特定腺病毒血清型的一种可行方法。
