Research Institute for Tropical Medicine, 9002 Research Drive, Filinvest Corporate City, Alabang, Muntinlupa City, Philippines.
Research Institute for Tropical Medicine, 9002 Research Drive, Filinvest Corporate City, Alabang, Muntinlupa City, Philippines.
Acta Trop. 2020 Nov;211:105610. doi: 10.1016/j.actatropica.2020.105610. Epub 2020 Jun 28.
Rabies is a fatal zoonotic disease endemic in developing countries of Asia and Africa. Recently, the direct rapid immunohistochemical test (DRIT) was recommended by the World Health Organization (WHO) and the World Organization for Animal Health (OIE) as a diagnostic test for rabies. Therefore, a biotinylated polyclonal antibody (pAb) against the rabies lyssavirus (RABV) nucleoprotein was developed using a plasmid cDNA vaccine derived from a challenge virus standard 11 strain. A preliminary evaluation on the efficacy of this reagent in recognizing the Philippine RABV strain was tested using banked canine hippocampal tissue samples with DRIT and the results were compared to dFAT. The effects of acetone and formalin fixation on DRIT were also assessed through immunoreactivity scores of the specimens. Of the 142 samples examined, 104 tested positive and 38 negative using both dFAT and DRIT, showing 100% agreement between the two diagnostic procedures. Moreover, no false positive or false negative results were observed using acetone and formalin fixation. Thus, locally prepared biotinylated pAb from plasmid cDNA can be used for DRIT, especially in resource-limited laboratories in the Philippines. However, these results should be confirmed with a more thorough evaluation of this technique, and the range of detection needs to be further evaluated in a larger panel of animal samples and on other lyssaviruses.
狂犬病是一种在亚洲和非洲发展中国家流行的致命人畜共患病。最近,世界卫生组织(WHO)和世界动物卫生组织(OIE)推荐直接快速免疫组织化学检测(DRIT)作为狂犬病的诊断检测方法。因此,使用源自挑战病毒标准 11 株的质粒 cDNA 疫苗开发了针对狂犬病病毒核蛋白的生物素化多克隆抗体(pAb)。使用 DRIT 和 dFAT 测试了该试剂在识别菲律宾狂犬病病毒株方面的功效的初步评估,并将结果与 dFAT 进行了比较。还通过对标本的免疫反应评分评估了丙酮和福尔马林固定对 DRIT 的影响。在检查的 142 个样本中,使用 dFAT 和 DRIT 检测,104 个样本呈阳性,38 个样本呈阴性,两种诊断方法的一致性为 100%。此外,使用丙酮和福尔马林固定时,未观察到假阳性或假阴性结果。因此,可将从质粒 cDNA 制备的本地生物素化 pAb 用于 DRIT,尤其是在菲律宾资源有限的实验室中。但是,应该通过对该技术进行更彻底的评估来确认这些结果,并且需要在更大的动物样本组和其他狂犬病病毒上进一步评估检测范围。