Myint K S, Linthicum K J, Tanskul P, Lerdthusnee K, Vaughn D W, Manomuth C, Mongkolsirichaikul D, Hansukjariya P, Hastriter M W
Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand.
J Med Entomol. 1998 Jul;35(4):556-60. doi: 10.1093/jmedent/35.4.556.
Immunocytochemical methods were developed and tested for their ability to detect the distribution of Orientia tsutsugamushi in paraffin sections of adult chiggers (Leptotrombidium imphalum Vercammen-Grandjean & Langston). Rickettsial antigen was detected by application of a simple direct or amplified immunocytochemistry procedure and an indirect immunofluorescent procedure. In the direct procedure alkaline phosphatase conjugation to the mouse polyclonal antibody to the Karp strain was followed by the HistoMark Red test system to detect rickettsial antigen. The amplification procedure used a similar method but used an unlabeled primary antibody followed by secondary biotinylated antimouse IgG, streptavidin-alkaline phosphatase, and the HistoMark Red test system. The immunofluorescent procedure included a biotinylated secondary antibody followed by addition of a streptavidin-FITC conjugate. Specific tissue tropisms in infected chiggers were observed in the salivary glands, nervous tissue, and ovaries of adult female mites in all procedures; however, nonspecific fluorescence of the chigger limited definitive identification of tissue tropisms with the indirect immunofluorescent procedure.
开发并测试了免疫细胞化学方法，以检测恙虫病东方体在成年恙螨（地里纤恙螨Vercammen-Grandjean & Langston）石蜡切片中的分布。通过应用简单的直接或放大免疫细胞化学程序以及间接免疫荧光程序来检测立克次体抗原。在直接程序中，将碱性磷酸酶与针对Karp株的小鼠多克隆抗体结合，然后使用HistoMark Red检测系统来检测立克次体抗原。放大程序使用类似方法，但使用未标记的一抗，随后是生物素化的抗小鼠IgG二抗、链霉亲和素-碱性磷酸酶以及HistoMark Red检测系统。免疫荧光程序包括生物素化二抗，随后添加链霉亲和素-FITC共轭物。在所有程序中，在成年雌性螨的唾液腺、神经组织和卵巢中均观察到受感染恙螨的特定组织嗜性；然而，恙螨的非特异性荧光限制了通过间接免疫荧光程序对组织嗜性的明确鉴定。
