Rikihisa Y, Ewing S A, Fox J C, Siregar A G, Pasaribu F H, Malole M B
Department of Veterinary Pathobiology, College of Veterinary Medicine, Ohio State University, Columbus 43210.
J Clin Microbiol. 1992 Jan;30(1):143-8. doi: 10.1128/jcm.30.1.143-148.1992.
Ehrlichia canis and canine granulocytic Ehrlichia sp. (CGE) infect canine monocytes and granulocytes, respectively. E. canis has been cultured in vitro and used to develop an immunofluorescence assay. CGE has not been cultured, and a serologic assay is not available. The sera of dogs infected with CGE were reported to react with E. canis by immunofluorescence. In this study, the temporal response of immunoglobulin G (IgG) was determined by an enzyme-linked immunosorbent assay (ELISA) with purified E. canis antigen in four dogs experimentally infected with E. canis, in two dogs experimentally infected with CGE, and in one dog infected with E. canis and subsequently infected with CGE. E. canis-infected dogs developed an IgG ELISA result of 1.5 or greater for the optical density signal/noise ratio by 2 months postinfection. CGE challenge of a dog with a previous E. canis infection induced an anamnestic increase in the IgG ELISA result; however, CGE infection alone did not induce a significant IgG ELISA response. Western immunoblot analysis showed that dogs infected with E. canis developed antibodies initially that reacted with low-molecular-mass proteins (30, 24, and 21 kDa) and subsequently with higher-molecular-mass proteins (160, 100, 78, 64, 47, and 40 kDa). In contrast, CGE-infected dogs showed reactions with the same higher-molecular-mass proteins of E. canis but, unlike E. canis-infected dogs, not with the low-molecular-mass proteins of E. canis. Of 10 serum samples collected in the field of Indonesia from dogs with tropical canine pancytopenia, all had an optical density signal minus noise value of 2.54 or greater in the IgG ELISA and reacted with E. canis antigen in a pattern similar to that of serum samples from dogs experimentally infected with E. canis in Western immunoblotting. This study suggests that the IgG ELISA and Western immunoblotting with purified E. canis as the antigen are useful in distinguishing between E. canis and CGE infections in dogs.
犬埃立克体和犬粒细胞埃立克体（CGE）分别感染犬的单核细胞和粒细胞。犬埃立克体已在体外培养，并用于开发免疫荧光测定法。CGE尚未培养成功，且尚无血清学检测方法。据报道，感染CGE的犬血清通过免疫荧光与犬埃立克体发生反应。在本研究中，采用酶联免疫吸附测定法（ELISA），用纯化的犬埃立克体抗原，测定了4只实验感染犬埃立克体的犬、2只实验感染CGE的犬以及1只先感染犬埃立克体随后又感染CGE的犬体内免疫球蛋白G（IgG）的时间反应。感染犬埃立克体的犬在感染后2个月时，IgG ELISA的光密度信号/噪声比结果达到1.5或更高。先前感染过犬埃立克体的犬受到CGE攻击后，IgG ELISA结果出现回忆性升高；然而，单独感染CGE并未诱导出显著的IgG ELISA反应。蛋白质免疫印迹分析表明，感染犬埃立克体的犬最初产生的抗体与低分子量蛋白质（30、24和21 kDa）反应，随后与高分子量蛋白质（160、100、78、64、47和40 kDa）反应。相比之下，感染CGE的犬与犬埃立克体相同的高分子量蛋白质发生反应，但与感染犬埃立克体的犬不同，它们不与犬埃立克体的低分子量蛋白质反应。在印度尼西亚热带犬全血细胞减少症犬的野外采集的10份血清样本中，所有样本在IgG ELISA中的光密度信号减去噪声值均为2.54或更高，并且在蛋白质免疫印迹中与犬埃立克体抗原的反应模式与实验感染犬埃立克体的犬的血清样本相似。本研究表明，以纯化的犬埃立克体为抗原的IgG ELISA和蛋白质免疫印迹法有助于区分犬感染犬埃立克体和CGE。
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