Graczyk T K, Cranfield M R, Shiff C J
Johns Hopkins University, School of Hygiene and Public Health, Department of Immunology and Infectious Diseases, Baltimore, Maryland 21205.
J Parasitol. 1994 Oct;80(5):713-8.
Erythrocytic stages of Haemoproteus columbae were extracted from the cytoplasm of nucleated red blood cells (RBC) of Rock dove pigeons (Columba livia) using cationic detergent (N,N',N'-polyoxyethylene(10)-N-tallow-1,3-diaminopropane [EDTA-20]) and discontinuous Percoll gradient density. Crude RBC extract (CRBCE) antigen was prepared. Parasitized RBCs were more resistant to EDTA-20 action than unparasitized cells. An enzyme-linked immunosorbent assay (ELISA) was developed for detection of anti-H. columbae immunoglobulins in 30 wild-captured C. livia. Whole blood, serum, and dried blood on filter paper gave similar results; the latter was selected for sampling convenience. Optimal antigen concentration was 5 micrograms/ml, and anti-H. columbae immunoglobulins were detectable at a 10(-4.11) dilutions. The binding efficacy of anti-chicken IgG to the pigeon immunoglobulins was significantly higher than anti-duck IgG or anti-turkey IgG. Parasitemia by Giemsa-stained thin blood smears ranged from 20.0 to 47.5%, mean = 32.4 +/- 8.3%; 17 of 30 birds had multiply infected RBCs with a mean parasitemia of 2.4 +/- 1.1%, range 0.7-4.9%; 27 birds were positive by the ELISA. No clinical signs of infection were observed. ELISA absorbance values were not correlated with the level of parasitemia in individual birds. All pigeons were negative for anti-Plasmodium relictum and anti-P. elongatum immunoglobulins as determined by ELISA. The pigeons were not subclinically infected with Plasmodium spp. as determined by inoculation of domestic ducklings with blood from dexamethasone-immunosuppressed pigeons.
使用阳离子去污剂（N,N',N'-聚氧乙烯(10)-N-牛脂-1,3-二氨基丙烷[EDTA-20]）和不连续的Percoll梯度密度，从岩鸽（Columba livia）有核红细胞（RBC）的细胞质中提取鸽血变原虫的红细胞阶段。制备了粗红细胞提取物（CRBCE）抗原。被寄生的红细胞比未被寄生的细胞对EDTA-20的作用更具抗性。开发了一种酶联免疫吸附测定（ELISA）法，用于检测30只野生捕获的岩鸽体内的抗鸽血变原虫免疫球蛋白。全血、血清和滤纸上的干血给出了相似的结果；由于采样方便，选择了后者。最佳抗原浓度为5微克/毫升，在10(-4.11)稀释度下可检测到抗鸽血变原虫免疫球蛋白。抗鸡IgG与鸽免疫球蛋白的结合效力显著高于抗鸭IgG或抗火鸡IgG。经吉姆萨染色的薄血涂片检测的寄生虫血症范围为20.0%至47.5%，平均值 = 32.4 +/- 8.3%；30只鸟中有17只的红细胞被多重感染，平均寄生虫血症为2.4 +/- 1.1%，范围为0.7 - 4.9%；27只鸟通过ELISA检测呈阳性。未观察到感染的临床症状。ELISA吸光度值与个体鸟的寄生虫血症水平无关。通过ELISA测定，所有鸽子的抗残疟原虫和抗伸长疟原虫免疫球蛋白均为阴性。通过用来自地塞米松免疫抑制鸽子的血液接种家鸭雏，确定鸽子未被疟原虫属亚临床感染。
