Marinkelle C J, Vallejo G A, Schottelius J, Guhl F, de Sanchez N
Acta Trop. 1986 Sep;43(3):215-23.
Flagellates of Trypanosoma cruzi (stock Molino 1), obtained from the intestine of experimentally infected Rhodnius prolixus, grown in cellular or acellular culture, as well as from the blood of infected mice, were examined by a direct fluorescence test using the lectins RCA (Ricinus communis-120) and SBA (soy bean agglutinin; Glycine maxima), conjugated with fluorescein isothiocyanate, for the detection of beta-D-galactose and alpha,beta-N-acetyl-D-galactosamine on the membranes of the flagellates. The same reactions were carried out using Trypanosoma rangeli (stock San Agustin), obtained from the intestine, hemo-lymph or salivary glands of experimentally infected R. prolixus, as well as from cultures and from the blood of experimentally infected CFW mice. The results indicate that the membrane of T. rangeli in the salivary glands of the vector contains beta-D-galactose, but that this sugar is absent from all other developmental stages of this trypanosome. All stages of intestinal and cultured. T. cruzi presented positive reactions with RCA-FITC and SBA-FITC. The high specificity of this technique makes it useful for the examination of R. prolixus, previously used in xenodiagnosis of Chagas' disease and for the examination of intradomiciliary or sylvatic vectors in epidemiological surveys in areas where T. cruzi and T. rangeli coexist. Formaldehyde fixed samples can be examined months later and false reports due to T. rangeli can be avoided.
对从实验感染的克氏锥蝽肠道中获取的克氏锥虫(莫利诺1株)鞭毛虫进行检测,这些鞭毛虫在细胞或无细胞培养基中培养,也取自感染小鼠的血液,采用直接荧光试验,使用与异硫氰酸荧光素偶联的凝集素RCA(蓖麻凝集素-120)和SBA(大豆凝集素;大豆),检测鞭毛虫膜上的β-D-半乳糖和α,β-N-乙酰-D-半乳糖胺。对从实验感染的克氏锥蝽肠道、血淋巴或唾液腺中获取的,以及来自培养物和实验感染CFW小鼠血液中的兰氏锥虫(圣奥古斯丁株)进行同样的反应。结果表明,在媒介唾液腺中的兰氏锥虫膜含有β-D-半乳糖,但在该锥虫的所有其他发育阶段均不存在这种糖。肠道和培养的克氏锥虫的所有阶段对RCA-FITC和SBA-FITC均呈阳性反应。该技术的高特异性使其可用于检测先前用于恰加斯病异体诊断的克氏锥蝽,以及在克氏锥虫和兰氏锥虫共存地区的流行病学调查中检测家栖或野生媒介。甲醛固定的样本数月后仍可检测,且可避免因兰氏锥虫导致的假报告。