Lopes C A, de Faria L S, de Sousa J E N, Borges I P, Ribeiro R P, Bueno L L, Rodrigues Ávila V M, Ferreira Júnior Á, Costa-Cruz J M
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil.
Laboratório de Bioquímica e Toxinas Animais, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil.
J Helminthol. 2019 Aug 14;94:e71. doi: 10.1017/S0022149X19000701.
Human ascariasis is a neglected tropical disease of great relevance to public health and is considered the most frequent helminthiasis in poor regions. Accurately diagnosing this parasite has been challenging due to limitations of current diagnostic methods. Immunoglobulin Y (IgY) technology is a very effective alternative for the production of highly specific and profitable antibodies. This study aimed to produce and apply anti-Ascaris suum IgY antibodies in the immunodiagnosis of human ascariasis. Five immunizations comprising total saline extract from A. suum adult life forms were given at 14-day intervals to Gallus gallus domesticus hens of the Isa Brown line. Eggs and blood samples were collected weekly and fortnightly, respectively, to monitor the production of antibodies. The specificity of antibodies was confirmed by dot-blot, kinetic enzyme-linked immunosorbent assay (ELISA), avidity ELISA, immunoblotting and indirect immunofluorescence antibody tests. The application for disease diagnosis was performed through the detection of immune complexes in human serum samples by sandwich ELISA. Peaks of IgY anti-A. suum production occurred at weeks 6 and 8. IgY showed high avidity levels after the second dose of immunization, ranging from 64% to 93%, with a mean avidity index of 78.30%. Purified IgY recognized 12 bands of proteins from A. suum saline extract. Eggs, the uterine portion and cuticles of A. suum female adult are reactive in immunofluorescence. The detection of immune complexes showed diagnostic values of 80% sensitivity and 90% specificity. In conclusion, specific IgY have been shown to be a potential immunodiagnostic tool with promising future applications in human ascariasis.
人体蛔虫病是一种被忽视的热带疾病,对公共卫生具有重大意义,被认为是贫困地区最常见的蠕虫病。由于现有诊断方法的局限性,准确诊断这种寄生虫一直具有挑战性。免疫球蛋白Y(IgY)技术是生产高度特异性和有益抗体的一种非常有效的替代方法。本研究旨在生产抗猪蛔虫IgY抗体并将其应用于人体蛔虫病的免疫诊断。以14天的间隔,用猪蛔虫成虫的总盐水提取物对伊莎褐系家鸡进行了五次免疫接种。每周和每两周分别采集鸡蛋和血液样本,以监测抗体的产生。通过斑点印迹、动力学酶联免疫吸附测定(ELISA)、亲和力ELISA、免疫印迹和间接免疫荧光抗体试验确认了抗体的特异性。通过夹心ELISA检测人血清样本中的免疫复合物来进行疾病诊断的应用。抗猪蛔虫IgY的产生高峰出现在第6周和第8周。第二次免疫接种后,IgY显示出高亲和力水平,范围为64%至93%,平均亲和力指数为78.30%。纯化的IgY识别出猪蛔虫盐水提取物中的12条蛋白带。猪蛔虫雌虫成虫的卵、子宫部分和表皮在免疫荧光中具有反应性。免疫复合物的检测显示诊断价值为80%的敏感性和90%的特异性。总之,特异性IgY已被证明是一种潜在的免疫诊断工具,在人体蛔虫病中具有广阔的未来应用前景。