Hill Dolores, Coss Cathleen, Dubey J P, Wroblewski Kristen, Sautter Mari, Hosten Tiffany, Muñoz-Zanzi Claudia, Mui Ernest, Withers Shawn, Boyer Kenneth, Hermes Gretchen, Coyne Jessica, Jagdis Frank, Burnett Andrew, McLeod Patrick, Morton Holmes, Robinson Donna, McLeod Rima
United States Department of Agriculture, Agricultural Research Service, Animal and Natural Resources Institute, Animal Parasitic Diseases Laboratory, Beltsville, Maryland 20705, USA.
J Parasitol. 2011 Apr;97(2):328-37. doi: 10.1645/GE-2782.1. Epub 2011 Mar 3.
Reduction of risk for human and food animal infection with Toxoplasma gondii is hampered by the lack of epidemiological data documenting the predominant routes of infection (oocyst vs. tissue cyst consumption) in horizontally transmitted toxoplasmosis. Existing serological assays can determine previous exposure to the parasite, but not the route of infection. We have used difference gel electrophoresis, in combination with tandem mass spectroscopy and Western blot, to identify a sporozoite-specific protein (T. gondii embryogenesis-related protein [TgERP]), which elicited antibody and differentiated oocyst- versus tissue cyst-induced infection in pigs and mice. The recombinant protein was selected from a cDNA library constructed from T. gondii sporozoites; this protein was used in Western blots and probed with sera from T. gondii -infected humans. Serum antibody to TgERP was detected in humans within 6-8 mo of initial oocyst-acquired infection. Of 163 individuals in the acute stage of infection (anti- T. gondii IgM detected in sera, or < 30 in the IgG avidity test), 103 (63.2%) had detectable antibodies that reacted with TgERP. Of 176 individuals with unknown infection route and in the chronic stage of infection (no anti- T. gondii IgM detected in sera, or > 30 in the IgG avidity test), antibody to TgERP was detected in 31 (17.6%). None of the 132 uninfected individuals tested had detectable antibody to TgERP. These data suggest that TgERP may be useful in detecting exposure to sporozoites in early T. gondii infection and implicates oocysts as the agent of infection.
由于缺乏流行病学数据来记录水平传播的弓形虫病的主要感染途径(摄入卵囊与组织包囊),降低人类和食用动物感染刚地弓形虫的风险受到阻碍。现有的血清学检测可以确定先前是否接触过该寄生虫,但无法确定感染途径。我们使用差异凝胶电泳,结合串联质谱和蛋白质印迹法,鉴定出一种子孢子特异性蛋白(刚地弓形虫胚胎发生相关蛋白 [TgERP]),该蛋白能引发抗体,并区分猪和小鼠中由卵囊与组织包囊引起的感染。重组蛋白是从由刚地弓形虫子孢子构建的cDNA文库中筛选出来的;该蛋白用于蛋白质印迹,并与刚地弓形虫感染人类的血清进行杂交。在初次获得卵囊感染后的6 - 8个月内,人类血清中检测到了针对TgERP的抗体。在163例处于感染急性期的个体(血清中检测到抗刚地弓形虫IgM,或IgG亲和力试验< 30)中,103例(63.2%)检测到与TgERP反应的抗体。在176例感染途径不明且处于感染慢性期的个体(血清中未检测到抗刚地弓形虫IgM,或IgG亲和力试验> 30)中,31例(17.6%)检测到针对TgERP的抗体。132例未感染个体的检测结果均未显示有可检测到的针对TgERP的抗体。这些数据表明,TgERP可能有助于检测刚地弓形虫早期感染中是否接触过子孢子,并提示卵囊是感染源。