Analysis of clonal type-specific antibody reactions in Toxoplasma gondii seropositive humans from Germany by peptide-microarray.
Analysis of clonal type-specific antibody reactions in Toxoplasma gondii seropositive humans from Germany by peptide-microarray.
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DOI：
10.1371/journal.pone.0034212
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Schares G
中科院分区：
文献类型：
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作者：
Maksimov P;Zerweck J;Maksimov A;Hotop A;Gross U;Spekker K;Däubener W;Werdermann S;Niederstrasser O;Petri E;Mertens M;Ulrich RG;Conraths FJ;Schares G
不同克隆型的刚地弓形虫被认为与不同的感染临床表现有关。血清分型是一种新技术，可用于确定人类感染的弓形虫克隆型，并将分型研究扩展到更大的人群，包括感染但未患病的个体。利用54个先前发表的模拟克隆型特异性表位的合成肽，建立了一种用于弓形虫血清分型的肽微阵列检测方法。该试验应用于急性弓形虫感染（21例）、潜伏性弓形虫感染（53例）和弓形虫血清阳性的森林工人（100例）采集的人类血清（174例）。大多数（n = 124; 71%）弓形虫血清阳性的人血清对克隆II型特异性合成肽（II型肽）有反应。I型和III型多肽分别被42% （n = 73）和16% （n = 28）的人血清识别，II-III型、I - III型和I - ii型多肽分别被49% （n = 85）、36% （n = 62）和14% （n = 25）的人血清识别。模拟ii型特异性表位的合成肽的反应强度最高。一部分血清（n = 22; 13%）与类型特异性肽无反应。与潜伏弓形虫感染或血清阳性的森林工人相比，急性弓形虫病患者的多肽数量有统计学意义上的显著增加。ii型特异性反应在研究人群中比例过高且强度较高，这与先前在同一地区进行的弓形虫卵囊基因分型研究一致。也有个体有I型或iii型特异性反应。需要充分表征的参考血清和进一步的特异性肽标记物来建立和执行未来更高分辨率的血清分型方法。
Different clonal types of Toxoplasma gondii are thought to be associated with distinct clinical manifestations of infections. Serotyping is a novel technique which may allow to determine the clonal type of T. gondii humans are infected with and to extend typing studies to larger populations which include infected but non-diseased individuals. A peptide-microarray test for T. gondii serotyping was established with 54 previously published synthetic peptides, which mimic clonal type-specific epitopes. The test was applied to human sera (n = 174) collected from individuals with an acute T. gondii infection (n = 21), a latent T. gondii infection (n = 53) and from T. gondii-seropositive forest workers (n = 100). The majority (n = 124; 71%) of all T. gondii seropositive human sera showed reactions against synthetic peptides with sequences specific for clonal type II (type II peptides). Type I and type III peptides were recognized by 42% (n = 73) or 16% (n = 28) of the human sera, respectively, while type II–III, type I–III or type I–II peptides were recognized by 49% (n = 85), 36% (n = 62) or 14% (n = 25) of the sera, respectively. Highest reaction intensities were observed with synthetic peptides mimicking type II-specific epitopes. A proportion of the sera (n = 22; 13%) showed no reaction with type-specific peptides. Individuals with acute toxoplasmosis reacted with a statistically significantly higher number of peptides as compared to individuals with latent T. gondii infection or seropositive forest workers. Type II-specific reactions were overrepresented and higher in intensity in the study population, which was in accord with genotyping studies on T. gondii oocysts previously conducted in the same area. There were also individuals with type I- or type III-specific reactions. Well-characterized reference sera and further specific peptide markers are needed to establish and to perform future serotyping approaches with higher resolution.