Serotyping of Toxoplasma gondii Infection Using Peptide Membrane Arrays
Serotyping of Toxoplasma gondii Infection Using Peptide Membrane Arrays
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DOI：
10.3389/fcimb.2019.00408
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发表时间：
2019-11-29
影响因子：
5.7
通讯作者：
Saeij, Jeroen P. J.
中科院分区：
文献类型：
--
作者：
Arranz-Solis, David;Cordeiro, Cynthia;Saeij, Jeroen P. J.
细胞内寄生虫弓形虫可以导致大多数温血动物的慢性感染，包括人类。在美国，通常分离出属于4种不同弓形虫克隆谱系（1、2、3和12型）的菌株，而不属于这些谱系的菌株在其他大陆（如南美洲）占主导地位。弓形虫株型在决定弓形虫感染的严重程度方面起着关键作用。因此，它是流行病学相关的开发一种非侵入性和廉价的方法来确定菌株类型在弓形虫感染和相关的基因型与疾病的结果。血清学分型是基于这样一个事实，即许多宿主抗体是针对免疫显性寄生虫蛋白，是高度多态性菌株之间。然而，目前的血清学检测只能可靠地区分2型和非2型感染。为了改善这些测定，小鼠，兔，和人感染血清反应对950肽从62个不同的多态性弓形虫蛋白，通过使用纤维素膜肽阵列。这使我们能够确定最抗原的肽，并查明最相关的多态性，确定菌株的特异性。我们的结果证实了先前描述的肽的效用，并鉴定了改善和增加测定的特异性的新型肽。此外，大量新的蛋白质显示出用于弓形虫诊断的潜力。其中，来自几个棒状体，致密颗粒，和表面蛋白的肽代表有前途的候选人，可用于在未来的实验中，以提高弓形虫血清分型。此外，已发表的GRA7分型肽的重新设计版本表现更好，并在小鼠，兔和人的血清中特异性区分3型与非3型感染。
The intracellular parasite Toxoplasma gondii can cause chronic infections in most warm-blooded animals, including humans. In the USA, strains belonging to four different Toxoplasma clonal lineages (types 1, 2, 3, and 12) are commonly isolated, whereas strains not belonging to these lineages are predominant in other continents such as South America. Strain type plays a pivotal role in determining the severity of Toxoplasma infection. Therefore, it is epidemiologically relevant to develop a non-invasive and inexpensive method for determining the strain type in Toxoplasma infections and to correlate the genotype with disease outcome. Serological typing is based on the fact that many host antibodies are raised against immunodominant parasite proteins that are highly polymorphic between strains. However, current serological assays can only reliably distinguish type 2 from non-type 2 infections. To improve these assays, mouse, rabbit, and human infection serum were reacted against 950 peptides from 62 different polymorphic Toxoplasma proteins by using cellulose membrane peptide arrays. This allowed us to identify the most antigenic peptides and to pinpoint the most relevant polymorphisms that determine strain specificity. Our results confirm the utility of previously described peptides and identify novel peptides that improve and increase the specificity of the assay. In addition, a large number of novel proteins showed potential to be used for Toxoplasma diagnosis. Among these, peptides derived from several rhoptry, dense granule, and surface proteins represented promising candidates that may be used in future experiments to improve Toxoplasma serotyping. Moreover, a redesigned version of the published GRA7 typing peptide performed better and specifically distinguished type 3 from non-type 3 infections in sera from mice, rabbits, and humans.