Buffolano Wilma, Beghetto Elisa, Del Pezzo Mariassunta, Spadoni Andrea, Di Cristina Manlio, Petersen Eskild, Gargano Nicola
Kenton Laboratories, Via Pontina km 30.400, 00040 Rome, Italy.
J Clin Microbiol. 2005 Dec;43(12):5916-24. doi: 10.1128/JCM.43.12.5916-5924.2005.
The main objective of this work was to improve the early serologic diagnosis of toxoplasmosis in children at risk of congenital infection by using recombinant antigens. Serum samples from 104 infants born to mothers with primary Toxoplasma gondii infection acquired during pregnancy, of which 35 were congenitally infected and 22 had clinical silent toxoplasmosis at birth, were included. Immunoglobulin M (IgM), IgG, and IgG subtype antibodies against epitopes carried by fragments of T. gondii MIC2, MIC3, MIC4, M2AP, AMA1, and SAG1 gene products were measured by performing parallel enzyme immunoassays (Rec-ELISAs). Recombinant antigens preferentially reacted with IgG antibodies from infected infants compared to uninfected subjects (P < 0.0001), indicating that sera from infected children recognized a more diverse repertoire of antigens than sera transferred over the placenta from the mothers. Using two serial samples collected within 3 months of life, it was possible to demonstrate a neosynthesis of specific anti-MIC2 and anti-SAG1 immunoglobulin G, mainly of the IgG2 subtype, in 13 out of 20 infants with congenital toxoplasmosis. IgM antibodies in 97% of infected infants reacted with at least one of the recombinant antigens, confirming the diagnosis of congenital infection as soon as 2 months after birth (P < 0.0001). The use of recombinant antigens is effective in distinguishing T. gondii-infected from uninfected infants and shows that assays based on recombinant antigens improve the diagnosis of newborns with congenital toxoplasmosis.
这项工作的主要目标是通过使用重组抗原来改善对有先天性感染风险儿童的弓形虫病早期血清学诊断。研究纳入了104名母亲在孕期初次感染刚地弓形虫的婴儿的血清样本,其中35名婴儿先天性感染,22名在出生时患有临床无症状弓形虫病。通过进行平行酶免疫测定(重组酶联免疫吸附测定,Rec-ELISAs)来检测针对刚地弓形虫MIC2、MIC3、MIC4、M2AP、AMA1和SAG1基因产物片段所携带表位的免疫球蛋白M(IgM)、免疫球蛋白G(IgG)和IgG亚型抗体。与未感染的受试者相比,重组抗原优先与感染婴儿的IgG抗体发生反应(P < 0.0001),这表明感染儿童血清识别的抗原种类比通过胎盘从母亲转移过来的血清更多样化。使用在出生后3个月内采集的两份连续样本,在20名先天性弓形虫病婴儿中的13名中,有可能证明特异性抗MIC2和抗SAG1免疫球蛋白G主要是IgG2亚型的新生合成。97%感染婴儿的IgM抗体与至少一种重组抗原发生反应,在出生后2个月即可确诊先天性感染(P < 0.0001)。使用重组抗原可有效区分感染和未感染刚地弓形虫的婴儿,并表明基于重组抗原的检测方法可改善先天性弓形虫病新生儿的诊断。