Identification and real-time expression analysis of selected Toxoplasma gondii in-vivo induced antigens recognized by IgG and IgM in sera of acute toxoplasmosis patients.
Identification and real-time expression analysis of selected Toxoplasma gondii in-vivo induced antigens recognized by IgG and IgM in sera of acute toxoplasmosis patients.
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DOI：
10.1186/1471-2334-13-287
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发表时间：
2013-06-24
影响因子：
3.7
通讯作者：
Noordin R
中科院分区：
文献类型：
--
作者：
Amerizadeh A;Khoo BY;Teh AY;Golkar M;Abdul Karim IZ;Osman S;Yunus MH;Noordin R
刚地弓形虫是一种专性细胞内的人畜共患寄生虫，属顶复门，可感染包括人类在内的多种温血动物。在这项研究中，体内诱导抗原的寄生虫进行了研究，使用体内诱导抗原技术（IVIAT）和合并血清从患者的血清学证据的急性感染。首先将合并的血清针对来自每种T的体外生长细胞的三种不同抗原制剂进行预吸收。gondii和E. coliXL 1-BlueMRF '，然后用其筛选T.弓形虫cDNA噬菌体表达文库对来自每组的阳性克隆进行体内和体外生长的寄生虫的mRNA的定量实时PCR表达分析。从IgM和IgG免疫筛选中，共发现29个反应性克隆与T.弓形虫基因实时荧光定量PCR表达分析显示，20个IgM检测基因和11个IgG检测基因相对于其体外表达水平在体内上调。这些基因包括微线体、固醇调节元件结合蛋白位点、SRS 34 A、MIC 2相关蛋白M2 AP、核氧还蛋白、蛋白磷酸酶2C和几种假设蛋白。IgG检测到的一种假设蛋白（GenBank登录号7899266）在体内与体外的倍数变化最大，为499.86;而IgM识别的另一种上调的假设蛋白（GenBank登录号7898829）在检测T.用20份个体血清样本检测弓形虫抗体。高度上调的基因和相应的蛋白质，特别是假设的蛋白质，可能有助于进一步研究了解疾病的发病机制，并作为潜在的疫苗候选人。
Toxoplasma gondii is an obligate intracellular zoonotic parasite of the phylum Apicomplexa which infects a wide range of warm-blooded animals, including humans. In this study in-vivo induced antigens of this parasite was investigated using in-vivo induced antigen technology (IVIAT) and pooled sera from patients with serological evidence of acute infection. The pooled sera was first pre-absorbed against three different preparations of antigens from in-vitro-grown cells of each T. gondii and E. coli XL1-Blue MRF’, subsequently it was used to screen T. gondii cDNA phage expression library. Positive clones from each group were subjected to quantitative real-time PCR expression analysis on mRNA of in-vivo and in-vitro grown parasites. A total of 29 reactive clones from each IgM and IgG immunoscreenings were found to have high homology to T. gondii genes. Quantitative real-time PCR expression analysis showed that 20 IgM-detected genes and 11 IgG-detected genes were up-regulated in-vivo relative to their expression levels in-vitro. These included genes encoding micronemes, sterol-regulatory element binding protein site, SRS34A, MIC2-associated protein M2AP, nucleoredoxin, protein phosphatase 2C and several hypothetical proteins. A hypothetical protein (GenBank accession no. 7899266) detected by IgG had the highest in-vivo over in-vitro fold change of 499.86; while another up-regulated hypothetical protein (GenBank accession no. 7898829) recognized by IgM showed high sensitivity (90%) and moderate specificity (70%) in detecting T. gondii antibodies when tested with 20 individual serum samples. The highly up-regulated genes and the corresponding proteins, in particular the hypothetical proteins, may be useful in further studies on understanding the disease pathogenesis and as potential vaccine candidates.
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影响因子：
2.5
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
3.1
作者：
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通讯作者：
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影响因子：
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作者：
El Hajj, Hiba;Papoin, Julien;Lebrun, Maryse
通讯作者：
Lebrun, Maryse