Development of an Immunochromatographic Assay Based on Dense Granule Protein 7 for Serological Detection of Toxoplasma gondii Infection
Development of an Immunochromatographic Assay Based on Dense Granule Protein 7 for Serological Detection of Toxoplasma gondii Infection
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DOI：
10.1128/cvi.00747-12
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发表时间：
2013-04-01
影响因子：
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通讯作者：
Nishikawa, Yoshifumi
中科院分区：
文献类型：
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作者：
Terkawi, Mohamad Alaa;Kameyama, Kyohko;Nishikawa, Yoshifumi
弓形虫致密颗粒抗原蛋白（TgGRAs）是一种潜在的诊断抗原。TgGRA7和TgGRA14在T.提示TgGRAs可能是高抗原性蛋白。在此，将TgGRA7和TgGRA14作为开发用于快速诊断测试的标记物的候选物进行评价。采用间接酶联免疫吸附试验（iELISA）比较了TgGRA7和TgGRA14重组蛋白的特异性和敏感性。实验感染小鼠，并使用重组T.弓形虫主要表面抗原2（TgSAG2）作为参考对照。用TgGRA7的iELISA显示出最高的诊断准确性，并且可以在急性和慢性感染中检测抗TgGRA7抗体。同时对59份现场采集的猪样品进行了检测，并与乳胶凝集试验（LAT）进行了比较。在3种重组抗原中，TgGRA7的阳性率最高，与LAT结果的一致性为88.14，kappa值为0.76。此外，开发了基于重组TgGRA7的免疫层析试验（ICT），用于快速检测感染的抗体。ICT与T.感染的小鼠和未感染的或新孢子虫感染的小鼠。用ICT检测猪血清，结果与LAT和iELISA的TgGRA7结果相比有利，kappa值分别为0.66和0.70至0.79。这些数据表明，基于TgGRA7的ICT是一种很有前途的诊断工具，可用于临床常规检测和现场样本的大规模筛选。
Dense granule antigen proteins derived from Toxoplasma gondii (TgGRAs) are potential antigens for the development of diagnostic tools. TgGRA7 and TgGRA14 were detected in the peritoneal fluid of T. gondii-infected mice, suggesting that TgGRAs may be highly antigenic proteins. Here, TgGRA7 and TgGRA14 were evaluated as candidates for the development of a marker for a rapid diagnostic test. The specificity and sensitivity of purified recombinant proteins of TgGRA7 and TgGRA14 were compared in an indirect enzyme-linked immunosorbent assay (iELISA) using a series of serum samples from T. gondii-experimentally infected mice and using recombinant T. gondii major surface antigen 2 (TgSAG2) as a reference control. The iELISA with TgGRA7 showed the greatest diagnostic accuracy and could detect anti-TgGRA7 antibody in acute and chronic infections. A total of 59 field samples from pigs were also examined by the iELISAs, and the results compared with those of the latex agglutination test (LAT). Among the three recombinant antigens, TgGRA7 had the highest rates of positivity, with significant concordance (88.14) and kappa value (0.76) in comparison with the results using LAT. Furthermore, an immunochromatographic test (ICT) based on recombinant TgGRA7 was developed for rapid detection of antibodies to the infection. The ICT differentiated clearly between sera from T. gondii-infected mice and uninfected or Neospora caninum-infected mice. Pig sera were examined with the ICT, and the results compared favorably with those of LAT and iELISA for TgGRA7, with kappa values of 0.66 and 0.70 to 0.79, respectively. These data suggest that the ICT based on TgGRA7 is a promising diagnostic tool for routine testing in the clinic and mass screening of samples in the field.