School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, 310013, China.
Engineering Research Center of Novel Vaccine of Zhejiang Province, Hangzhou Medical College, Hangzhou, China.
Parasit Vectors. 2024 Feb 22;17(1):81. doi: 10.1186/s13071-024-06180-1.
Timely diagnosis of Toxoplasma gondii infection is necessary to prevent and control toxoplasmosis transmission. The gold immunochromatographic assay (GICA) is a means of rapidly detecting pathogen in samples. GICA-based diagnostic methods have been developed to accurately detect pathogens with high sensitivity and specificity, and their application in T. gondii diagnosis is expected to yield good results.
Colloidal gold test strips were produced using T. gondii C-terminal truncated apical membrane antigen 1 (AMA1C). Colloidal gold-AMA1C and colloidal gold-murine protein conjugate were synthesized under optimal conditions. A nitrocellulose membrane was treated with AMA1C and goat anti-mouse antibody as the test line and control line, respectively. In total, 90 cat serum samples were tested using AMA1C-GICA and a commercial enzyme linked immunosorbent assay (ELISA) kit. The GICA results were digitally displayed using a portable colloidal gold immunochromatographic test strip analyzer (HMREADER). The sensitivity, specificity, and stability of AMA1C-GICA were assessed, and this was then used to examine clinical samples, including 203 human sera, 266 cat sera, and 81 dog sera.
AMA1C-GICA had a detection threshold of 1:32 for T. gondii-positive serum. The GICA strips specifically detected T. gondii antibodies and exhibited no reactivity with Plasmodium vivax, Paragonimus kellicotti, Schistosoma japonicum, Clonorchis sinensis, and Schistosoma mansoni. Consequently, 15 (16.7%) positive samples were detected using the AMA1C-GICA and commercial ELISA kits for each of the assays. The receiver-operating characteristic curve showed that GICA had a relative sensitivity of 85.3% and specificity of 92%, with an area under the curve of 98%. After analyzing clinical samples using HMREADER, 1.2%-23.4% of these samples were found to be positive for T. gondii.
This study presents a novel assay that enables timely and efficient detection of serum antibodies against T. gondii, thereby allowing for its early clinical diagnosis. Furthermore, the integration of digital detection using HMREADER can enhance the implementation of GICA.
及时诊断刚地弓形虫感染对于预防和控制弓形虫病的传播至关重要。金免疫层析法（GICA）是一种快速检测样品中病原体的方法。已经开发了基于 GICA 的诊断方法，以高灵敏度和特异性准确检测病原体，其在弓形虫诊断中的应用有望取得良好的效果。
使用刚地弓形虫 C 末端截短的顶膜抗原 1（AMA1C）制备胶体金测试条。在最佳条件下合成胶体金-AMA1C 和胶体金-鼠蛋白缀合物。硝酸纤维素膜分别用 AMA1C 和山羊抗鼠抗体处理作为测试线和控制线。共检测了 90 份猫血清样品，使用 AMA1C-GICA 和商业酶联免疫吸附试验（ELISA）试剂盒进行检测。使用便携式胶体金免疫层析测试条分析仪（HMREADER）对 GICA 结果进行数字显示。评估了 AMA1C-GICA 的灵敏度、特异性和稳定性，并用于检测临床样本，包括 203 份人血清、266 份猫血清和 81 份狗血清。
AMA1C-GICA 对刚地弓形虫阳性血清的检测阈值为 1:32。GICA 条特异性检测刚地弓形虫抗体，与间日疟原虫、卫氏并殖吸虫、日本血吸虫、华支睾吸虫和曼氏血吸虫无反应。因此，使用 AMA1C-GICA 和商业 ELISA 试剂盒对每种检测方法均检测到 15（16.7%）个阳性样本。受试者工作特征曲线显示，GICA 的相对灵敏度为 85.3%，特异性为 92%，曲线下面积为 98%。使用 HMREADER 分析临床样本后，发现这些样本中有 1.2%-23.4%对刚地弓形虫呈阳性。
本研究提出了一种新的检测方法，能够及时有效地检测血清中抗刚地弓形虫的抗体，从而实现其早期临床诊断。此外，使用 HMREADER 进行数字检测的整合可以增强 GICA 的实施。
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