Thongpoo Preeyanuch, Methawiroon Jiravich, Mangkit Bandid, Rucksaken Rucksak, Sussadee Metita, Rangubpit Warin, Unajak Sasimanas, Jittapalapong Sathaporn, Suwan Eukote
Program of Science and Mathematics (Biology), Faculty of Science and Technology, Phuket Rajabhat University, Phuket 83000, Thailand.
Department of Veterinary Technology, Faculty of Veterinary Technology, Kasetsart University, Bangkok 10900, Thailand.
Animals (Basel). 2025 Jun 26;15(13):1893. doi: 10.3390/ani15131893.
, a pathogen of significant concern in animal production, companion animal health, and public health, particularly affects immunocompromised individuals and pregnant women. Current diagnostic techniques employ both direct and indirect methods, with serological assays widely used for detecting infections in humans and animals. In this study, the TIM-barrel structure of Br2 -glucosidase was engineered to create 10 chimeric multi-epitope proteins for serological detection. Indirect ELISA screening identified three promising candidate proteins, V4Z, SFF, and S7V-V4Z-SFF, with sensitivities ranging from 71-86% and specificities ranging from 68-76%. Among these, ELISA-V4Z achieved the highest concordance with the reference IFAT method (Kappa = 0.58, 95% CI = 0.32-0.84) and demonstrated a moderate positive predictive value (PPV, 67%) and strong negative predictive value (NPV, 90%). These results suggest that the V4Z chimeric protein demonstrated the strongest performance among the tested candidates for detection, exhibiting the highest sensitivity and specificity along with moderate agreement with the reference IFAT. However, its overall diagnostic performance remains limited. These findings highlight the need for further refinement and validation to enhance its diagnostic potential and assess its applicability for broader serological testing.
作为动物生产、伴侣动物健康和公共卫生领域备受关注的病原体，尤其会影响免疫功能低下的个体和孕妇。目前的诊断技术采用直接和间接方法，血清学检测广泛用于检测人和动物的感染。在本研究中，对Br2 -葡萄糖苷酶的TIM桶状结构进行工程改造，以创建10种嵌合多表位蛋白用于血清学检测。间接ELISA筛选鉴定出三种有前景的候选蛋白，V4Z、SFF和S7V-V4Z-SFF，其灵敏度范围为71-86%，特异性范围为68-76%。其中，ELISA-V4Z与参考IFAT方法的一致性最高（Kappa = 0.58，95% CI = 0.32-0.84），并显示出中等阳性预测值（PPV，67%）和强阴性预测值（NPV，90%）。这些结果表明，V4Z嵌合蛋白在检测的候选物中表现最强，具有最高的灵敏度和特异性，与参考IFAT方法有中等一致性。然而，其整体诊断性能仍然有限。这些发现强调需要进一步优化和验证，以提高其诊断潜力并评估其在更广泛血清学检测中的适用性。
