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坦迪病毒糖蛋白Gn片段精细抗原表位的鉴定及ELISA检测方法的建立

Identification of fine antigenic epitopes of Tamdy virus glycoprotein Gn fragment and establishment of ELISA detection method.

作者信息

Fu Yujiao, Liu Liping, Zhang Beibei, Chao Xiaoshan, Jin Junxia, Wang Ying, Ding Juntao

机构信息

College of Life Science and Technology, Xinjiang University, Urumqi, 830017, China.

Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, Urumqi, 830017, China.

出版信息

Parasit Vectors. 2025 Jan 24;18(1):26. doi: 10.1186/s13071-024-06646-2.

DOI:10.1186/s13071-024-06646-2
PMID:39856717
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11762889/
Abstract

BACKGROUND

Tamdy virus (TAMV) was first isolated in Uzbekistan and Turkmenistan. In 2018, it was found in China, marking its entry into the molecular research era. TAMV is linked to febrile diseases, but its epidemiology and spillover risks are poorly understood, necessitating urgent molecular research and detection method development.

METHODS

The secondary structure of TAMV glycoprotein Gn was predicted, and the results showed that it had rich antigenic epitopes. According to the predicted results, glycoprotein Gn was divided into 46 truncated 16-peptides by modified synthetic peptide method, and the antigenicity of 46 truncated 16-peptides was verified by western blotting analysis.

RESULTS

The results showed that P8, P9, P24, P25, P28, P29, and P39 had antigenicity. Subsequently, the seven positive 16-peptide sequences with antigenicity were truncated to form 8-peptide sequences with an overlap of seven amino acids. After analysis with the same method, eight fine antigenic epitopes E1 (VINSTLDH), E2 (HVGSWGMP), E3 (SWGMPVTT), E4 (IRNQPFKS), E5 (FNVEVQ), E6 (AVVEHH), E7 (VEHHGNKA), and E8 (RGGRR) were identified, all of which were located on the three-dimensional surface of glycoprotein Gn and were highly conserved in different TAMV strains.

CONCLUSIONS

Eight precise epitopes were identified, and an indirect ELISA method based on fusion multiepitope peptide (r-Gn-MEPX) was developed and implemented, featuring high sensitivity, accuracy, and specificity.

摘要

背景

坦迪病毒（TAMV）最初在乌兹别克斯坦和土库曼斯坦被分离出来。2018年，在中国被发现，标志着其进入分子研究时代。TAMV与发热性疾病有关，但其流行病学和溢出风险了解甚少，因此迫切需要进行分子研究和开发检测方法。

方法

预测了TAMV糖蛋白Gn的二级结构，结果表明其具有丰富的抗原表位。根据预测结果，采用改良合成肽法将糖蛋白Gn分为46个截短的16肽，并通过蛋白质印迹分析验证了46个截短的16肽的抗原性。

结果

结果显示P8、P9、P24、P25、P28、P29和P39具有抗原性。随后，将七个具有抗原性的阳性16肽序列截短，形成重叠七个氨基酸的8肽序列。用相同方法分析后，鉴定出八个精细抗原表位E1（VINSTLDH）、E2（HVGSWGMP）、E3（SWGMPVTT）、E4（IRNQPFKS）、E5（FNVEVQ）、E6（AVVEHH）、E7（VEHHGNKA）和E8（RGGRR），它们均位于糖蛋白Gn的三维表面，且在不同TAMV毒株中高度保守。

结论

鉴定出八个精确表位，并开发并实施了基于融合多表位肽（r-Gn-MEPX）的间接ELISA方法，该方法具有高灵敏度、准确性和特异性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/8c57e327a9b2/13071_2024_6646_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/62b0c5c10fe4/13071_2024_6646_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/36b7d41da101/13071_2024_6646_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/181832ecbe4d/13071_2024_6646_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/fd20fab9cb26/13071_2024_6646_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/3f947587492a/13071_2024_6646_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/2e2642996cb9/13071_2024_6646_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/75ac3014e749/13071_2024_6646_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/11448ecfd75d/13071_2024_6646_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/8c57e327a9b2/13071_2024_6646_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/62b0c5c10fe4/13071_2024_6646_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/36b7d41da101/13071_2024_6646_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/181832ecbe4d/13071_2024_6646_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/fd20fab9cb26/13071_2024_6646_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/3f947587492a/13071_2024_6646_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/2e2642996cb9/13071_2024_6646_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/75ac3014e749/13071_2024_6646_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/11448ecfd75d/13071_2024_6646_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/734c/11762889/8c57e327a9b2/13071_2024_6646_Fig9_HTML.jpg

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