基于核苷三磷酸二磷酸水解酶的马锥虫病间接ELISA抗体检测方法的建立

doi:10.3969/j.issn.1674-7968.2025.01.020

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Abstract Trypanosomosis (Surra) by Trypanosoma evansi, which resides in the blood, is a protozoan disease in animals such as horses (Equus) and camels (Camelus). Accurate diagnosis is a crucial step in the effective prevention and control of Surra, but the existing diagnostic techniques have many drawbacks such as high false negative rates and high prices. Based on recombinant expression of nucleoside triphosphate diphosphate hydrolase (TeNTPDase) from T. evansi, in this study, an ELISA method was established for Surra by optimizing various reaction parameters, including antigen coating concentration and conditions, serum dilution and incubation conditions, conjugated second antibody dilution and incubation conditions, and evaluated its diagnostic efficacy. The results showed that the recombinant TeNTPDase was mainly expressed in the form of inclusion bodies, with a size of approximately 60 kD. The coefficient of variation of the established ELISA method in different batches of coating antigens were 3.50%~7.32% being less than 10%, and the coefficient of variation at different incubation time points at 37 ℃ were 3.89%~14.45% being less than 15%, indicating good repeatability and stability. Moreover, when using the established ELISA method to detect clinical serum samples of horses, it was found that the positive rate of Surra was 5.83% (7/120), slightly higher than that of the commercial test kit (3.33%), but the difference was not significant, with a coincidence rate of 92.5% (111/120). This study provides technical support for the effective prevention and control of Surra in horses.

Key words： Surra Horse Nucleoside triphosphate diphosphohydrolase (NTPDase) ELISA

Received: 25 March 2024

ZTFLH:

S852.71

Corresponding Authors: * Corresponding authors: songhh@zafu.edu.cn; zhaoxiuling@customs.gov.cn

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	Articles by authors
	ZENG Qing-Qin
	LI Ru-Song
	LI Hong-Ying
	WANG Xin-Le
	WANG Ying
	XIA Tian-Qi
	WANG Pu
	SHI Heng-Zhi
	ZHANG Lu-Ping
	ZHENG Ya-Dong
	ZHAO Xiu-Ling
	SONG Hou-Hui

Cite this article:

ZENG Qing-Qin,LI Ru-Song,LI Hong-Ying, et al. Establishment of an Indirect ELISA Antibody Detection Method for Equine Trypanosomosis Based on Nucleotide Triphosphate Diphosphohydrolase[J]. 农业生物技术学报, 2025, 33(1): 220-227.

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https://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2025.01.020 OR https://journal05.magtech.org.cn/Jwk_ny/EN/Y2025/V33/I1/220

[1] 刘志强, 郭会玲, 金映红, 等. 2019. 新疆边境地区马、驼锥虫病流行病学调查[J]. 草食家畜, (01): 60-64.
(Liu Z Q, Guo H L, Jin Y H, et al. 2019. Epidemiological investigation on trypanosomosis in horses and camels in Xinjiang border region[J]. Grass-Feeding Livestock, (01): 60-64.)
[2] 王俊. 2019. 非洲锥虫病的研究进展[J].中国临床神经科学, 2019, 27(05): 547-554.
(Wang J.Research progress of African trypanosomiasis[J]. Chinese Journal of Clinical Neurosciences, 27(05): 547-554.)
[3] Desquesnes M, Gonzatti M, Sazmand A, et al.2022. A review on the diagnosis of animal trypanosomoses[J]. Parasites & Vectors, 15(1): 64.
[4] Gkeka A, Aresta-Branco, F Triller, et al.2023. Immunodominant surface epitopes power immune evasion in the African trypanosome[J]. Cell Reports, 42(3): 112262.
[5] González D A, Barbieri van Haaster M M, Quinteros Villarruel E, et al.2020. Salivary extracellular vesicles can modulate purinergic signalling in oral tissues by combined ectonucleoside triphosphate diphosphohydrolases and ecto-5'-nucleotidase activities[J]. Molecular and Cellular Biochemistry, 463(1-2): 1-11.
[6] Kim J, Álvarez-Rodríguez A, Li Z, et al.2023. Recent progress in the detection of Surra, a neglected disease caused by trypanosoma evansi with a one health impact in large parts of the tropic and sub-tropic world[J]. Microorganisms, 12(1): 44.
[7] Kumar R, Sethi K, Batra K, et al.2023. Exploring the potential of invariable surface glycoprotein (ISG65) as promising antigen for diagnosis of Trypanosoma evansi infection[J]. Veterinary Parasitology, 314: 109866.
[8] Mezzasoma L, Bellezza I, Orvietani P, et al.2022. Amniotic fluid stem cell-derived extracellular vesicles are independent metabolic units capable of modulating inflammasome activation in THP-1 cells[J]. FASEB Journal, 36(4): e22218.
[9] Paes-Vieira L, Gomes-Vieira A L, Meyer-Fernandes J R.2021. E-NTPDases: Possible roles on host-parasite interactions and therapeutic opportunities[J]. Frontiers in Cellular and Infection Microbiology, 11: 769922.
[10] Paula A T, Ribeiro K V G, Cardoso K F, et al.2022. Protective immunity triggered by ectonucleoside triphosphate diphosphohydrolase-based biopharmaceuticals attenuates cardiac parasitism and prevents mortality in Trypanosoma cruzi infection[J]. Bioorganic & Medicinal Chemistry, 72: 116966.
[11] Ribeiro I C, de Moraes J V B, Mariotini-Moura C, et al.2023. Synthesis of new non-natural L-glycosidic flavonoid derivatives and their evaluation as inhibitors of Trypanosoma cruzi ecto-nucleoside triphosphate diphosphohydrolase 1 (TcNTPDase1)[J]. Purinergic Signalling, 20(4):399-419.
[12] Sazmand A, Desquesnes M, Otranto D.2022. Trypanosoma evansi[J]. Trends in Parasitology, 38(6): 489-490.
[13] Truc P, Büscher P, Cuny G, et al.2013. Atypical human infections by animal trypanosomes[J]. PLOS Neglected Tropical Diseases, 7(9): e2256.
[14] Weiss P H, Batista F, Wagner G, et al.2015. Kinetic and biochemical characterization of Trypanosoma evansi nucleoside triphosphate diphosphohydrolase[J]. Experimental Parasitology, 153(2015): 98-104.
[15] Yadav S C, Kumar R, Kumar J, et al.2017. Antigenic characterization of 52-55 kDa protein isolated from Trypanosoma evansi and its application in detection of equine trypanosomosis[J]. Research in Veterinary Science, 114(2017): 455-460.
[16] Zheng L, Jiang N, Sang X, et al.2019. In-depth analysis of the genome of Trypanosoma evansi, an etiologic agent of Surra[J]. Science China. Life sciences, 62(3): 406-419.