A novel recombinant Theileria annulata surface protein as an antigen in indirect enzyme-linked immunosorbent assay for the serological diagnosis of tropical theileriosis.

IF 1.7 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Veterinary World Pub Date : 2024-08-01 Epub Date: 2024-08-27 DOI:10.14202/vetworld.2024.1936-1942
Anara Ryskeldina, Aleksandr Korobeinikov, Nailya Tursunbay, Maxat Berdikulov, Alexandr Shevtsov, Christian Bauer, Yersyn Mukhanbetkaliyev, Marat Kuibagarov
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Abstract

Background and aim: Theileria annulata infection in cattle causes major economic losses in livestock production in many Central Asian countries, including the southern region of Kazakhstan. This study aimed to obtain a recombinant T. annulata surface protein (TaSP) and to investigate its possible use as an antigen in an indirect enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of bovine theileriosis.

Materials and methods: Recombinant TaSP was obtained by cloning a polymorphic region of the TaSP gene, expressing it in Escherichia coli strain BL21, and purifying it by metal chelating chromatography. An indirect ELISA using recombinant TaSP as an antigen was developed and evaluated for the detection of T. annulata-specific antibodies in plasma samples from 69 cows polymerase chain reaction (PCR)-positive or PCR-negative for T. annulata and/or Theileria orientalis from southern Kazakhstan.

Results: The obtained recombinant protein had a molecular weight of 32 kDa, and mass spectrometry analysis of the purified protein identified it as a fragment of the surface protein of T. annulata. Initial testing of 69 field plasma samples from cattle showed that the results of indirect ELISA using TaSP as an antigen agreed substantially with those of T. annulata PCR (κ: 0.78). The relative sensitivity and specificity of indirect ELISA were 88.7% and 100%, respectively, using PCR as a reference. There was no evidence of cross-reaction with T. orientalis.

Conclusion: Initial results using recombinant TaSP as an antigen in indirect ELISA are promising and support the widespread use of this assay for routine diagnosis and T. annulata seroprevalence studies in cattle in Kazakhstan and possibly neighboring countries.

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一种新的重组环状毛癣菌表面蛋白,作为间接酶联免疫吸附试验的抗原,用于热带毛癣菌病的血清学诊断。
背景和目的:在包括哈萨克斯坦南部地区在内的许多中亚国家,牛感染环状丝虫病给畜牧业生产造成了重大经济损失。本研究旨在获得重组环状丝虫病表面蛋白(TaSP),并研究其在间接酶联免疫吸附试验(ELISA)中作为抗原用于牛丝虫病血清学诊断的可能性:通过克隆 TaSP 基因的多态区,在大肠杆菌 BL21 菌株中表达并用金属螯合层析法纯化,获得重组 TaSP。以重组 TaSP 为抗原,开发并评估了间接 ELISA 法,用于检测哈萨克斯坦南部 69 头聚合酶链式反应(PCR)阳性或阴性环状丝虫病和/或东方丝虫病奶牛血浆样本中的环状丝虫病特异性抗体:结果:获得的重组蛋白分子量为 32 kDa,对纯化蛋白进行的质谱分析确定其为环斑蓟马表面蛋白的一个片段。对 69 份牛的野外血浆样本进行的初步检测表明,以 TaSP 为抗原的间接 ELISA 检测结果与环斑驳虫 PCR 检测结果基本一致(κ:0.78)。以 PCR 为参照,间接 ELISA 的相对灵敏度和特异性分别为 88.7% 和 100%。没有证据表明与 T. orientalis 发生了交叉反应:在间接 ELISA 中使用重组 TaSP 作为抗原的初步结果很有希望,支持在哈萨克斯坦和可能的邻国的牛群中广泛使用该检测方法进行常规诊断和环斑蓟马血清流行率研究。
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来源期刊
Veterinary World
Veterinary World Multiple-
CiteScore
3.60
自引率
12.50%
发文量
317
审稿时长
16 weeks
期刊介绍: Veterinary World publishes high quality papers focusing on Veterinary and Animal Science. The fields of study are bacteriology, parasitology, pathology, virology, immunology, mycology, public health, biotechnology, meat science, fish diseases, nutrition, gynecology, genetics, wildlife, laboratory animals, animal models of human infections, prion diseases and epidemiology. Studies on zoonotic and emerging infections are highly appreciated. Review articles are highly appreciated. All articles published by Veterinary World are made freely and permanently accessible online. All articles to Veterinary World are posted online immediately as they are ready for publication.
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