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评估种特异性多克隆抗体以检测和区分 和 。

Evaluation of species-specific polyclonal antibodies to detect and differentiate between and .

机构信息

Moredun Research Institute, Scotland, UK.

Royal (Dick) School of Veterinary Studies and the Roslin Institute, Scotland, UK.

出版信息

J Vet Diagn Invest. 2024 May;36(3):418-427. doi: 10.1177/10406387241234322. Epub 2024 Feb 29.

Abstract

Neosporosis and toxoplasmosis are major causes of abortion in livestock worldwide, leading to substantial economic losses. Detection tools are fundamental to the diagnosis and management of those diseases. Current immunohistochemistry (IHC) tests, using sera raised against whole parasite lysates, have not been able to distinguish between and We used and recombinant proteins, expressed in and purified using insoluble conditions, to produce specific polyclonal rabbit antisera. We aimed to develop species-specific sera that could be used in IHC on formalin-fixed, paraffin-embedded (FFPE) tissue sections to improve the diagnosis of ruminant abortions caused by protozoa. Two polyclonal rabbit sera, raised against recombinant proteins, anti--rNcSRS2 and anti--rTgSRS2, had specificity for the parasite they were raised against. We tested the specificity for each polyclonal serum using FFPE tissue sections known to be infected with and . The anti--rNcSRS2 serum labeled specifically only infected tissue blocks, and the anti--rTgSRS2 serum was specific to only infected tissues. Moreover, tissues from 52 cattle and 19 sheep previously diagnosed by lesion profiles were tested using IHC with our polyclonal sera and PCR. The overall agreement between IHC and PCR was 90.1% for both polyclonal anti-rNcSRS2 and anti-rTgSRS2 sera. The polyclonal antisera were specific and allowed visual confirmation of protozoan parasites by IHC, but they were not as sensitive as PCR testing.

摘要

刚地弓形虫病和新孢子虫病是全球家畜流产的主要原因,导致了巨大的经济损失。检测工具对于这些疾病的诊断和管理至关重要。目前,使用针对全寄生虫裂解物产生的血清进行的免疫组织化学(IHC)检测,无法区分 和 。我们使用 和 重组蛋白,在 和 条件下表达并通过不溶性条件进行纯化,产生了特异性的多克隆兔抗血清。我们旨在开发可用于固定在福尔马林中的石蜡包埋(FFPE)组织切片中的 IHC 的物种特异性血清,以改善对由原生动物引起的反刍动物流产的诊断。两种针对重组蛋白产生的多克隆兔血清,抗-rNcSRS2 和抗-rTgSRS2,对其产生的寄生虫具有特异性。我们使用已知感染 和 的 FFPE 组织切片测试了每种多克隆血清的特异性。抗-rNcSRS2 血清仅特异性标记感染组织块,而抗-rTgSRS2 血清仅特异性标记感染组织。此外,使用我们的多克隆血清和 PCR 对以前通过病变图谱诊断为 52 头牛和 19 只绵羊的组织进行了 IHC 检测。两种多克隆抗-rNcSRS2 和抗-rTgSRS2 血清的 IHC 和 PCR 之间的总体一致性为 90.1%。多克隆抗血清具有特异性,允许通过 IHC 可视化确认原生动物寄生虫,但它们的敏感性不如 PCR 检测。

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