School of Life Sciences, Zhengzhou Universitygrid.207374.5, Zhengzhou, People's Republic of China.
Microbiol Spectr. 2022 Oct 26;10(5):e0142922. doi: 10.1128/spectrum.01429-22. Epub 2022 Sep 26.
Bluetongue (BT) is a severe noncontagious infectious disease that occurs in sheep and wild ruminants but occasionally also in cattle and camels. The worldwide BT pandemic has had a significant impact on global livestock production. Rapid detection helps prevent outbreaks of bluetongue disease. Fluorescence-linked immunosorbent assay (FLISA) labeled with quantum dots (QDs) is typically used for detection due to its high sensitivity. There has been no reported detection of BT virus (BTV) using QD-based fluorescence immunoassays. In this study, monoclonal antibodies (MAbs) against BT were prepared by immunizing BALB/c mice with recombinant VP7 protein. Two MAbs with high sensitivity and specificity were selected as the detection antibody (2F11) and capture antibody (11B7). Then, the detection antibody was coupled with QDs to prepare QD-MAb fluorescence probes. Fluorescence-linked immunosorbent assay is highly specific, detecting only VP7 protein/BTV, and did not show any nonspecific reactions with other reoviruses. The detection limit of VP7 protein was 3.91 ng/mL using fluorescence-linked immunosorbent assay, with a coefficient of variation (CV) of less than 15%. The establishment of rapid, sensitive direct FLISA has potential for bluetongue virus detection and control of BT vaccine quality. Bluetongue virus causes the severe infectious disease BT. BTV has many serotypes, and there is no cross-protection among different serotypes. BT is listed as a notifiable animal infectious disease by the World Organisation for Animal Health (OIE) and occurs throughout the world, causing significant economic losses. The establishment of a fast and effective detection method is the key to controlling and preventing this disease. Current methods for detecting BTV mainly include reverse transcription-PCR (RT-PCR), enzyme-linked immunosorbent assays (ELISA), and immunochromatographic strips that are based on antigen-antibody recognition. Immunoassays are most commonly used because of their low cost, high specificity, and fast analysis, making them particularly useful for routine monitoring. These conventional detection strategies for BTV have some drawbacks. Recently, FLISA has been drawing attention due to its sensitivity, which is higher than traditional immunoassays. Fluorescence-linked immunosorbent assays (FLISA) using fluorescent materials as labels overcome ELISA's disadvantage of being time-consuming.
蓝舌病（BT）是一种严重的非传染性传染病，发生在绵羊和野生反刍动物中，但偶尔也发生在牛和骆驼中。全球蓝舌病大流行对全球畜牧业生产产生了重大影响。快速检测有助于预防蓝舌病的爆发。荧光酶联免疫吸附试验（FLISA）标记量子点（QDs）通常用于检测，因为其灵敏度高。尚未有报道使用基于 QD 的荧光免疫分析检测 BT 病毒（BTV）。在这项研究中，通过用重组 VP7 蛋白免疫 BALB/c 小鼠制备了针对 BT 的单克隆抗体（MAbs）。选择了两种具有高灵敏度和特异性的 MAbs 作为检测抗体（2F11）和捕获抗体（11B7）。然后，将检测抗体与 QDs 偶联以制备 QD-MAb 荧光探针。荧光酶联免疫吸附试验具有高度特异性，仅检测 VP7 蛋白/BTV，与其他呼肠孤病毒无任何非特异性反应。荧光酶联免疫吸附试验检测 VP7 蛋白的灵敏度检测限为 3.91ng/mL，变异系数（CV）小于 15%。快速、灵敏的直接 FLISA 的建立有可能用于蓝舌病病毒的检测和 BT 疫苗质量的控制。蓝舌病病毒引起严重的传染病 BT。BTV 有许多血清型，不同血清型之间没有交叉保护。蓝舌病被世界动物卫生组织（OIE）列为法定动物传染病，在世界各地发生，造成重大经济损失。建立快速有效的检测方法是控制和预防这种疾病的关键。目前检测 BTV 的方法主要包括逆转录-PCR（RT-PCR）、酶联免疫吸附试验（ELISA）和基于抗原-抗体识别的免疫层析条，其中免疫分析因其成本低、特异性高、分析速度快而最常用，使其特别适用于常规监测。这些传统的 BTV 检测策略存在一些缺点。最近，由于其灵敏度高于传统免疫分析，FLISA 引起了人们的关注。使用荧光材料作为标记的荧光酶联免疫吸附试验（FLISA）克服了 ELISA 耗时的缺点。
