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应用荧光偏振分析检测动物布鲁氏菌病,使用荧光标记的合成寡糖作为生物传感示踪剂。

Applying a Fluorescence Polarization Assay for Detection of Brucellosis in Animals Using the Fluorescently Labeled Synthetic Oligosaccharides as Biosensing Tracer.

机构信息

Department of Chemistry, Moscow State University, 119991 Moscow, Russia.

N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 119991 Moscow, Russia.

出版信息

Biosensors (Basel). 2024 Aug 21;14(8):404. doi: 10.3390/bios14080404.

Abstract

Brucellosis in animals is an infectious disease caused by bacteria of the genus . Known methods for diagnosing brucellosis face some challenges, due to the difficulties in isolating and standardizing the natural brucellosis antigen. In this work, we investigated the possibility of using the fluorescence polarization assay (FPA) with synthetic glycoconjugate biosensing tracers to detect antibodies against as a new methodology for diagnosing brucellosis. Based on the received results, the synthetic fluorescein-labeled trisaccharide tracer is most effective for Brucellosis detection. This tracer is structurally related to the immune determinant fragment of the LPS buildup of N-formyl-d-perosamine units, connected via α-(1→3)-linkage at the non-reducing end and α-(1→2)-linkage at the reducing end. The sensitivity and specificity in the case of the use of trisaccharide tracer were 71% and 100% (Yuden's method) and 87% and 88% (Euclidean method), respectively, which is comparable with the diagnostic efficiency of traditionally used serological methods, such as the agglutination test (AT), complement fixation test (CFT), and Rose Bengal test (RBT). Given the known advantages of FPA (e.g., speed, compactness of the equipment, and standard reagents) and the increased specificity of the developed test system, it would be appropriate to consider its widespread use for the diagnosis of brucellosis in animals, including rapid testing in the field.

摘要

动物布鲁氏菌病是一种由 属细菌引起的传染病。已知的布鲁氏菌病诊断方法面临一些挑战,因为难以分离和标准化天然布鲁氏菌抗原。在这项工作中,我们研究了使用荧光偏振测定法(FPA)与合成糖缀合物生物传感示踪剂检测针对 的抗体作为一种新的布鲁氏菌病诊断方法的可能性。基于所得到的结果,合成荧光素标记的三糖示踪剂在布鲁氏菌病检测中最有效。该示踪剂与 N-甲酰-d-高丝氨酸单位 LPS 堆积的免疫决定簇片段结构相关,通过非还原端的α-(1→3)-键和还原端的α-(1→2)-键连接。使用三糖示踪剂的情况下,灵敏度和特异性分别为 71%和 100%(Yuden 法)和 87%和 88%(欧几里得法),与传统使用的血清学方法(如凝集试验(AT)、补体结合试验(CFT)和孟加拉玫瑰红试验(RBT))的诊断效率相当。鉴于 FPA 的已知优势(例如,速度、设备的紧凑性和标准试剂)和开发的测试系统的特异性提高,考虑广泛用于动物布鲁氏菌病的诊断,包括现场快速测试是合适的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e0f/11352515/9434c601b0d1/biosensors-14-00404-g001.jpg

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