Institute of Chemistry, University of Tartu, Ravila 14a, Tartu, 50 411, Estonia.
Institute of Chemistry, University of Tartu, Ravila 14a, Tartu, 50 411, Estonia; Tallinn University of Technology, Thomas Johann Seebeck Department of Electronics, Ehitajate tee 5, Tallinn, 12 618, Estonia.
Enzyme Microb Technol. 2019 Nov;130:109360. doi: 10.1016/j.enzmictec.2019.109360. Epub 2019 Jun 15.
Streptococcus uberis is a major mastitis-causing environmental pathogen, which rapid immunodetection has not been possible due to the absence of specific anti-Str. uberis antibodies. Recently, a specific antibody against the Str. uberis adhesion molecule (SUAM) has been designed. In the present study, the specificity and affinity of this antibody towards SUAM antigenic region SAPVYLGVSTE and Str. uberis cells are characterized, using experimental and in silico bioinformatic methods. The selectivity studies and bioinformatic analyses revealed high specificity of the antibody towards Str. uberis. The K value of SAPVYLGVSTE/anti-Str. uberis antibody complex was 27 ± 6 nM, indicating the applicability of this antibody for the detection of Str. uberis. The anti-Str. uberis antibody was used as a specific biorecognition element of a biosensor for the detection of Str. uberis bacteria in phosphate buffer and in milk and these analyses took less than 20 min. The Str. uberis biosensor was also tested in the milk of cows suffering from mastitis and the obtained results were in good agreement with the conventional identification of this pathogen by microbiological plating.
停乳链球菌是一种主要的乳腺炎环境病原体，由于缺乏特异性抗停乳链球菌抗体，因此无法快速进行免疫检测。最近，针对停乳链球菌黏附分子（SUAM）设计了一种特异性抗体。在本研究中，使用实验和计算生物信息学方法，对该抗体针对 SUAM 抗原区域 SAPVYLGVSTE 和停乳链球菌细胞的特异性和亲和力进行了表征。选择性研究和生物信息学分析表明，该抗体对停乳链球菌具有很高的特异性。SAPVYLGVSTE/抗停乳链球菌抗体复合物的 K 值为 27±6 nM，表明该抗体适用于检测停乳链球菌。抗停乳链球菌抗体被用作生物传感器的特异性生物识别元件，用于检测磷酸盐缓冲液和牛奶中的停乳链球菌细菌，这些分析耗时不到 20 分钟。停乳链球菌生物传感器还在患有乳腺炎的奶牛的牛奶中进行了测试，得到的结果与通过微生物平板培养法对该病原体的常规鉴定结果非常吻合。
