Institute of Animal Inspection and Quarantine, Chinese Academy of Inspection and Quarantine, 100176, Beijing, China.
People's Hospital of Jiaxiang, Jiaxiang County, Jining City, 272400, Shandong Province, China.
BMC Vet Res. 2021 Jan 23;17(1):48. doi: 10.1186/s12917-021-02760-w.
Brucellosis is a serious zoonosis disease that frequently causes significant economic loss in animal husbandry and threatens human health. Therefore, we established a rapid, accurate, simple and sensitive fluorescent immunochromatographic strip test (ICST) based on quantum dots (QDs) for detection the antibodies of Brucella infection animals serum.
The test strips were successfully prepared by quantum dot fluorescent microspheres (QDFM) as tracers, which were covalently coupled to an outer membrane protein of Brucella OMP22. The outer membrane protein OMP28 and monoclonal antibodies of OMP22 were separately dispensed onto a nitrocellulose membrane as test and quality control lines, respectively. The critical threshold for determining negative or positive through the ratio of the fluorescent signal of the test line and the control line (H / H) is 0.0492. The repeatability was excellent with an overall average CV of 8.78%. Under optimum conditions, the limit of detection was 1.05 ng/mL (1:512 dilution). With regard to the detection of brucellosis in 150 clinical samples, the total coincidence rate of ICST and Rose Bengal plate test (RBPT) was 97.3%, the coincidence rate of positive samples was 98.8%, the coincidence rate of negative samples was 95.3%, the sensitivity of RBPT is 1:32, and no cross reaction with the sera of other related diseases was observed.
In our present study, the QDFM has promising application for on-site screening of brucellosis owing to its high detection speed, high sensitivity, high specificity and low cost.
布鲁氏菌病是一种严重的人畜共患病，经常给畜牧业造成重大经济损失，并威胁人类健康。因此，我们建立了一种基于量子点（QDs）的快速、准确、简单、敏感的荧光免疫层析检测试纸条（ICST），用于检测布鲁氏菌感染动物血清中的抗体。
成功制备了量子点荧光微球（QDFM）作为示踪剂的测试条，该示踪剂通过共价键偶联到布鲁氏菌外膜蛋白 OMP22 上。将外膜蛋白 OMP28 和 OMP22 的单克隆抗体分别分配到硝酸纤维素膜上作为测试和质量控制线。通过测试线和控制线荧光信号的比值（H / H）来确定阴性或阳性的临界阈值为 0.0492。总体平均 CV 为 8.78%，重复性极好。在最佳条件下，检测限为 1.05ng/mL（1:512 稀释）。在对 150 份临床样本进行布鲁氏菌检测时，ICST 和虎红平板试验（RBPT）的总符合率为 97.3%，阳性样本的符合率为 98.8%，阴性样本的符合率为 95.3%，RBPT 的灵敏度为 1:32，并且与其他相关疾病的血清无交叉反应。
在本研究中，由于 QDFM 具有检测速度快、灵敏度高、特异性高、成本低等优点，有望用于现场布鲁氏菌病的筛查。
