【摘要】 目的 本研究建立了一种基于伽氏疏螺旋体(Borrelia garinii) SZ株全菌抗原的小鼠莱姆病血清学间接ELISA实验室诊断方法，为莱姆病的实验室诊断及流行病学监测提供标准化技术支撑。方法 通过系统优化实验条件，确定最佳抗原包被浓度、血清稀释比和酶标二抗浓度。制备小鼠感染模型，采集不同时间点血清，评估伽氏疏螺旋体与伯氏疏螺旋体B31株及阿氏疏螺旋体BO23株的交叉反应。结果 确定间接ELISA最佳反应体系为:抗原包被浓度0.2μg/μL、血清稀释比1∶200、酶标二抗浓度1∶2000。该方法在保持高灵敏度(OD值>0.8)的同时，显著降低了抗原用量。小鼠感染后12～25 d抗体水平达到峰值，特异性抗体比率(ABR%)阈值为41.7%，用于区分阳性和阴性样本。交叉反应分析表明，该方法与BO23株存在一定交叉，与B31株无明显交叉。结论 本研究成功建立了一种操作简便、经济高效的小鼠莱姆病间接ELISA诊断方法，适用于基层实验室及小规模筛查，为莱姆病的实验室诊断和流行病学监测提供了可靠的技术支持。更多还原

【Abstract】 Objective This study established an indirect enzyme-linked immunosorbent assay-based serological diagnostic method for Lyme disease in mice,using whole-cell antigens of Borrelia garinii strain SZ,providing standardized technical support for the laboratory diagnosis and epidemiological surveillance of Lyme disease.Methods Experimental conditions were systematically optimized to determine the optimal antigen coating concentration,serum dilution ratio,and enzyme-labeled secondary antibody concentration. A mouse infection model was established,and sera were collected at different time points to analyze antibody dynamics. Cross-reactivity with Borrelia burgdorferi strain B31 and Borrelia afzelii strain BO23 was evaluated. Results Through systematic optimization of experimental conditions, the optimal reaction system was determined to be an antigen coating concentration of 0. 2 μg/μL,serum dilution ratio of 1 ∶ 200,and enzyme-labeled secondary antibody concentration of1 ∶ 2000. This method ensured high sensitivity( optical density value > 0. 8) while significantly reducing antigen consumption. Antibody dynamic analysis revealed that serum antibody levels in infected mice peaked between 12 ～ 25 days post-inoculation,with a specificity antibody ratio threshold of 41. 7% to distinguish positive and negative samples. Cross-reactivity between Borrelia garinii and Borrelia burgdorferi( B31) as well as Borrelia afzelii( BO23)was also evaluated, indicating certain cross-reactivity with BO23 but no significant cross-reactivity with B31.Conclusions The result demonstrated that this method is simple,cost-effective,and suitable for primary laboratories and small-scale screening,providing a reliable standardized method for Lyme disease laboratory diagnosis and epidemiological surveillance.更多还原