中国比较医学杂志2026，Vol.36Issue(3)：93-99,7.DOI:10.3969/j.issn.1671-7856.2026.03.008
小鼠莱姆病血清学间接ELISA实验室诊断方法的建立
Establishment of an indirect enzyme linked immunosorbent assay for serological diagnosis of Lyme disease in mice
摘要
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 of 1∶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.关键词
莱姆病/间接ELISA/小鼠模型/伽氏疏螺旋体/血清学诊断Key words
Lyme disease/indirect ELISA/mouse model/Borrelia garinii/serological diagnosis分类
医药卫生引用本文复制引用
翟斌涛,包碧波,马少霞,李甲,周雅馨,李冰,张继瑜..小鼠莱姆病血清学间接ELISA实验室诊断方法的建立[J].中国比较医学杂志,2026,36(3):93-99,7.基金项目
陇原青年创新创业人才项目(2024QNTD41) （2024QNTD41）
甘肃省自然科学基金(23JRRA562) （23JRRA562）
中国农业科学院创新工程(CAAS-ASTIP-2014-LIHPS) （CAAS-ASTIP-2014-LIHPS）
国家肉牛牦牛产业技术体系项目(CARS-37) （CARS-37）
兰州市青年科技人才创新项目(2023-QN-73). （2023-QN-73）
