检测HSV等5种病原体抗体的微阵列技术  

Study on detecting antibodies of five pathogens using microarray technique

作　　者：刘毅[1,2] 曹鲁全 黄海燕[1] 王进鸿 韩金祥[1] 

机构地区：[1]山东省医药生物技术研究中心,国家卫生部生物技术药物重点实验室,山东省现代医用药物与技术重点实验室,济南250062 [2]济南市立四院 [3]济南市妇幼保健院 [4]山东省胸科医院

出　　处：《中华微生物学和免疫学杂志》2006年第8期756-760,共5页Chinese Journal of Microbiology and Immunology

基　　金：山东省重点攻关项目基金资助(2004GG2202150)

摘　　要：目的建立抗原微阵列技术检测多种病原体抗体。方法将病原体的基因工程抗原,以电脑操纵的机械手将其点在赖氨酸修饰的玻片上,制备抗原微阵列,与血清反应后随之与Cy3标记的二抗反应,用激光共聚焦扫描仪扫描玻片,检测血清中相应病原体的IgG,并进行了灵敏度和重复性的检测。结果5种抗原的最低检测限度为HSV1-gG 1．56μg／ml,HSV2-gG 3．125μg/ml,CMV- p150 1．56μg/ml,RV-E1E2 3．125μg／ml,MT-LAM 31．26μg/ml;抗原微阵列与ELISA检测的结果相比有较高的符合率;检测IgG和IgM的灵敏度分别为50 pg、10 Pg;不同批次2张玻片间总体变异系数为IgG 6．52,IgM 18．89。结论抗原微阵列可用于平行检测血液中病原体特异性抗体。Objective To set up antigen microarray technique for detecting multiple antibodies of pathogens. Methods The reconbinant antigens of pathogens were printed on lysine- modified glass slides with a computer-controlled robots. The slides were incubated with serum samples and subsequendy reacted with Cy3 labeled secondary antibodies. The antibodies of specific pathogens in serum were detected and scanned using a laser confocal scanner. The sensitivity and repeatability of the method were detected. Results The lowest detection limits of HSV1-gG, HSV2-gG, CMV-p150, RV-E1E2, TB-LAM were 1.56 μg/ml, 3. 125 μg/ml, 1.56 μg/ml, 3. 125 μg/ ml, and 31.26 μg/ml respectively. There were good concordance between microarray assay and ELISA in the assay of sera. The sensitivities of IgG and IgM were 50 pg and 10 pg respectively. The CV between two chips was IgG 6.52, IgM 18.89. Conclusion The antigen microarray has potential advantages for detection of specific pathogen antibodies.

关 键 词：抗原微阵列 病原体 抗体 

分 类 号：R446.6[医药卫生—诊断学]