[摘要]目的 初步建立一种脑脊液标本中常见病原菌的快速检测方法。方法 根据脑脊液标本中常见病原菌16S rRNA基因保守区的序列设计用于基因扩增的通用引物，制备细菌的通用探针、革兰阳性和阴性菌通用探针，以及肠杆菌科属、嗜血杆菌属和脑膜炎奈瑟菌、金黄色葡萄球菌、凝固酶阴性葡萄球菌、肺炎链球菌、产单核李斯特菌的属或种特异性探针，将探针加尾后固定于尼龙膜上制成寡核苷酸芯片。提取标本中病原菌DNA后用通用引物行PCR扩增，扩增过程中用生物素标记;将产物变性后分别与点有各种探针的尼龙膜反向杂交，检测脑脊液中细菌的感染情况。结果 所有探针均只与其相应的细菌DNA扩增产物反应产生杂交信号。结论 建立的脑脊夜病原菌寡核苷酸芯片能够准确、敏感、快速、高效地检测脑脊液标本中病原菌的感染情况。

    [关键词] 脑脊髓液;细菌;寡核苷酸芯片;寡核苷酸序列分析

    ESTABLISHMENT OF 16S rRNA OLIGO-CHIPS TO DETECT PATHOGENS IN CEREBROSPINAL FLUID

    BI CHUN-XIA， WU JING， ZHANG DE-KUN， et al

    (Department of Medical Laboratory， Qingdao Municipal Hospital， Qingdao 266011， China)

    [ABSTRACT]ObjectiveTo develop a rapid detection method for common pathogens in cerebrospinal fluid (CSF). Me- thodsAccording to the conservative and variable regions in bacterial 16S rRNA genes， a pair of universal primers that can amplify rDNA of all bacteria were designed. Ten kinds of specific probes were designed by Premier software. Probes for identification were tailed and then spotted onto a nylon membrane. DNA were isolated from each pathogen， and subjected to UP- PCR to amplify tar-get fragments ......