Abstract: Objective　To explore a new technique of rapid diagnosis of purulent meningitis in neonates. Methods　Cerebrospinal fluid（ CSF ） specimens from 24 cases of suspected purulent meningitis were detected by 16SrRNA Microarrays in the Children’s Hospital affilated to Zhejiang University from August 2003 to February，2006； the bacterial cultures of CSF were done as controls at the same time. Results　（1） The PCR amplified products of 49 strains were tested by the gene chip hybridization， including 33 Gram-positve and 16 Gram-negative strains. Using the amplified DNA of the 33 Gram-positive bacteria as templets， there were positive findings for the two universal probes and the one Gram-positve probe. The Gram-negative probes and universal probes showed positive results when 16 Gram-negative bacteria were used as templates. Each strain specifically hybridized with its own probe. （2） 1pg（10-12g）（10 copies）were detected by 16S rRNA gene chip hybridization at least in the PCR amplified products of Εscherichia coli-enteric bacterium， equaled to 2 bacteria nearly. （3） Of the 24 cases，11 were positive by 16S rRNA Microarrays， the positive rate 45.83% （11/24） being significantly higher than that of CSF culture （12.50%） （3/24） （P < 0.01）. Conclusion　16SrRNA Microarrays with higher specificity and sensitivity， demanding less CSF， is rapid and reliable for diagnosing purulent meningitis in neonates. So it has considerable value of application.

Key words: purulent meningitis, CSF , gene chip, hybridization

摘要： 目的　探讨新生儿化脓性脑膜炎（以下简称化脑）新的快速诊断方法。 方法　2003年8至2006年2月浙江大学医学院附属儿童医院采用脑脊液细菌16S rRNA 基因芯片技术对24例临床上疑似化脑患儿脑脊液（CSF）的细菌DNA进行测定，同期进行与CSF细菌培养的对照。 结果　（1）49株细菌PCR扩增产物基因芯 片杂交结果显示，33株G+菌其G+探针和通用探针均阳性，G-探针阴性；16株G-菌中，G-探针和通用探针均阳性，G+探针阴性，均能与相应特异的探针发生杂交。 （2）大肠埃希菌DNA的PCR产物其16S rRNA 基因芯片的最小检出量为1 pg，约等于10个拷贝数，相当于2个细菌。 （3）16S rRNA 基因芯片检测24份脑脊液标本 发现11份阳性，阳性率为45.83% （11/24），明显高于脑脊液培养的阳性率12.50% （3/24），差异有统计学意义（ P < 0.01）。 结论　脑脊液细菌16S rRNA 基因芯片检测技术特异性强、敏感性高，需标本量少，是早期快速诊断儿童化脑的可靠方法，具有较大的应用价值。

关键词: 16SrRNA, 基因芯片, 杂交, 化脓性脑膜炎, 脑脊液