HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Free Full Text (PDF) Free Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Liu, Y. Articles by Zhu, B. Search for Related Content PubMed PubMed Citation Articles by Liu, Y. Articles by Zhu, B.

---

ORIGINAL RESEARCH ARTICLE

Development and Evaluation of 16S rDNA Microarray for Detecting Bacterial Pathogens in Cerebrospinal Fluid

Yi Liu^*,, Jin-Xiang Han^*,,1, Hai-Yan Huang^* and Bo Zhu^*

^* Key Laboratory for Biotech-Drugs Ministry of Health, Shandong Medicinal Biotechnology Center, Jinan 250062, China; and Medical College of Shandong University, Jinan 250012, China

To whom requests for reprints should be addressed at ¹ Shandong Medicinal Biotechnology Center, Jinan 250062, China. E-mail: jxhan{at}sdu.edu.cn

The rapid identification of bacteria in cerebrospinal fluid (CSF) is very important for patient management and antimicrobial therapies. We developed a 16S DNA microarray–based method that targets 16S rDNA and can directly detect bacteria from CSF without cultivation. Universal primers and specific probes were designed from the 16S rDNA sequence data retrieved directly from the GenBank database. The specificity of the assay is obtained through a combination of microarray hybridization and enzymatic labeling of the constructed specific probes. Cultivation-dependent assays were used as reference methods in the development and evaluation of the method. With the exception of Mycobacterium tuberculosis and Proteus mirabilis, forty-five positive blood culture media were successfully differentiated. When this procedure was applied directly to 100 CSF specimens, 29 specimens from 16 patients were positive by bacterial culture and 3 culture-positive CSF specimens produced no hybridized signals. The remaining 26 specimens were correctly identified, including one with mixed infection. The accuracy, sensitivity, and specificity of the assay can be increased further by designing more oligonucleotides for the microarray. This method is versatile and makes it possible to detect more bacteria in a single assay and discriminate different bacterial genera.

Key Words: 16S rDNA • microarray • cerebrospinal fluid • bacteria