Development of a DNA microarray assay for rapid detection of fifteen bacterial pathogens in pneumonia

BMC Microbiology

article

Ma, Xiuqing¹  Li, Yanqin¹  Liang, Yuan¹  Liu, Yang¹  Yu, Ling¹  Li, Chunsun¹  Liu, Qiqi²  Chen, Liangan¹ 

[1] Department of Pulmonary & Critical Care Medicine, Chinese PLA General Hospital;Department of Biotechnology, Beijing Institute of Radiation Medicine

关键词: DNA microarray;    Bacteria;    Pneumonia;   

DOI  :  10.1186/s12866-020-01842-3

学科分类：放射科、核医学、医学影像

来源: BioMed Central

【 摘 要 】

The rapid identification of pathogenic bacteria is important for determining an appropriate antimicrobial therapy for pneumonia, but traditional bacterial culture is time-consuming and labourious. The aim of this study was to develop and evaluate a DNA microarray assay for the simultaneous detection of fifteen bacterial species directly from respiratory tract specimens in patients with pneumonia. These species included Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Mycoplasma pneumoniae, Enterococcus faecalis, Enterococcus faecium, Enterobacter cloacae, Stenotrophomonas maltophilia, Burkholderia cepacia, Legionella pneumophila and Chlamydia pneumoniae. The 16S rDNA genes and other specific genes of each pathogen were chosen as the amplification targets, amplified via multiplex polymerase chain reaction (PCR), and hybridized to oligonucleotide probes in a microarray. The DNA microarray detection limit was 103 copies/μL. Nineteen standard strains and 119 clinical isolates were correctly detected with our microarray, and 3 nontarget species from 4 clinical isolates were not detected. Additionally, bacterial pathogens were accurately identified when two or three bacterial targets were mixed together. Furthermore, the results for 99.4% (156/157) of clinical specimens were the same as those from a conventional assay. We developed a DNA microarray that could simultaneously detect various bacterial pathogens in pneumonia. The method described here has the potential to provide considerable labour and time savings due to its ability to screen for 15 bacterial pathogens simultaneously.

【 授权许可】

CC BY|CC0