[Rapid detection of Streptococcus pneumoniae and Haemophilus influenzae with DNA probes].
OBJECTIVE
To establish a method for rapid molecular detection of Streptococcus pneumoniae and Haemophilus influenzae in the early stage of infection.


METHODS
Specific DNA probes for Streptococcus pneumoniae and Haemophilus influenzae and 16 S rRNA universal probe of bacteria were synthesized by polymerase chain reaction (PCR) and labeled with biotin. The DNA of the bacteria, virus and fungi were hybridized with these probes respectively prior to application for examination of the clinical samples.


RESULTS
The DNA probes of 263, 351, and 370 bp were amplified by PCR. Streptococcus pneumoniae and Haemophilus influenzae reacted only with their corresponding probes, and no cross reaction of the bacterial universal probe with virus and fungi was noted. The method could detect bacterial DNA in as small amount as 1 ng. Of the 100 sputum specimens, 11 were found to be positive for Streptococcus pneumoniae and 8 for Haemophilus influenzae, with a positivity rate greater than that by sputum culture.


CONCLUSIONS
DNA probe detection is simple, rapid, and specific for clinical examination of Streptococcus pneumoniae and Haemophilus influenzae.
Fan
Hui-zhen
HZ
Department of Respiratory Diseases, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. huizhen2000@163.com
Yu
Hua-peng
HP
Huang
Wen-jie
WJ
D015342:DNA Probes; D006192:Haemophilus Infections / Q000382:microbiology; D006193:Haemophilus influenzae / Q000302:isolation & purification*; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D011008:Pneumococcal Infections / Q000382:microbiology; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013296:Streptococcus pneumoniae / Q000302:isolation & purification*
D015342:DNA Probes; D012336:RNA, Ribosomal, 16S