Identification of Streptococcus species and Haemophilus influenzae by direct sequencing of PCR products from 16S-23SrDNA intergenic spacer regions
Lu, X.; Yang, C.; Li, L.; Yang, H.
Chinese Medical Journal 115(9): 1415-1417
2002
ISSN/ISBN: 0366-6999 PMID: 12411126 Document Number: 542717
Objective: To set up a rapid and simple method for identificating bacteria by 16S-23SrDNA intergenic spacer regions (ISRs). Methods: Polymorphic products of PCR from ISRs were selected on agarose gel and sequenced directly using purified fragments by excising the gel without cloning. Nucleotide sequences were compared with GenBank databases and analyzed by DNAMAN program. Results: There was only a single product in streptococcus genus after PCR amplification of 16S-23SrDNA ISRs. Five streptococcal species were obtained from 7 strains of streptococcus. Two major amplicons were consistently generated for 8 strains of Haemophilus influenzae (H. influenzae). The sequence data showed that they all belonged to H. influenzae type b on GenBank databases. Conclusion: PCR and direct sequencing of 16S-23SrDNA ISRs were very successful methods for bacterial species identification.