Broad-range amplification and sequencing of the rpoB gene: a novel assay for bacterial identification in clinical microbiology.

ABSTRACT: The rpoB gene has been proposed as a promising phylogenetic marker for bacterial identification, providing theoretically improved species-level resolution compared to the 16S rRNA gene for a range of clinically important taxa. However, its utility in diagnostic microbiology has been limited by the lack of broad-range primers allowing for its amplification from most species with a single PCR assay. Here, we present an assay for broad-range partial amplification and Sanger sequencing of the rpoB gene. To reduce cross-reactivity and allow for rpoB amplification directly from patient samples, primers were based on the dual priming oligonucleotide principle. The resulting amplicon is ~550 base pairs in length and appropriate for species-level identification. Systematic i

SUBMITTER: Bivand JM 

PROVIDER: S-EPMC11324016 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature