Design and validation of Dolosigranulum pigrum specific PCR primers using the bacterial core genome

Sci Rep. 2023 Apr 14;13(1):6110. doi: 10.1038/s41598-023-32709-y.

Abstract

Dolosigranulum pigrum-a lactic acid bacterium that is increasingly recognized as an important member of the nasal microbiome. Currently, there are limited rapid and low-cost options for confirming D. pigrum isolates and detecting D. pigrum in clinical specimens. Here we describe the design and validation of a novel PCR assay targeting D. pigrum that is both sensitive and specific. We designed a PCR assay targeting murJ, a single-copy core species gene identified through the analysis of 21 D. pigrum whole genome sequences. The assay achieved 100% sensitivity and 100% specificity against D. pigrum and diverse bacterial isolates and an overall 91.1% sensitivity and 100% specificity using nasal swabs, detecting D. pigrum at a threshold of 1.0 × 104 D. pigrum 16S rRNA gene copies per swab. This assay adds a reliable and rapid D. pigrum detection tool to the microbiome researcher toolkit investigating the role of generalist and specialist bacteria in the nasal environment.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Primers
  • DNA, Bacterial
  • Gram-Positive Bacterial Infections* / microbiology
  • Gram-Positive Cocci*
  • Humans
  • Polymerase Chain Reaction
  • RNA, Ribosomal, 16S / genetics

Substances

  • RNA, Ribosomal, 16S
  • DNA Primers
  • DNA, Bacterial

Supplementary concepts

  • Dolosigranulum pigrum