학술논문
Design and validation of Dolosigranulum pigrum specific PCR primers using the bacterial core genome.
Document Type
Article
Author
Source
Subject
*BACTERIAL genomes
*WHOLE genome sequencing
*LACTIC acid bacteria
*SENSITIVITY & specificity (Statistics)
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Language
ISSN
2045-2322
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 × 104D. 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. [ABSTRACT FROM AUTHOR]