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Rapid detection of the novel human pathogen Pantoea piersonii: advancements in methodology
Louise O'Connor1, Elizabeth Minogue1, Shaunagh Carolan1
1Molecular Diagnostics Research Group, School of Biological and Chemical Sciences University of Galway, Ireland.
Abstract:
Pantoea piersonii a novel bacterium isolated from the International Space Station (ISS) presents a unique challenge for microbial monitoring in spaceflight and more recently in clinical environments. Identification of the organism currently involves culture, followed by whole genome sequencing and analysis of generated sequences. Since the MALDI-TOF profile of this pathogen is absent from the database and 16S rRNA sequencing fails to resolve its identity to the nearest neighbour, a definitive genetic marker is required for unambiguous identification of the organism. Given the increase in the number of reported clinical cases, there exists a need for a rapid method for identification of the organism which could be utilised in a range of environments including the clinical setting. This study describes the design, development and validation of a specific and sensitive real-time PCR assay for the specific detection of P. piersonii. The assay targets a unique region of the malate dehydrogenase gene, confirmed through comparative genomic analysis. We demonstrate the performance of the assay in terms of analytical specificity, sensitivity, and robustness, ensuring its suitability for both space microbiology applications and clinical use.
Insights
A new real-time PCR assay specifically detects Pantoea piersonii, a bacterium found on the International Space Station and in clinical settings. This rapid method aids microbial monitoring in space and healthcare.
Area of Science:
- Microbiology
- Molecular Biology
- Space Science
Background:
- Pantoea piersonii, a novel bacterium, was isolated from the International Space Station (ISS).
- Current identification methods (culture, whole genome sequencing, MALDI-TOF, 16S rRNA sequencing) are insufficient for rapid and unambiguous identification.
- P. piersonii poses challenges for microbial monitoring in spaceflight and clinical environments due to its increasing clinical relevance.
Purpose of the Study:
- To develop and validate a specific and sensitive real-time PCR assay for the rapid detection of P. piersonii.
- To provide a definitive genetic marker for unambiguous identification of the organism.
- To create a method suitable for diverse environments, including clinical settings.
Main Methods:
- Comparative genomic analysis to identify a unique genetic marker.
- Design and development of a real-time PCR assay targeting a specific region of the malate dehydrogenase gene.
- Validation of the assay's analytical specificity, sensitivity, and robustness.
Main Results:
- A specific and sensitive real-time PCR assay for P. piersonii was successfully designed, developed, and validated.
- The assay targets a unique region within the malate dehydrogenase gene, ensuring high specificity.
- Demonstrated performance metrics confirm the assay's suitability for its intended applications.
Conclusions:
- The developed real-time PCR assay provides a rapid and reliable method for identifying P. piersonii.
- This assay addresses the need for improved microbial detection in both spaceflight and clinical settings.
- The validated assay enhances the ability to monitor and manage P. piersonii in diverse environments.
