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Evaluating Children in the Ukraine for Colonization With the Intestinal Bacterium Oxalobacter formigenes, Using a
1Department of Pathology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, Florida, USA
Insights
A new DNA-based test quickly identifies Oxalobacter formigenes, a gut bacterium crucial for regulating oxalic acid. This method simplifies research into its role in health conditions like kidney stones and hyperoxaluria.
Area of Science:
- Microbiology
- Molecular Biology
- Human Health
Background:
- Oxalobacter formigenes is an anaerobic gut bacterium involved in oxalic acid homeostasis.
- Dysregulation of oxalic acid is linked to kidney stones, hyperoxaluria, and other health issues.
- Traditional methods for identifying O. formigenes are difficult and time-consuming.
Purpose of the Study:
- To develop a rapid, DNA-based method for identifying O. formigenes.
- To circumvent the limitations of culturing and isolation techniques.
- To facilitate research on the association between O. formigenes and human health.
Main Methods:
- Designed genus- and group-specific oligonucleotide sequences targeting the oxc gene.
- Utilized polymerase chain reaction (PCR) to amplify the 5' end of the oxc gene from genomic DNA.
- Validated the PCR-based detection system using 100 fecal cultures from children (0-12 years).
Main Results:
- The PCR assay successfully identified and subgrouped O. formigenes strains.
- Field testing demonstrated the ease and efficacy of the new detection system.
- Data revealed a colonization profile, suggesting O. formigenes colonization begins after crawling.
Conclusions:
- The developed PCR method allows for rapid identification and classification of O. formigenes.
- This advancement simplifies the study of O. formigenes' role in human health and disease.
- Future research on conditions like kidney stones and hyperoxaluria associated with O. formigenes absence is now more feasible.
Abstract:
Background: Oxalobacter formigenes is a recently discovered anaerobic bacterium residing in the gastrointestinal tracts of most vertebrates, including humans. Evidence suggests that this bacterium plays an important symbiotic relationship with its hosts by regulating oxalic acid homeostasis. Oxalic acid is a ubiquitous toxic by-product of metabolism associated with numerous pathologic conditions, including hyperoxaluia, cardiac myopathy and conductance disorders, kidney stones, and even death. Despite the potential importance of O. formigenes in several major health disorders, the difficulty in culturing, isolating, and identifying this fastidious anaerobe has limited research of its disease associations. Because O. formigenes must use two unique enzymes to catabolize oxalic acid, this bacterium appeared to be a suitable model for DNA-based identification, thereby circumventing the labor-intensive procedures currently used. Methods and Results: In this study, genus- and group-specific oligonucleotide sequences were designed corresponding to homologous regions residing in the oxc gene that enodes for oxalyl-coenzyme A decarboxylase. A polymerase chain reaction (PCR)-based amplification of the 5'end of this gene directly from genomic DNA isolated from various strains of O. formigenes was used to show that the genus- and group-specific oligonucleotide probes could identify and subgroup the bacterium. Field testing of this PCR-based detection system with 100 fecal cultures collected from children aged 0-12 years demonstrated the ease and efficacy with which O. formigenes can now be identified. Furthermore, these latter data provide a profile for the natural colonization of a human population with this intestinal bacterium. Conclusions: Development and use of this PCR-based detection system permit the rapid identification and classification of the gut-associated bacterium O. formigenes, thereby circumventing the need for the more labor-intensive and lengthy method currently used. The first field test of this detection system indicates that humans apparently do not become colonized with O. formigenes until they begin crawling about in the environment. Furthermore, studies investigating the association between several disorders (eg, kidney stones, irritable bowel syndrome, and hyperoxaluria) and the absence of the bacterium from the gut will now prove far easier.