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| Main Authors: | , , , , , , , , , , , , |
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| Format: | Artículo científico |
| Language: | en |
| Published: |
PeerJ
2026
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| Subjects: | |
| Online Access: | https://pubmed.ncbi.nlm.nih.gov/41943823/ |
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Table of Contents:
- Comparison of primers for the pathogenicity factors and in environmental isolates from the Texas Coastal Bend region of the Gulf of Mexico. Sama, Vinoshna Planas-Costas, Githzette Buck, Gregory W Peasari, John Reddy Mott, Joanna Kaiser, Kyra Grace Ramirez, Gabriel D Kostoch, Brian Galvan, Alexis Crow, Bruce Vasquez, Emilio Lee, Lauren Shevaun Idowu, Mary Boluwatife Vibrio vulnificus Sigma Factor Bacterial Proteins Gulf of America Polymerase Chain Reaction Texas DNA Primers Virulence Factors Vibrio Infections Humans Longitudinal Studies The gram-negative marine bacterium may cause necrotizing fasciitis in persons with predisposing conditions such as type II diabetes and immunosuppression. Inhabitants of the Texas Coastal Bend have a high prevalence of such conditions and are at risk from contracting vibriosis upon flooding of marine and estuarine waters during storms, leading to increased risk of morbidity and mortality. While digital droplet polymerase chain reaction (ddPCR) and other methods may quickly identify pathogenic bacteria with more sensitivity and specificity, the time to optimize the assay, cost and dependence on uninterrupted power supplies make this method less useful in locations with minimal resources. This investigation undertook a longitudinal study from 2009, 2014, and 2021 to determine if primers for the cytolysin-hemolysin gene, , and the alternative sigma factor, , could be consistently used in polymerase chain reaction (PCR) assays to identify strains taken from local waters. Inconsistencies in expected results led to the hypothesis that inability to reproduce PCR assays resulted from technical concerns or the locus exhibited instability or genome plasticity. Whole genome sequences of ATCC 27562 were retrieved on a high-performance cluster through NCBI Primer Blast and used to design additional and primers. PCR detecting a 265 bp amplicon of was performed for 12,410 sequences to determine specificity and sensitivity, producing amplicons from 17 of 32 strains (53%) but not any other species within the genus , demonstrating sensitivity. No members of other gram-negative or gram-positive bacteria yielded amplicons, indicating specificity. New primers were designed to produce 393 bp amplicons. Due to loss of strains from hurricane outages, the number of Coastal Bend isolates studied decreased from 2009 to 2021, but 21 of 22 strains (96%) showed amplicons and 22 of 23 strains (96%) revealed amplicons. Primers for were analyzed within versions of the sequence for single nucleotide variants (SNVs). Two SNVs were seen in the area bound by the forward primer and four within the annealing region of the reverse primer, but SNV analysis reveals that these mutations may affect primer binding; however, further research is warranted. Limitations of the study were small sample sizes. These findings support the hypothesis that technical issues, rather than inherent genetic instability of the locus, caused the lack of PCR reproducibility, and confirm as a target for detecting pathogenic in coastal waters of the Texas Coastal Bend region. Future goals will be to analyze larger numbers of south Texas isolates to allow monitoring for infections that may occur from post-hurricane floods resulting from climate change.