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Updated: Jun 16, 2026

Oral Biofilm Sampling for Microbiome Analysis in Healthy Children
Published on: December 31, 2017
Test, Treat, Repopulate™ method reduces red complex bacteria and stabilizes the oral microbiome
Rachelle Beattie McAliley1, Lora Hooper2, Kami Hoss3
1ProBiora Health, LLC, Sarasota, FL, United States.
Abstract:
Despite advances in dental hygiene and clinical interventions, preventable conditions such as tooth decay and gum disease remain widespread public health concerns. Recent research implicates complex, polymicrobial interactions as central drivers of disease pathogenesis, which often limits the effectiveness of traditional therapeutic approaches. Here, researchers evaluate the effectiveness of the Test, Treat, Repopulate method, a multi-phase oral health protocol combining scaling and root planing, targeted antibiotic therapy, use of a pH-balanced, prebiotic containing toothpaste and rinse, and daily oral probiotic use on the health of the oral microbiome. Retrospective analysis of RT-qPCR salivary diagnostic data from 38 de-identified U.S. patients, collected before and after treatment during routine care, demonstrated significant and sustained reductions in key oral pathogens following the combined treatment approach. Seven bacterial species were significantly reduced post-treatment, including Treponema denticola, Tannerella forsythia, Prevotella intermedia, Campylobacter rectus, Fusobacterium nucleatum, Fusobacterium nucleatum subsp. animalis, and Streptococcus mutans. Additional pathogens, including Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis, were also reduced. The protocol had minimal impact on the fungal species Candida albicans, highlighting an area for potential optimization. Additionally, four viral targets were assessed pre- and post-protocol, making this one of the first studies to evaluate a dental intervention's effects on oral viruses. Comparison of the cohort with 35,570 pre-treatment salivary diagnostic tests across the United States shows that baseline pathogen distributions reflect national patterns, supporting the generalizability of the protocol. A structured protocol for patients with elevated red complex bacteria is detailed, providing clinicians with a data-driven, reproducible framework for precision oral healthcare. These findings underscore the potential for microbiome-focused interventions to achieve measurable, lasting improvements in oral health across diverse populations.
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