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Collection and Extraction of Saliva DNA for Next Generation Sequencing
Published on: August 27, 2014
[Proinflammatory cytokine content in the saliva of children suffering from chronic tonsillitis]
N V Boiko1, I V Stagnieva1, A S Kim1
1ENT Department of Rostov State Medical University, Rostov-on-Don, Russia, 344022.
Insights
Saliva analysis revealed elevated Interleukin-6 (IL-6) in children with chronic tonsillitis (CT). A diagnostic model combining IL-6 and Tumor Necrosis Factor-alpha (TNF-α) shows high accuracy for CT detection.
Area of Science:
- Pediatric immunology
- Otorhinolaryngology
- Biomarker discovery
Background:
- Chronic tonsillitis (CT) affects children, with potential links to inflammatory cytokines.
- Tumor Necrosis Factor-alpha (TNF-α) and Interleukins (IL-1β, IL-6) are key inflammatory mediators.
- Salivary cytokine levels in CT patients require further investigation.
Purpose of the Study:
- To investigate salivary cytokine profiles in children with chronic tonsillitis.
- To identify potential salivary biomarkers for CT diagnosis.
- To develop a mathematical model for CT diagnosis using cytokine levels.
Main Methods:
- Enzyme-linked immunosorbent assay (ELISA) was used to measure cytokine levels (TNF-α, IL-1β, IL-6) in saliva.
- Saliva samples were collected from 92 children with CT and 17 healthy controls.
- Statistical analysis, including Mann-Whitney assessment and logistic regression, was performed.
Main Results:
- Significantly higher levels of Interleukin-6 (IL-6) were observed in children with CT compared to healthy controls (median 12.5 vs 6.72, p=0.01).
- A diagnostic model based on TNF-α and IL-6 demonstrated high sensitivity (80.4%) and specificity (82.4%) for CT.
- IL-6 dominance in saliva is linked to the persistent antigenic challenge in toxic allergic CT.
Conclusions:
- Elevated salivary IL-6 is a significant indicator in children with chronic tonsillitis.
- A mathematical model combining TNF-α and IL-6 shows promise as an additional diagnostic tool for CT.
- Measuring salivary IL-6 is recommended as a supplementary diagnostic criterion for CT in children.
Introduction:
We have examined 92 children aged between 6 and 15, suffering from chronic tonsillitis (CT). Tumor necrosis factor (TNF-α) and interleukin 1β and 6 (IL-1β and IL-6) contents have been defined in saliva. The control set comprised 17 healthy children. Cytokine content was defined with the enzyme multiplied immunoassay sets (Vektor Best Ltd., Russia) by enzyme-linked immunosorbent assay. The statistic analysis and data processing were carried out with statistic analysis programs (version 3.2, R Foundation for Statistical Computing, Vienna, Austria).
Results:
The content of cytokines TNF-α, IL-1, IL-6 in CT children's saliva was high against the healthy children, yet the statistically significant differences were only noted for IL-6. In the CT group the median value of this factor (12.5) was significantly higher than in the control set (6.72) (p=0.01 in Mann-Whitney assessment). IL-6 was chosen as the basic factor for the mathematic model; its combinations in the form of a multi-factor logistic regression were given consideration. From out of the three possible models there was just one that had all the coefficients statistically significantly different from zero (TNF-α - IL-6). It was chosen as the basic diagnostic model for chronic tonsillitis. The created model's sensitivity is 80.4%, while its specificity is 82.4%.
Discussion:
The revealed IL-6 dominance in saliva at CT can be is attributable to permanent antigenic challenge characteristic of the toxic allergic CT since, as previously shown, there are living proliferating microorganisms in the palatal tonsil tissues and their blood- and lymph vessels at CT.
Conclusion:
The conducted ROC-analysis has demonstrated high sensitivity and specificity of the mathematical model, which enabled us to recommend determination of IL-6 in the saliva of the children suffering from CT as an additional diagnostic criterion.
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