Keywords
PCR
dermatoses in children
Abstract
Infectious dermatoses are one of the most common pathologies of the skin and subcutaneous fat among the pediatric population. Therefore, there is a need to study the contribution of the skin microbiome to the development of skin diseases, as well as to search for informative approaches for the routine detection of microorganisms (MO) associated with infectious dermatoses. Real-time polymerase chain reaction (RT-PCR) is a promising method for routine detection of opportunistic MO in skin microbiome samples. RT-PCR, unlike next-generation sequencing (NGS), is well adapted for practical healthcare and allows quantitative microbial analysis of skin microbiome samples. In addition, RT-PCR is devoid of the disadvantages of bacterial culture-based methods.
Objective. To assess the capabilities of the RT-PCR method in the detection of opportunistic microorganisms from the skin microbiome samples.
Materials and methods. The study included 13 children consulting ambulatory dermatologists with diagnosed skin rashes. Microbiota analysis was performed with RT-PCR by using the reagent kits “BacScreen” and “MycoseScreen” (DNA-Technology, LLC, Russia). Statistical analysis of data was carried out using R‑4.4.0 (release 2024-04-24).
Results. Bacterial DNA was detected in 92.3 % of the samples, while fungal DNA — in 80.8 %. For samples from the affected skin areas, we found a higher number of opportunistic MO and a greater total level of bacterial DNA, with a more frequent detection rate of bacteria belonging to the genera Staphylococcus and Acinetobacter, bacteria of the order Enterobacterales, and Candida parapsilosis. The highest level of bacterial DNA was observed in samples from the affected skin areas of patients with atopic dermatitis (107.5–108.2 GE/sample), which in average 10,000 times exceeded the amount of bacterial DNA for other samples.
Conclusion. RT-PCR detected bacterial and fungal DNA in most samples from the surface of the skin, usually in levels ranging 103 to 105 GE/sample.
For citation
Savcheno NV, Kornilov DO, Simarzina VM, Tryapitsyn MA, Nechaeva DM, Bekhter AA, et al. Pilot study on real-time PCR capabilities for the detection of opportunistic microorganisms in skin microbiome samples. USMU Medical Bulletin. 2024;(2):61–74. (In Russ.). EDN: https://elibrary.ru/RXECGY.
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Copyright © 2024 Savchenko N. V., Kornilov D. O., Simarzina V. M., Tryapitsyn M. A., Nechaeva D. M., Bekhter A. A., Itani T. M., Zornikov D. L., Voroshilina E. S.