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Updated: Jan 7, 2026
Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Changes in the Gut Microbiome is Influenced by the Level of Control and Treatment in Asthma
Ewelina Harcęko-Zielińska1, Aleksandra Górska1, Jan Romantowski1
1Department of Allergology, Medical University of Gdańsk, Gdańsk, Poland.
Background:
The influence of intestinal microorganisms on the development and course of allergic diseases has recently been the subject of intensive research, but studies describing changes in the intestinal microbiome of asthma patients in response to altering factors are still scarce.
Objective:
(1) the analysis of eating habits composition of intestinal microbiota and BMI in asthma patients compared to the control group, (2) the comparison of the results of the analyzed parameters in asthma patients and in the control group, (3) the analysis of asthma treatment results depending on the composition of intestinal microbiota.
Methods:
Clinical stool isolates were cultured and genetic material was sequenced. The study included 49 subjects with asthma and a control group of 18 healthy volunteers. Clinical data was collected through questionnaires on the most frequently reported symptoms and the FFQ questionnaire. The composition of intestinal microbiota was determined using the traditional breeding method (the serial dilution method was used) followed by 16S rRNA sequencing.
Results:
Patients with asthma reported the greatest severity of clinical symptoms in all the body systems examined.The most common cause of the aberrant stool test results was E. coli, with titers <106. The was no difference in the dietary habits between the asthma patients and the control group. Alpha and beta diversity, was significantly lower in asthma patients compared to the control group. Asthma patients had lower abundance of Faecalibacterium vs healthy volunteers. Statistically significant depletion of Oscilospirales, Anaerovoracaceea, was demonstrated in patients with uncontrolled asthma compared to controlled and partially controlled asthma. In patients taking glucocorticoids (oral and inhaled) enriched intestinal microbiota in Anaerovoracaceae and Christensenellaceae and depleted Faecalibacterium were observed.
Conclusion:
Patients with asthma showed less richness and diversity in the composition of their intestinal microbiota compared to the control group.
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