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Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Analysis of the Relationship between Gut Flora Levels in Childhood Obese Population and Normal Healthy Population
Yaoqing Feng1, Xia Si1, Ruifang Zhu1
1School of Nursing, Shanxi Medical University, 56 Xinjian South Road, Taiyuan, Shanxi Province 030001, China.
Aims:
To explore the study of the relationship between the level of gut flora in childhood obese people and normal healthy people based on the analysis of machine learning.
Materials And Methods:
The stools of 54 normal weight, 53 overweight, and 59 obese children from May 2021 to May 2022 were selected. And DNA was extracted, and primers specific for the four bacteria were designed according to the specificity of the four bacteria to the 16 S rDNA gene sequences of the bacteria to be tested, and real-time fluorescence quantitative PCR reactions were performed to compare whether there was any difference in the number of the four bacteria between the three groups. Results. The results of agarose gel electrophoresis showed that the PCR amplification products of all four target bacteria showed clear bands at the corresponding positions, and no nonspecific bands appeared. When compared with the marker, the size matched with the target fragment, indicating good primer specificity. The comparison between normal body recombinant, super recombinant, and obese groups was statistically significant (P < 0.05) for rectal eubacteria, polymorphic anaplasma, bifidobacteria spp., and lactobacilli. The median number of bifidobacteria in the three groups was significantly higher than the median number of rectal eubacteria, polymorphomycetes, and lactobacilli. The difference in comparison was statistically significant (P < 0.05). Stratified analysis of children's age revealed that normal body composition of Lactobacillus decreased with increasing age, and the difference was statistically significant (P < 0.05).
Conclusion:
An increase in rectal eubacteria and a decrease in polymorphomycetes, bifidobacteria spp., and lactobacilli may be associated with the development of obesity. The numbers of rectal eubacteria, polymorphic methanobacteria, bifidobacteria spp., and lactobacilli in the intestine of normal weight and obese children were less affected by sex and age.
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