功能性便秘幼儿肠道菌群的16SrDNA和REP-PCR检测分析及效果评价

Detection analysis and effect evaluation on 16SrDNA and REP-PCR for intestinal flora in children with functional constipation

目的研究功能性便秘幼儿肠道菌群的16SrDNA和细菌基因组重复序列PCR技术(repetitive-element PCR,REP-PCR)检测分析及效果评价。方法选取我院2018年1~12月收治的功能性便秘患儿80例作为观察组,同期选取我院正常幼儿80例作为对照组。应用16SrDNA片段扩增和REP-PCR技术对两组幼儿的粪便进行检测分析,并对观察组患儿进行益生菌治疗。比较两组幼儿的肠道常驻菌及优势菌群在门和在属水平的差异性,计算出16SrDNA和REP-PCR检测的灵敏度和特异度。结果对照组肠道菌群中放线菌门水平明显低于观察组,对照组肠道菌群中变形菌门水平明显高于观察组(P<0.05),两组肠道菌群中无壁菌门、厚壁菌门、拟杆菌门、梭菌属门水平比较无明显差异(P<0.05);对照组肠道菌群中副杆状菌属、拟杆菌属水平明显低于观察组,对照组肠道菌群中普氏菌属水平明显高于观察组(P<0.05);16SrDNA检测的灵敏度为87.17%,特异度为92.03%,REP-PCR检测的灵敏度为85.29%,特异度为84.25%。结论应用16SrDNA和REP-PCR检测分析功能性便秘幼儿肠道菌群具有重要价值,患儿肠道菌群表现为放线菌门、副杆状菌属、拟杆菌属水平升高,变形菌门、普氏菌属水平降低,对治疗起到指导作用,值得在临床推广。

Objective To study the detection analysis and effect evaluation on 16 SrDNA and bacterial genome repetitive-element PCR(repetitive-element PCR, REP-PCR) for intestinal flora in children with functional constipation. Methods80 children patients with functional constipation who were admitted to our hospital from January to December 2018 were selected as the study subjects and were assigned to the observation group. In the same period, 80 healthy children in our hospital were selected as the control group. The fragment amplification of 16 SrDNA and REP-PCR techniques were used to detect and analyze the feces of the two groups of children, and the probiotics was used for treatment in the observation group. The differences were compared between the two groups of intestinal resident bacteria and dominant bacteria at the levels of phylum and genus. The sensitivity and specificity of 16 SrDNA and REP-PCR detection were calculated. Results The level of Actinobacteria in the intestinal flora in the control group was significantly lower than that in the observation group. The level of Proteobacteria in the intestinal flora in the control group was significantly higher than that in the observation group(P<0.05). There were no significant differences in the levels of Mycobacterium,Absidia, Bacteroides and Clostridium in the intestinal flora between the two groups(P<0.05);in the control group, the levels of Parabacteroides and Bacteroides in the intestinal flora were significantly lower than those in the observation group. The level of Plasmodium in the intestinal flora in the control group was significantly higher than that in the observation group(P<0.05);the sensitivity of 16 SrDNA was 87.17%, and the specificity was 92.03%. The sensitivity of REP-PCR was 85.29%, and the specificity was 84.25%. Conclusion It is of a great value to analyze the intestinal flora of children with functional constipation by 16 SrDNA and REP-PCR techniques. The intestinal flora of the children patients shows an increase in the levels of Actinobacteria, Parabacteroides, and Bacteroides, and an decrease in the levels of Proteobacteria and Plasmodium. It plays a guiding role in treatment and is worthy of clinical promotion.