Detecting The Variation Of Intestinal Microflora In Children With Rotavirus Enteritis By 16SrRNA Fluoresecent Quantificative PCR

Objectives To study the variation of intestinal microflora in children with rotavirus enteritis and compare with the healthy children in same age .To analyze the relationship between rotavirus infection and changes of intestinal microflora from microecology.To provide basis for the prevention and treatment of rotavirus enteritis with microecologic products. To understand practicality and feasibility of fluoresecent quantificative PCR in the field in order to generalize the application of the technique.Methods The faces of the children with rotavirus enteritis and the control was collected and the bacteria DNA was extracted. The primes of bifidobacteria, lactobacillus and escherichia coli with the 16SrRNA sequence of bacteria was designed, the qualitative of the bacteria through general PCR was obtained. The standard curve of fluorescent quantificative PCR was made by using a series of dilution of accurate bacteria DNA. The unknown samples were measured by PCR arid compared with the standard curve.Results Compared with the healthy children, the number of bifidobacteria and lactobacillus appeared to decrease in patients (P<0.05) ,however the number of Escherichia coli have no significant variation (P>0.05). The range of bacteria by fluorescent quantificative PCR was nearly similar with the result of the cultivation techniques from other researchers.