Analysis On Intestinal Flora In Feeding Intolerance In Preterm Infants

Part Ⅰ: Analysis on Intestinal flora in feeding intolerance inpreterm infants by PCR-DGGE【Objective】PCR and Denaturing Gradient GelElectrophoresis(DGGE) were used to analyse the difference of intestinalflora in stool between the feeding intolerance group and feeding tolerancegroup (the control group).【Methods】Collect the stool samples from the first day (no feeding)after birth, feeding intolerance group showed clinical manifestations ofintolerance, feeding intolerance restored group return to nomal feedingbetween15feeding intolerance preterm infants and the control groupincluding15feeding tolerance preterm infants.Total bacteria DNA wasextracted from the stool samples.The V3regions of the16S rDNA of stoolbacteria were amplified.The characteristic profiles of DGGE were cut andrecycled by PCR,after sequence test,the bacterium type was determined.【Results】Compared with the control group, feeding intolerancegroup showed more than or less than one different DGGE band.theybelonged to Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis and Lactobacilliin using sequence test.【Conclusion】The diversity of the predominant bacteria between thefeeding intolerance group and the control group were different.Part Ⅱ： Quantification of intestinal flora in feedingintolerance in preterm infants by Real-Time PCR【Objective】 To observe the variation of Escherichia coli,Klebsiella pneumoniae, Enterococcus faecalis, Lactobacilliin andBifidobacteria in the feeding intolerance in preterm infants.【Methods】 The amount of Escherichia coli, Klebsiellapneumoniae, Enterococcus faecalis, Lactobacilliin and Bifidobacteria ofstool respectively from the first day (no feeding) after birth, feedingintolerance group showed clinical manifestations of intolerance, feedingintolerance restored group return to nomal feeding between15feedingintolerance preterm infants and the control group including15feedingtolerance preterm infants were measured by the Real-time quantitativePCR.【Results】 Logarithmic absolute value (lg copies/g) of Escherichiacoli from above three conditions were (2.62±0.22)、(5.47±1.28)、(3.04±0.70)in feeding intolerance respectively, and (2.56±0.19),(2.82±0.4),(2.80±0.39)in control group respectively. Logarithmic absolute value (lg copies/g) ofKlebsiella pneumoniae were (4.37±0.22),(6.56±0.27),(4.17±0.27) in feeding intolerance respectively, and (4.35±0.22),(4.19±0.14),(4.15±0.25)in control group respectively. Absolute value of Enterococcus faecaliswere (79.17±93.46),(42.84±47.57),(101.68±43.78) in feeding intolerancegroup respectively, and (70.16±78.41),(740.05±657.71),(104.57±38.39) incontrol group respectively. Absolute value of Lactobacilliin were204.03±25.57,326.04±82.42,677.73±559.49in feeding intolerance grouprespectively, and205.48±16.65、678.79±124.93、663.33±491.57in controlgroup respectively. Logarithmic absolute value (lg copies/g) ofBifidobacteria were4.79±0.07,5.27±0.17,5.65±0.25in feeding intolerancerespectively, and4.76±0.07,5.57±0.09,5.64±0.15in control grouprespectively. The above values had statistical difference between the feedingintolerance group and the control group when feeding intolerance occurring(P＜0.05).【Conclusion】 The number of Escherichia coli and Klebsiellapneumoniae was significantly higher if preterm infants appear feedingintolerance, which may be involved in the genesis of feeding intolerance.Enterococcus faecalis, Lactobacilliin and Bifidobacteria may play aprotective role.