Usefulness Of16S RDNA Sequencing In The Detection And Identification Of Pathogens In PD Peritonitis

Background:Peritoneal dialysis (PD) has been an established treatment modality for end-stage renal disease (ESRD) for several years. Peritonitis remains a serious complication of peritoneal dialysis and an important cause of morbidity and mortality in PD patients. Traditional culture methods are time-consuming、expensive and, can be false negative, particularly after antibiotic therapy. Among molecular techniques,16S rDNA Sequencing can provide a sensitive method for identifying infectious organisms, including slow-growing and fastidious bacteria.Bacterial16S rDNA contain hypervariable regions and conserved regions. By using universal primers or specific probes,we can get specific target sequences for bacterial species identification.Aims:To apply16S rDNA sequencing to identify common bacterial pathogens directly from the dialysates.Methods:To evaluate the usefulness of16S rDNA sequencing in the detection of Pathogens, we compared the molecular method with bacteriological culture for the analysis of dialysates from PD peritonitis patients.69samples from PD peritonitis patients and control samples from 18PD patients without peritonitis were examined by16S rDNA sequencing analysis and by conventional bacteriological culture.Results:Of the69samples collected from PD peritonitis patients,63.8%(44/69) were positive by culture,71%(49/69) were positive by direct sequencing analysis(P>0.05). In32of the44culture-positive samples, the same microorganisms were confirmed by sequencing analysis, and the others showed discrepant results as compared with culture study. In8of the25culture-negative samples, organisms were detected by the molecular method. Of the18PD peritonitis samples collected after antibiotic therapy,50%(9/18) were positive by culture,61.1%(11/18) were positive by sequencing analysis(P>0.05). In the control samples from patients without PD peritonitis, none microorganisms were found by direct sequencing or bacteriological culture.Conclusions:In our study, the molecular method based on the16S rDNA sequencing identified more etiological agents in comparison to culture method, without statistical difference. But this molecular method is time-saving, inexpensive, simple to operate and is not affected by antibiotics. To conclude,16S rDNA sequencing is a well established tool for rapid and accurate identification of bacterial pathogens and can complement culture methods in the diagnosis of PD peritonitis.