Establishment of PCR sequence-based typing for identifying bacteria based on 16S rDNA in the contaminate blood products

Abstract

Objective To establish a PCR sequence-based typing(PCR-SBT)method for identifying the bacteria accurately and rapidly.Methods The full-length of 16S rDNA was amplified with a set of universal primers.The PCR products were purified and sequenced directly.The related sequences were obtained from the GenBank database using BLAST search software and the species of the bacteria were identified according to the multiple sequence alignment and homology comparison of the 16S rDNA sequences by CLUSTAL X software.Results The results showed that it has been established a PCR-SBT method for analysis 16S rDNA nucleotide sequences by multi-primer pairs and 13 standard bacteria strains were obtained about 1400 bp of the 16S rDNA sequences.The sequences of these bacteria were 100%concordance with the known standard nucleotide sequences,which confirmed the accuracy of the method.The difierent unknown strains that isolated from platelet and cord blood in the laboratory were successfully identified using the 16S rDNA PCR-SBT methed.Estimation of the minimal levels of the genomic DNA detection Was 0.2 ng in the PCR reaction.Conclusion It suggested that the PCR-SBT based on 16S rDNA was reliable and could identify the bacteria rapidly and accuracy.It can detect the bacterial contamination of blood products as early as possible and has a potential application value in the direct treatment of transfusion related bacterial contamination. Key words: Bacterial contamination of blood; 16S rDNA; PCR sequence-based typing