Application of next generation sequencing technology in the detection of pathogenic bacteria in synovial fluid of prosthetic joint infection / 中华骨科杂志

ABSTRACT

Objective To investigate the role of next generation sequencing technology in the detection of pathogenic bacteria in synovial fluid of prosthetic joint infection.Methods Nine samples of synovial fluid specimens of prosthetic joint infection patients with positive microbial culture from October,1 2016 to April 1,2017 were collected.Each specimen (200 μl) was used for next generation sequencing.Total DNA was extracted from synovial fluid samples.The collected DNA samples were amplified by PCR in the V4 region of 16S rDNA gene.The amplified products were sequenced using the Illumina Miseq platform,2× 250 bp double-end sequencing strategy.The sequencing results were compared with the SILVA database to analyze the types of bacteria and relative abundance in the DNA samples.A total of 200 μl sterile double-distilled deionized water was used as control.Results Nine cases of microbial culture positive prosthetic joint infection synovial fluid DNA samples were sequenced by 16S rDNA amplicon sequencing and yielded 3 132 415 high-quality reads and 3 752 operational taxonomic units (OTU).At the level of bacteria,a total of 9 different bacterial gates were detected on 9 DNA samples.At the level of bacteria,34 different bacteria were detected by 16S rDNA amplicon sequencing.Each DNA sample was detected by 16S rDNA amplicon sequencing and the bacterial genus was identical to that of laboratory culture.16S rDNA amplicon sequencing detected more species of bacteria [6(3,9.5)] than bacterial cultures [(1.0(1.0,1.0)].There was statistically significant difference in the number of bacteria detected in the same specimen between the 16S rDNA amplicon sequencing and the laboratory culture (Z=2.533,P=0.011).Among them,the dominant bacterial population (highest abundance) detected by 16S rDNA amplicon sequencing in four DNA samples was consistent with the results of laboratory culture.Conclusion In the prosthetic joint infection,the 16S rDNA amplicon sequencing technology can accurately detect pathogens that are consistent with the laboratory culture,and can detect other bacteria outside the laboratory culture.This technology can provide the basis for clinical diagnosis and antibiotic selection.