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2016 ; 2016
(ä): 4210129
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Review of 16S and ITS Direct Sequencing Results for Clinical Specimens Submitted
to a Reference Laboratory
#MMPMID27366168
Payne M
; Azana R
; Hoang LM
Can J Infect Dis Med Microbiol
2016[]; 2016
(ä): 4210129
PMID27366168
show ga
We evaluated the performance of 16S and internal transcribed spacer (ITS) region
amplification and sequencing of rDNA from clinical specimens, for the respective
detection and identification of bacterial and fungal pathogens. Direct rDNA
amplification of 16S and ITS targets from clinical samples was performed over a
4-year period and reviewed. All specimens were from sterile sites and submitted
to a reference laboratory for evaluation. Results of 16S and ITS were compared to
histopathology, Gram and/or calcofluor stain microscopy results. A total of 277
16S tests were performed, with 64 (23%) positive for the presence of bacterial
DNA. Identification of an organism was more likely in microscopy positive 16S
samples 14/21 (67%), compared to 35/175 (20%) of microscopy negative samples. A
total of 110 ITS tests were performed, with 14 (13%) positive. The yield of
microscopy positive ITS samples, 9/44 (21%), was higher than microscopy negative
samples 3/50 (6%). Given these findings, 16S and ITS are valuable options for
culture negative specimens from sterile sites, particularly in the setting of
positive microscopy findings. Where microscopy results are negative, the limited
sensitivity of 16S and ITS in detecting and identifying an infectious agent needs
to be considered.