Determining the utility of standard hospital microbiology testing: Comparing standard microbiology cultures with DNA sequence analysis in patients with chronic sinusitis

Abstract

Objective To demonstrate DNA sequencing analysis (DNAsa) of sinus cultures in patients with CRS is a reliable method of detecting pathogens in polymicrobial CRS infections. Methods After obtaining Institutional Review Board approval for this prospective cohort study, we selected a random sample of 50 patients with CRS at Medstar Georgetown University Hospital between September 2016 and March 2017. We defined CRS as a history of rhinosinusitis refractory to maximal medical therapy and prior endoscopic sinus surgery. Patients demonstrating active purulence in a sinus cavity were prospectively selected to undergo standard hospital cultures (SHC) and DNAsa cultures. Organisms identified in both methods were compared for each patient. Results Specimens were obtained from 29 female and 16 male patients with a mean age of 50 years. A total of 45 cultures were included in our final analysis; five cultures were excluded after inappropriate laboratory processing. Results from these patients were compared and analyzed. Cohen's weighted kappa analysis showed agreement between the two testing methods in identifying predominant microorganisms. DNAsa detected 31.9% more microorganisms compared to SHC ( P < 0.05). When multiple microorganisms were detected, DNAsa yielded more positive results compared to SHC ( P < 0.05). Conclusions DNAsa detects all microorganisms identified by SHC as well as predominant microorganisms not detected by SHC. Thus molecular pathogen identification may be more reliable for identifying multiple microorganisms as compared to standard culture techniques that identify only one or two microorganisms. In recalcitrant cases of CRS, DNAsa may provide better guidance in selection of appropriate antimicrobial treatment.

Increased lipopolysaccharide content is positively correlated with glucocorticoid receptor‐beta expression in chronic rhinosinusitis with nasal polyps

“…Therefore, the simple method of nasal bacterial culture limits the understanding of the pathological mechanism of CRSwNP, 14 and molecular diagnosis of submucosal bacterial clones and histopathological observation are more sensitive and diagnostic than simple bacterial culture. 26,27 LPS is considered to be a trigger for inflammatory responses. LPS activates inflammatory signaling pathways mainly through the classic Toll-like receptor 4 (TLR4) receptor on the cell surface, stimulating the body to produce inflammatory factors, and leading to tissue remodeling.…”

“…This result may be an important reason for the inconsistency in the results. Therefore, the simple method of nasal bacterial culture limits the understanding of the pathological mechanism of CRSwNP, 14 and molecular diagnosis of submucosal bacterial clones and histopathological observation are more sensitive and diagnostic than simple bacterial culture 26,27 …”

Detection of Microbiota in Post Radiation Sinusitis

“…The 16S ribosomal RNA is highly conserved within bacterial and fungal species and can therefore be detected for microbial identification and quantification. 16 Pyrosequencing is a method of DNA sequencing in which inorganic pyrophosphate is released with the DNA polymerase reaction and incorporation of each nucleotide. Pyrophosphate is converted to adenosine triphosphate (ATP) by sulfurylase, and a proportionate amount of light is produced by luciferase.…”

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“…However, whether it can be used for the differential diagnosis of odontogenic maxillary sinusitis from maxillary sinus needs further study [22][23][24][25][26][27][28], whereas cytokines and chemokines are currently detected in local tissues, and detection in serum is relatively rare. Rapoport et al tried to compare microbial DNA probes with a conventional microbial culture to find the pathogenic microbial as well as chemosensitivity and found that DNA probes are more accurate and reliable than traditional methods [29]. However, the detection efficiency is limited and needs further expansion.…”