Pathogen Identification in Suspected Cases of Pyogenic Spondylodiscitis

Pyogenic spinal infection continues to represent a worldwide problem. In approximately one-third of patients with pyogenic spondylodiscitis, the infectious agent is never identified. Of the cases that lead to organismal identification, bacteria are more commonly isolated from the spine rather than f...

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Main Authors: Ahmad Farajzadeh Sheikh, Azar D. Khosravi, Hamed Goodarzi, Roohangiz Nashibi, Alireaza Teimouri, Azim Motamedfar, Reza Ranjbar, Sara Afzalzadeh, Mehrandokht Cyrus, Mohammad Hashemzadeh
Format: Article
Language:English
Published: Frontiers Media S.A. 2017-03-01
Series:Frontiers in Cellular and Infection Microbiology
Subjects:
Online Access:http://journal.frontiersin.org/article/10.3389/fcimb.2017.00060/full
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author Ahmad Farajzadeh Sheikh
Ahmad Farajzadeh Sheikh
Azar D. Khosravi
Azar D. Khosravi
Hamed Goodarzi
Hamed Goodarzi
Hamed Goodarzi
Roohangiz Nashibi
Roohangiz Nashibi
Alireaza Teimouri
Alireaza Teimouri
Azim Motamedfar
Reza Ranjbar
Sara Afzalzadeh
Mehrandokht Cyrus
Mohammad Hashemzadeh
Mohammad Hashemzadeh
author_facet Ahmad Farajzadeh Sheikh
Ahmad Farajzadeh Sheikh
Azar D. Khosravi
Azar D. Khosravi
Hamed Goodarzi
Hamed Goodarzi
Hamed Goodarzi
Roohangiz Nashibi
Roohangiz Nashibi
Alireaza Teimouri
Alireaza Teimouri
Azim Motamedfar
Reza Ranjbar
Sara Afzalzadeh
Mehrandokht Cyrus
Mohammad Hashemzadeh
Mohammad Hashemzadeh
author_sort Ahmad Farajzadeh Sheikh
collection DOAJ
description Pyogenic spinal infection continues to represent a worldwide problem. In approximately one-third of patients with pyogenic spondylodiscitis, the infectious agent is never identified. Of the cases that lead to organismal identification, bacteria are more commonly isolated from the spine rather than fungi and parasites. This study applied universal prokaryotic 16S rRNA PCR as a rapid diagnostic tool for the detection of bacterial agents in specimens from patients suspected of pyogenic spondylodiscitis. Gram and Ziehl-Neelsen staining were used as a preliminary screening measure for microbiologic evaluation of patient samples. PCR amplification targeting 16S rRNA gene was performed on DNA extracted from 57 cases including specimens from epidural abscesses, vertebral, and disc biopsies. Positive samples were directly sequenced. MRI findings demonstrated that disc destruction and inflammation were the major imaging features of suspected pyogenic spondylodiscitis cases, as 44 cases showed such features. The most common site of infection was the lumbar spine (66.7%), followed by thoracic spine (19%), the sacroiliac joint (9.5%), and lumbar-thoracic spine (4.8%) regions. A total of 21 samples amplified the 16S rRNA-PCR product. Sanger sequencing of the PCR products identified the following bacteriological agents: Mycobacterium tuberculosis (n = 9; 42.9%), Staphylococcus aureus (n = 6; 28.5%), Mycobacterium abscessus (n = 5; 23.8%), and Mycobacterium chelonae (n = 1; 4.8%). 36 samples displayed no visible 16S rRNA PCR signal, which suggested that non-bacterial infectious agents (e.g., fungi) or non-infectious processes (e.g., inflammatory, or neoplastic) may be responsible for some of these cases. The L3–L4 site (23.8%) was the most frequent site of infection. Single disc/vertebral infection were observed in 9 patients (42.85%), while 12 patients (57.15%) had 2 infected adjacent vertebrae. Elevated erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) inflammatory markers were noted in majority of the patients. In conclusion, microbiological methods and MRI findings are vital components for the proper diagnosis of pyogenic spondylodiscitis. Our findings suggest that molecular methods such as clinical application of 16S rRNA PCR and sequencing may be useful as adjunctive diagnostic tools for pyogenic spondylodiscitis. The rapid turnaround time of 16S rRNA PCR and sequencing submission and results can potentially decrease the time to diagnosis and improve the therapeutic management and outcome of these infections. Although S. aureus and M. tuberculosis were the most common causes of pyogenic spinal infections in this study, other infectious agents and non-infectious etiologies should be considered. Based on study results, we advise that antibiotic therapy should be initiated after a definitive etiological diagnosis.
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spelling doaj.art-26cd13e3823a41e6a50e1371e52bee722022-12-22T03:16:35ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882017-03-01710.3389/fcimb.2017.00060218573Pathogen Identification in Suspected Cases of Pyogenic SpondylodiscitisAhmad Farajzadeh Sheikh0Ahmad Farajzadeh Sheikh1Azar D. Khosravi2Azar D. Khosravi3Hamed Goodarzi4Hamed Goodarzi5Hamed Goodarzi6Roohangiz Nashibi7Roohangiz Nashibi8Alireaza Teimouri9Alireaza Teimouri10Azim Motamedfar11Reza Ranjbar12Sara Afzalzadeh13Mehrandokht Cyrus14Mohammad Hashemzadeh15Mohammad Hashemzadeh16Infectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranDepartment of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranInfectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranDepartment of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranInfectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranDepartment of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranMolecular Biology Research Center, Baqiyatallah University of Medical ScienceTehran, IranDepartment of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranDepartment of Infectious Diseases, Razi Teaching Hospital, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranDepartment of Neurosurgery, Golestan Teaching Hospital, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranDepartment of Neurosurgery, Tehran Medical Science Branch, Islamic Azad UniversityTehran, IranDepartment of Radiology, Razi Teaching Hospital, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranMolecular Biology Research Center, Baqiyatallah University of Medical ScienceTehran, IranDepartment of Infectious Diseases, Razi Teaching Hospital, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranDepartment of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranInfectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranDepartment of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical SciencesAhvaz, IranPyogenic spinal infection continues to represent a worldwide problem. In approximately one-third of patients with pyogenic spondylodiscitis, the infectious agent is never identified. Of the cases that lead to organismal identification, bacteria are more commonly isolated from the spine rather than fungi and parasites. This study applied universal prokaryotic 16S rRNA PCR as a rapid diagnostic tool for the detection of bacterial agents in specimens from patients suspected of pyogenic spondylodiscitis. Gram and Ziehl-Neelsen staining were used as a preliminary screening measure for microbiologic evaluation of patient samples. PCR amplification targeting 16S rRNA gene was performed on DNA extracted from 57 cases including specimens from epidural abscesses, vertebral, and disc biopsies. Positive samples were directly sequenced. MRI findings demonstrated that disc destruction and inflammation were the major imaging features of suspected pyogenic spondylodiscitis cases, as 44 cases showed such features. The most common site of infection was the lumbar spine (66.7%), followed by thoracic spine (19%), the sacroiliac joint (9.5%), and lumbar-thoracic spine (4.8%) regions. A total of 21 samples amplified the 16S rRNA-PCR product. Sanger sequencing of the PCR products identified the following bacteriological agents: Mycobacterium tuberculosis (n = 9; 42.9%), Staphylococcus aureus (n = 6; 28.5%), Mycobacterium abscessus (n = 5; 23.8%), and Mycobacterium chelonae (n = 1; 4.8%). 36 samples displayed no visible 16S rRNA PCR signal, which suggested that non-bacterial infectious agents (e.g., fungi) or non-infectious processes (e.g., inflammatory, or neoplastic) may be responsible for some of these cases. The L3–L4 site (23.8%) was the most frequent site of infection. Single disc/vertebral infection were observed in 9 patients (42.85%), while 12 patients (57.15%) had 2 infected adjacent vertebrae. Elevated erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) inflammatory markers were noted in majority of the patients. In conclusion, microbiological methods and MRI findings are vital components for the proper diagnosis of pyogenic spondylodiscitis. Our findings suggest that molecular methods such as clinical application of 16S rRNA PCR and sequencing may be useful as adjunctive diagnostic tools for pyogenic spondylodiscitis. The rapid turnaround time of 16S rRNA PCR and sequencing submission and results can potentially decrease the time to diagnosis and improve the therapeutic management and outcome of these infections. Although S. aureus and M. tuberculosis were the most common causes of pyogenic spinal infections in this study, other infectious agents and non-infectious etiologies should be considered. Based on study results, we advise that antibiotic therapy should be initiated after a definitive etiological diagnosis.http://journal.frontiersin.org/article/10.3389/fcimb.2017.00060/fullMRIpyogenic spondylodiscitisbiopsypolymerase chain reactionbacterial agents
spellingShingle Ahmad Farajzadeh Sheikh
Ahmad Farajzadeh Sheikh
Azar D. Khosravi
Azar D. Khosravi
Hamed Goodarzi
Hamed Goodarzi
Hamed Goodarzi
Roohangiz Nashibi
Roohangiz Nashibi
Alireaza Teimouri
Alireaza Teimouri
Azim Motamedfar
Reza Ranjbar
Sara Afzalzadeh
Mehrandokht Cyrus
Mohammad Hashemzadeh
Mohammad Hashemzadeh
Pathogen Identification in Suspected Cases of Pyogenic Spondylodiscitis
Frontiers in Cellular and Infection Microbiology
MRI
pyogenic spondylodiscitis
biopsy
polymerase chain reaction
bacterial agents
title Pathogen Identification in Suspected Cases of Pyogenic Spondylodiscitis
title_full Pathogen Identification in Suspected Cases of Pyogenic Spondylodiscitis
title_fullStr Pathogen Identification in Suspected Cases of Pyogenic Spondylodiscitis
title_full_unstemmed Pathogen Identification in Suspected Cases of Pyogenic Spondylodiscitis
title_short Pathogen Identification in Suspected Cases of Pyogenic Spondylodiscitis
title_sort pathogen identification in suspected cases of pyogenic spondylodiscitis
topic MRI
pyogenic spondylodiscitis
biopsy
polymerase chain reaction
bacterial agents
url http://journal.frontiersin.org/article/10.3389/fcimb.2017.00060/full
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