Pathogen Identification in Suspected Cases of Pyogenic Spondylodiscitis.
Pathogen Identification in Suspected Cases of Pyogenic Spondylodiscitis.
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DOI：
10.3389/fcimb.2017.00060
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发表时间：
2017
影响因子：
5.7
通讯作者：
Hashemzadeh M
中科院分区：
文献类型：
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作者：
Sheikh AF;Khosravi AD;Goodarzi H;Nashibi R;Teimouri A;Motamedfar A;Ranjbar R;Afzalzadeh S;Cyrus M;Hashemzadeh M
化脓性脊柱感染仍然是一个世界性的问题。在大约三分之一的化脓性脊柱炎患者中，感染源从未被确定。在导致生物体鉴定的情况下，细菌通常是从脊柱中分离出来的，而不是真菌和寄生虫。本研究应用通用原核16S rRNA PCR作为快速诊断工具，检测疑似化脓性脊柱炎患者标本中的细菌病原体。革兰氏染色和齐尔-尼尔森染色作为初步筛选措施，对患者样本进行微生物学评估。对57例硬膜外脓肿、椎和椎间盘活检标本提取的DNA进行16S rRNA基因PCR扩增。阳性样品直接测序。MRI结果显示，椎间盘破坏和炎症是疑似化脓性脊柱炎病例的主要影像学特征，44例患者表现出上述特征。最常见的感染部位是腰椎（66.7%），其次是胸椎（19%）、骶髂关节（9.5%）和腰胸椎（4.8%）区域。共有21份样本扩增了16S rRNA-PCR产物。PCR产物Sanger测序鉴定出以下细菌因子：结核分枝杆菌（n = 9, 42.9%）、金黄色葡萄球菌（n = 6, 28.5%）、脓肿分枝杆菌（n = 5, 23.8%）和龟分枝杆菌（n = 1, 4.8%）。36个样本未显示可见的16S rRNA PCR信号，这表明非细菌感染因子（如真菌）或非感染过程（如炎症或肿瘤）可能是其中一些病例的原因。l3 ~ l4部位是最常见的感染部位（23.8%）。单椎间盘/椎体感染9例（42.85%），相邻两椎体感染12例（57.15%）。大多数患者的红细胞沉降率（ESR）和c反应蛋白（CRP）炎症标志物升高。总之，微生物学方法和MRI结果是正确诊断化脓性脊柱炎的重要组成部分。我们的研究结果表明，分子方法如16S rRNA PCR和测序的临床应用可能有助于作为化脓性脊柱炎的辅助诊断工具。16S rRNA PCR的快速周转时间以及测序提交和结果可以潜在地减少诊断时间，改善这些感染的治疗管理和结果。虽然金黄色葡萄球菌和结核分枝杆菌是本研究中最常见的化脓性脊柱感染原因，但也应考虑其他感染性病原体和非感染性病因。根据研究结果，我们建议抗生素治疗应在明确的病因诊断后开始。
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.