Sheikh Ahmad Farajzadeh, Khosravi Azar D, Goodarzi Hamed, Nashibi Roohangiz, Teimouri Alireaza, Motamedfar Azim, Ranjbar Reza, Afzalzadeh Sara, Cyrus Mehrandokht, Hashemzadeh Mohammad
Infectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical SciencesAhvaz, Iran; Department of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical SciencesAhvaz, Iran.
Infectious and Tropical Diseases Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical SciencesAhvaz, Iran; Department of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical SciencesAhvaz, Iran; Molecular Biology Research Center, Baqiyatallah University of Medical ScienceTehran, Iran.
Front Cell Infect Microbiol. 2017 Mar 9;7:60. doi: 10.3389/fcimb.2017.00060. eCollection 2017.
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 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 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 -PCR product. Sanger sequencing of the PCR products identified the following bacteriological agents: ( = 9; 42.9%), ( = 6; 28.5%), ( = 5; 23.8%), and ( = 1; 4.8%). 36 samples displayed no visible 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 PCR and sequencing may be useful as adjunctive diagnostic tools for pyogenic spondylodiscitis. The rapid turnaround time of PCR and sequencing submission and results can potentially decrease the time to diagnosis and improve the therapeutic management and outcome of these infections. Although and 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.
化脓性脊柱感染仍然是一个全球性问题。在大约三分之一的化脓性脊椎间盘炎患者中,感染源从未被确定。在那些能够确定病原体的病例中,从脊柱中分离出细菌的情况比真菌和寄生虫更为常见。本研究应用通用原核生物PCR作为一种快速诊断工具,用于检测疑似化脓性脊椎间盘炎患者标本中的细菌病原体。革兰氏染色和齐-尼氏染色被用作患者样本微生物学评估的初步筛查措施。对从57例病例中提取的DNA进行靶向基因的PCR扩增,这些病例包括硬膜外脓肿、椎体和椎间盘活检标本。阳性样本直接进行测序。MRI结果表明,椎间盘破坏和炎症是疑似化脓性脊椎间盘炎病例的主要影像学特征,44例病例显示了此类特征。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%)和胸腰椎(4.8%)区域。共有21个样本扩增出PCR产物。PCR产物的桑格测序鉴定出以下细菌病原体:金黄色葡萄球菌(n = 9;42.9%)、表皮葡萄球菌(n = 6;28.5%)、大肠埃希菌(n = 5;23.8%)和肺炎克雷伯菌(n = 1;4.8%)。36个样本未显示可见的PCR信号,这表明非细菌感染源(如真菌)或非感染性过程(如炎症或肿瘤)可能是其中一些病例的病因。L3-L4部位(23.8%)是最常见的感染部位。9例患者(42.85%)观察到单个椎间盘/椎体感染,而12例患者(57.15%)有2个相邻椎体感染。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椎间盘炎的重要组成部分。我们的研究结果表明,诸如PCR临床应用和测序等分子方法可能作为化脓性脊椎间盘炎辅助诊断工具有用。PCR和测序提交及结果的快速周转时间可能会减少诊断时间,并改善这些感染的治疗管理和结果。尽管金黄色葡萄球菌和表皮葡萄球菌是本研究中化脓性脊柱感染的最常见原因,但应考虑其他感染源和非感染性病因。基于研究结果,我们建议在明确病因诊断后开始抗生素治疗。