Spinal brucellosis: a review

Brucellosis is a zoonosis of worldwide distribution, relatively frequent in Mediterranean countries and in the Middle East. It is a systemic infection, caused by facultative intra-cellular bacteria of the genus Brucella, that can involve many organs and tissues. The spine is the most common site of musculoskeletal involvement, followed by the sacroiliac joints. The aim of this study was to assess the clinical, biological and imaging features of spinal brucellosis.     

Clinical and MRI findings of brucellar spondylodiscitis

OBJECTIVE: The aim of this retrospective study was to report the clinical features and MR imaging findings of patients with brucellar spondylodiscitis. MATERIALS AND METHODS: Twenty-two patients with spondylodiscitis, recruited among 152 patients with brucellosis referred from the Department of Infectious Diseases. Patients were diagnosed based on positive clinical findings, > or =1/160 titers of brucella agglutination tests and/or positive blood cultures. Magnetic resonance imaging (MRI) was performed to all of the patients with spondylodiscitis. Signal changes and enhancement of vertebral bodies, involvement of paravertebral soft tissues and epidural spaces, nerve root and cord compression and abscess formation were assessed. RESULTS: All of the patients (n=22; 7 F, 15 M) had > or =1/160 titers of brucella agglutination test and blood culture was positive in 9. A great majority of the patients had involvement at only one vertebrae level (n=21, 95.5%), whereas one patient (4.5%) had multilevel involvement. In MRI, eight patients had soft tissue involvement and three had abscess formation. All cases had vertebral and discal enhancement. Additionally epidural extension was detected in four cases, posterior longitudinal ligament (PLL) elevation in five cases and root compression in two cases. CONCLUSION: Brucella is still a public health problem in endemic areas. MRI is a highly sensitive and non-invasive imaging technique which should be first choice of imaging in the early diagnosis of spondylodiscitis.     

绵羊附睾种布鲁氏菌019株外膜蛋白基因序列比较研究

目的证实新疆分离绵羊附睾种布鲁氏菌019株与参考菌株的差异性。方法采用PCR扩增绵羊附睾种布鲁氏菌019株外膜蛋白的Omp31、Omp25、Omp22、Omp2a、Omp2b基因,并进行序列比较分析。结果绵羊附睾种布鲁氏菌019株的5种外膜蛋白基因序列与绵羊种布鲁氏菌参考株在核苷酸水平和氨基酸水平上均存在差异。结论绵羊附睾种布鲁氏菌019株是独特的新疆地方株。     

布鲁氏菌bp26和OMP10基因的原核表达和鉴定

目的研究布鲁氏菌外膜蛋白OMP10和bp26基因的表达、克隆与测序,并对其进行初步的血清学鉴定。方法从布鲁氏菌基因组中获得OMP10和bp26基因连接入PMD18-T克隆质粒并测序,克隆入融合表达载体PGEX-4T-1,构建重组质粒PGEX-4T-1/OMP10和PGEX-4T-1/bp26,在大肠杆菌中将该蛋白表达。用western-blot分析重组表达的GST-OMP10和GST-bp26的免疫学特性。结果成功构建了PGEX-4T-1/OMP10和PGEX-4T-1/bp26原核表达载体,并在大肠杆菌中成功表达了OMP10和bp26基因,布鲁氏菌免疫动物血清能特异性识别所表达的蛋白。结论布鲁氏菌外膜蛋白OMP10和bp26基因的成功表达,它可以与布鲁氏菌免疫血清产生特异性结合反应,为之后的抗原性以及免疫原性研究打下良好的物质基础。     

布氏杆菌BCSP_(31)/pVAX1重组表达质粒的构建及其免疫保护效果的研究

目的构建布氏杆菌BCSP31/pVAX1重组表达质粒,免疫BALB/c小鼠,观察其免疫应答及免疫保护效果。方法克隆BCSP31基因,构建BCSP31/pVAX1重组表达质粒。转染COS7细胞,免疫组化检测其BCSP31蛋白抗原的表达。BCSP31/pVAX1重组表达质粒肌肉注射免疫BALB/c小鼠,观察特异性体液免疫和细胞免疫效果。进一步用1.25×104布氏杆菌A544强毒株腹腔攻毒,观察BCSP31/pVAX1重组表达质粒的免疫保护效果。结果扩增的BCSP31保护性抗原基因片断在牛、羊和猪布氏杆菌株中具有高度保守性,构建的BCSP31/pVAX1重组表达质粒在COS7细胞有目的蛋白抗原的表达。制备的BCSP31/pVAX1重组表达质粒肌肉免疫BALB/c小鼠4次,ELISA检测BCSP31/pVAX1重组表达质粒产生了较高水平的特异性IgG抗体,并随免疫次数增加抗体的滴度也递增,且抗体亚型以IgG2a为主,表明主要引起Th1型的免疫应答。FCM检测BCSP31/pVAX1重组表达质粒的CD4+/CD8+比值明显下降,说明激发了显著的CTL效应。MTT测定BCSP31/pVAX1重组表达质粒的淋巴细胞增殖能力与对照组比较差异显著。布氏杆菌A544强毒株攻毒后,动物存活及脾组织细菌数与对照组比较有显著性差异,说明BCSP31/pVAX1重组表达质粒能够产生有效的保护效果。结论研制的BCSP31/pVAX1重组表达质粒免疫BALB/c小鼠能产生高水平的特异性体液免疫和细胞免疫,以细胞免疫为主,并产生有效的免疫保护效果,为布氏杆菌病新型疫苗的研究奠定了基础。     

Interleukin-10 gene polymorphisms and susceptibility to brucellosis in Iranian patients

Background: Interleukin-10 (IL-10) is a Th2-type cytokine that inhibits macrophage activation. It is known that production of IL-10 is affected by its gene promoter polymorphisms. Objective: To investigate the relationship between IL-10 gene promoter polymorphisms and susceptibility to brucellosis. Methods: One hundred and ninety patients with brucellosis and 81 healthy animal husbandmen who owned infected animals and consumed their contaminated dairy products were included in this study. All individuals were genotyped for three bi-allelic IL-10 gene promoter polymorphisms at positions -1082(G/A), -819(T/C), and -592(A/C) using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The distribution of C alleles at positions -592 and -819 of IL-10 were significantly higher in patients than in the healthy animal husbandmen (p=0.034 and p=0.0086, respectively). IL-10 ATA single and double haplotypes were significantly higher in controls, compared to the patients (p= 0.0278 and p=0.013, respectively). Conclusion: According to the results higher frequency of C alleles at positions -592 and -819 of IL-10 in patients may be considered as genetic factors for susceptibility to brucellosis.     

绵羊种布鲁氏菌019株侵染绵羊巨噬细胞cDNA文库的构建

目的构建绵羊种布鲁氏菌019株侵染绵羊肺泡巨噬细胞的cDNA文库。培养绵羊肺泡巨噬细胞,用绵羊种布鲁氏菌019株侵染绵羊巨噬细胞,用试剂盒构建侵染后绵羊肺泡巨噬细胞的cDNA文库,并对文库中的部分克隆进行测序。结果表明,构建的绵羊种布鲁氏菌019株侵染绵羊巨噬细胞cDNA文库达到建库要求;所测序列与牛的基因组编码序列同源性最高。在绵羊种布鲁氏菌019株侵染绵羊肺泡巨噬细胞过程中,以50∶1(细菌∶细胞)的个数比,侵染3.5h,能有效构建侵染后绵羊肺泡巨噬细胞的cDNA文库。     

1株粗糙型布鲁氏菌诱导株RB71的生物学特性研究

目的 通过研究1株粗糙型布鲁氏菌诱导株RB71的毒力及免疫效果,进一步验证其生物学特性,探讨其作为疫苗株的潜力。方法 通过药敏片扩散法检测诱导株RB71对β-内酰胺类及非β-内酰胺类抗生素的耐受程度;通过对小鼠脾脏载菌量测定,评价诱导株RB71对感染小鼠的免疫保护率并确定其最小免疫剂量;利用小鼠感染模型进行毒力试验,比较诱导株RB71与亲本株的毒力差异。结果 诱导后的布鲁氏菌RB71株与亲本株相比,对β-内酰胺类抗生素和非β-内酰胺类抗生素的敏感性未发生改变;诱导株RB71对感染小鼠具有一定的保护效果,最小免疫剂量为10⁹ CFU/只;诱导株RB71与亲本株相比毒力更小,安全性更高。结论 本研究验证了布鲁氏菌诱导株RB71对抗生素的敏感性,其具有良好的免疫原性且具有较低的毒力,具备成为疫苗株的潜力。     

应用CRISPR/Cas13a快速鉴定布鲁氏菌

目的 建立一种基于CRISPR/Cas13a的布鲁氏菌鉴定方法.方法 以布鲁氏菌BCSP31基因的保守区为靶标区域设计特异的引物和crRNA,建立鉴定布鲁氏菌的CRISPR/Cas13a方法.通过对纯菌株以及临床需氧血培养阳性菌液检测评价该方法的特异性和敏感性;通过梯度稀释法评估其检测下限.结果 建立了基于CRISPR...     

脊柱布氏杆菌感染的临床特征表现及抗菌方案的研究

目的探讨脊柱布氏杆菌感染的临床特征表现,比较多西环素+利福平、多西环素+利福平+左氧氟沙星的抗菌效果。方法回顾性分析2017年1月至2019年1月我院收治的78例脊柱布氏杆菌感染病人,分析其临床特征表现,并根据抗菌方案的不同分为观察组和对照组,每组39例,对照组行多西环素+利福平治疗,观察组行多西环素+利福平+左氧氟沙星治疗,观察两组治疗效果、临床症状及不良反应发生情况,采用疼痛数字评定量表(numerical rating scale,NRS)对病人关节疼痛程度予以评估。结果78例病人均有不同程度脊柱活动受限,其中关节疼痛78例(100.00%)、发热78例(100.00%)、乏力25例(32.05%)、多汗34例(43.59%)、肌肉痛12例(15.38%);CT检查显示病椎及相邻椎体边缘有不同程度破坏及明显骨质增生,病椎间隙狭窄;MRI检查显示病变椎旁软组织异常T1WI低信号、T2WI高信号;布氏杆菌特异性血清凝集试验均呈阳性,血白细胞水平异常11例(14.10%),C反应蛋白水平异常15例(19.23%),红细胞沉降率增快7例(8.97%)。治疗6周后,观察组总有效率为94.87%,明显高于对照组(79.49%,P<0.05);观察组体温、NRS评分明显低于同期对照组(P<0.05);观察组不良反应发生率为15.38%,与对照组(10.26%)比较,差异无统计学意义(P<0.05)。结论脊柱布氏杆菌感染的临床特征表现多样,对临床诊断及治疗有指导作用,相比于多西环素+利福平抗菌方案,多西环素+利福平+左氧氟沙星抗菌方案治疗效果更为显著,且具有较高安全性,值得推广。