Immunocytochemical Method for Early Laboratory Diagnosis of Tuberculous Meningitis

A simple immunocytochemical method was standardized for the direct demonstration of mycobacterial antigen in cerebrospinal fluid (CSF) specimens of patients with tuberculous meningitis (TBM). CSF-cytospin smears were prepared from 22 patients with a clinical diagnosis of TBM and also from an equal number of patients with nontuberculous neurological diseases (disease control). Immunocytological demonstration of mycobacterial antigens in the cytoplasm of monocytoid cells was attempted, by using rabbit immunoglobulin G to Mycobacterium tuberculosis as the primary antibody. Of the 22 CSF-cytospin smears from TBM patients, 16 showed positive immunostaining, while all of the CSF-cytospin smears from the disease control showed negative immunostaining for mycobacterial antigen. The technical aspects of this immunocytological method for the demonstration of mycobacterial antigens are simple, rapid, and reproducible, as well as specific, and therefore can be applied for the early diagnosis of TBM, particularly in patients in whom bacteriological methods did not demonstrate the presence of M. tuberculosis in the CSF.     

改良聚合酶链反应快速诊断结核性脑膜炎

目的：建立改良的聚合酶链反应的实验方法，快速诊断结核性脑膜炎。方法：应用二步控温洽酶链反应技术检测脑脊液结核杆菌微量ＤＮＡ。结果：经是 铲的４０例结核性脑膜炎聚合酶链反应扩增脑脊液ＤＮＡ全部阳性，２４例不明原因的中枢神经系统感染患者，１６例为阳性，而其它病原体引起的中枢神经系统感染和正常对照组均无扩增反应。结论：二步控温聚合酶链反应技术快速诊断结核性脑膜炎与传统的检验方法相比较，具有快速、敏感、高     

高原牧区结核性脑膜炎4例病例诊断分析

目的 了解高原牧区结核性脑膜炎发病情况.方法对我院2005年11月～2010年9月住院的部分结核性脑膜炎患者43例进行了发病诊断治疗分析.结果初发病例37例,占发病86.1%;急危病例17例,占39.54%;单纯结核性脑膜炎者12例,占发病27.91%;肺结核合并结核性脑膜炎者22例,占发病51.16%,其他结核合并结核性脑膜炎者4例;结核性脑膜炎合并气管炎者5例.治愈2例,占4.65%;好转36例,占83.72%;总有效率88.37%.民族上牧区藏族牧民及儿童41例,占发病95.34%;年龄:20～39岁年龄段30例,占发病69.77%,最小2个月,最大69岁.结论高原牧区牧民结核性脑膜炎发病可能处在高发期,在当前结核病防治工作中要及时发现有效治疗高原牧区藏族牧民结核性脑膜炎病人.     

Evaluation of GeneXpert MTB/RIF for Diagnosis of Tuberculous Meningitis

Tuberculous meningitis (TBM) is the most severe form of tuberculosis. Microbiological confirmation is rare, and treatment is often delayed, increasing mortality and morbidity. The GeneXpert MTB/RIF test was evaluated in a large cohort of patients with suspected tuberculous meningitis. Three hundred seventy-nine patients presenting with suspected tuberculous meningitis to the Hospital for Tropical Diseases, Ho Chi Minh City, Vietnam, between 17 April 2011 and 31 December 2012 were included in the study. Cerebrospinal fluid samples were tested by Ziehl-Neelsen smear, mycobacterial growth indicator tube (MGIT) culture, and Xpert MTB/RIF. Rifampin (RIF) resistance results by Xpert were confirmed by an MTBDR-Plus line probe assay and all positive cultures were tested by phenotypic MGIT drug susceptibility testing. Overall, 182/379 included patients (48.0%) were diagnosed with tuberculous meningitis. Sensitivities of Xpert, smear, and MGIT culture among patients diagnosed with TBM were 59.3% (108/182 [95% confidence interval {CI}, 51.8 to 66.5%]), 78.6% (143/182 [95% CI, 71.9 to 84.3%]) and 66.5% (121/182 [95% CI, 59.1 to 73.3%]), respectively. There was one false-positive Xpert MTB/RIF test (99.5% specificity). Four cases of RIF resistance (4/109; 3.7%) were identified by Xpert, of which 3 were confirmed to be multidrug-resistant (MDR) TBM and one was culture negative. Xpert MTB/RIF is a rapid and specific test for the diagnosis of tuberculous meningitis. The addition of a vortexing step to sample processing increased sensitivity for confirmed TBM by 20% (P = 0.04). Meticulous examination of a smear from a large volume of cerebrospinal fluid (CSF) remains the most sensitive technique but is not practical in most laboratories. The Xpert MTB/RIF represents a significant advance in the early diagnosis of this devastating condition.     

Serum and Cerebrospinal Fluid Neuron-Specific Enolase for Diagnosis of Tuberculous Meningitis

Purpose

Late diagnosis and treatment lead to high mortality and poor prognosis in tuberculous meningitis (TbM). A rapid and accurate diagnosis is necessary for a good prognosis. Neuron-specific enolase (NSE) has been investigated as a biochemical marker of nervous tissue damage. In the present study, the usefulness of NSE was evaluated, and a cut-off value for the differential diagnosis of TbM was proposed.

Materials and Methods

Patient charts were reviewed for levels of serum and cerebrospinal fluid (CSF) NSE, obtained from a diagnostic CSF study of samples in age- and gender-matched TbM (n=15), aseptic meningitis (n=28) and control (n=37) patients.

Results

CSF/serum NSE ratio was higher in the TbM group than those of the control and aseptic groups (p=0.001). In binary logistic regression, CSF white blood cell count and CSF/serum NSE ratio were significant factors for diagnosis of TbM. When the cut-off value of the CSF/serum NSE ratio was 1.21, the sensitivity was 86.7% and the specificity was 75.4%.

Conclusion

The CSF/serum NSE ratio could be a useful parameter for the early diagnosis of TbM. In addition, the authors of the present study suggest a cut-off value of 1.21 for CSF/serum NSE ratio.     

Use of Roche AMPLICOR Mycobacterium tuberculosis PCR in Early Diagnosis of Tuberculous Meningitis

Several nucleic acid-based amplification tests are available for the detection of Mycobacterium tuberculosis, but few data are available on their use in the diagnosis of tuberculous meningitis (TBM). We performed a prospective study to assess the Roche AMPLICOR Mycobacterium tuberculosis PCR test (TB AMPLICOR) for use in the diagnosis of TBM and compared it with direct Ziehl-Neelsen staining of smears, radiometric culture for M. tuberculosis, and clinical and cerebrospinal fluid (CSF) findings. Eighty-three CSF specimens collected from 69 patients with suspected meningitis in South Africa were tested by TB AMPLICOR. On the basis of clinical and laboratory findings, 40 of these patients were treated for TBM and 29 patients were not treated for TBM. Ten CSF samples from 10 patients were positive by TB AMPLICOR. Seven of these 10 patients were classified as having definite TBM, 2 were classified as having probable TBM, and 1 was classified as having possible TBM. The sensitivity of TB AMPLICOR for detecting cases of definite and probable TBM in patients from whom CSF specimens had been collected less than 10 days into antituberculosis treatment was 60.0%. Specimens from all 29 patients not treated for TBM were negative by the TB AMPLICOR, giving a 100% specificity. TB AMPLICOR is therefore more sensitive than the combination of Ziehl-Neelsen staining of smears and radiometric culture for M. tuberculosis and is a rapid and highly specific diagnostic test for TBM.In the past few years there has been a global increase in the incidence of tuberculosis (TB), with an increase in the number of notifications of TB in England and Wales (7), other European countries (20), and the United States (2). Tuberculous meningitis (TBM) is rare in developed countries, but Mycobacterium tuberculosis is an important cause of meningitis in many developing countries, where facilities to confirm it as the cause of meningitis are least available. Without treatment, death occurs in virtually all patients with TBM, while delay in treatment results in a considerable risk of irreversible neurological damage. Therefore, rapid diagnosis is important but difficult, since the spectrum of disease is wide and abnormalities of the cerebrospinal fluid (CSF), although usually present, are very variable. Direct smears of CSF for acid-fast bacilli (AFB), although virtually diagnostic, are usually positive in fewer than 10% of cases of TBM (11, 26), while culture for M. tuberculosis takes up to 8 weeks and is also often negative. A clinical response to antituberculosis treatment is usual, but patients with TBM may deteriorate on appropriate treatment. Hence, a rapid and sensitive test is required for the diagnosis of TBM.A number of nucleic acid-based amplification tests, most of them based on PCR, have been developed for the detection of M. tuberculosis in clinical specimens. Although considerable data are now available on their use with respiratory specimens in the diagnosis of pulmonary tuberculosis, little is known of their role in diagnosing TBM from specimens of CSF. Only six published studies (in the English-language literature) of M. tuberculosis PCRs for the diagnosis of TBM have included more than 20 patients with suspected or confirmed TBM (Table ​(Table1).1). Those studies used primers targeting either the MPB 64 protein (15, 22, 23) or the insertion sequence IS6110 (12, 16, 19). Overall sensitivities for the diagnosis of TBM ranged from 33% (19) to 90.5% (15), with specificities ranging from 88.2% (23) to 100% (15, 19). From these data it is clear that the role of PCR in the diagnosis of TBM is undefined.

TABLE 1

Studies of TB PCR for the diagnosis of TBM

Comparison of Amplicor and GeneXpert MTB/RIF Tests for Diagnosis of Tuberculous Meningitis

There are no data about the comparative accuracy of commercially available nucleic acid amplification tests (GeneXpert MTB/RIF and Roche Amplicor) for the diagnosis of tuberculous meningitis (TBM). A total of 148 patients with suspected TBM were evaluated, and cultures served as the reference standard. The sensitivities and specificities (95% confidence interval [CI]) for the Amplicor and Xpert MTB/RIF tests were similar: 46 (31–60) versus 50 (33–67) and 99 (93–100) and 94 (84–99), respectively.     

Tuberculous Meningitis: Immune Reactions within the Central Nervous System

Cerebrospinal fluid (CSF) lymphocytes from two patients with tuberculous meningitis proliferated stronger than the corresponding peripheral blood lymphocytes (PBL) when stimulated with tuberculin purified protein derivative (PPD) in the lymphocyte transformation test after 3 days of culture. This might indicate an accumulation of specifically primed lymphocytes within the central nervous system. CSF lymphocytes and PBL from nine of ten patients with acute aseptic meningitis investigated as controls showed no or low responses when stimulated with. PPD, whereas the remaining patient displayed a significant proliferation of CSF lymphocytes, which was more pronounced than that of PBL. Stimulation with the mitogens phytohaemagglutinin, concanavalin A, and pokeweed mitogen gave lower proliferation of CSF lymphocytes compared with PBL in tuberculous and aseptic meningitis. Evaluation of the proliferative response of CSF lymphocytes compared with PBL on stimulation with PPD might be a useful complement in the diagnosis of tuberculous meningitis.     

利奈唑胺联合莫西沙星治疗结核性脑膜炎的效果

目的 探讨利奈唑胺联合莫西沙星治疗结核性脑膜炎的效果。方法 选取2017年5月~2018年6月我院收治的结核性脑膜炎患者64例,采用随机数字表法分为对照组和观察组,每组32例。对照组给予莫西沙星治疗,观察组在对照组基础上加用利奈唑胺,比较两组治疗前后腺苷脱氨酶水平、蛋白质、葡萄糖、白细胞计数及脑脊液压力,临床疗效及不良反应总发生率。结果治疗后,观察组蛋白质、葡萄糖、白细胞计数、脑脊液压力、腺苷脱氨酶水平优于对照组（P＜0.05）。观察组总有效率为93.75%,高于对照组81.25%（P＜0.05）。观察组不良反应总发生率为21.88%,高于对照组的18.75%（P＞0.05）。结论 利奈唑胺联合莫西沙星治疗结核性脑膜炎疗效显著,可有效改善脑脊液生化指标,降低脑脊液压力及腺苷脱氨酶水平,且不增加不良反应发生几率。     

Mycobacterial Dormancy Regulon Protein Rv2623 as a Novel Biomarker for the Diagnosis of Latent and Active Tuberculous Meningitis

The present study was designed to investigate Rv2623 antigen, a major dormancy regulon protein of Mycobacterium tuberculosis (MTB) in CSF of suspected latent and active tuberculous meningitis (TBM) patients. A total of 100 CSF samples from TBM (n = 31), suspected latent TBM (n = 22), and suitable noninfectious control subjects (n = 47) were collected and evaluated for Rv2623 antigen level using ELISA protocol. A significantly high (P < 0.05) mean absorbance was observed in samples of suspected latent TBM and active TBM patients as compared to non-TBM control patients. However, no significant difference in Rv2623 level was observed between suspected latent TBM and TBM patients. Our preliminary findings suggest that Rv2623 may be useful as a potential biomarker for the diagnosis of the latent as well as active TBM infection. Futher evaluation of this biomarker in large number of samples is therefore needed to confirm the result.     

结核性脑膜炎患者脑脊液基质金属蛋白酶-9的检测意义

在脑膜炎的发展过程中,基质金属蛋白酶-9(MMP-9)对降解基底膜和损伤血脑屏障发挥重要作用,结核分枝杆菌能够刺激单核-巨噬细胞过度表达MMP-9.为检测结核性脑膜炎患者脑脊液MMP-9水平,评价其对结核性脑膜炎诊断及治疗的意义,应用ELISA方法检测12例结核性脑膜炎和9例病毒性脑膜炎患者脑脊液MMP-9水平,并选取5例非肿瘤非感染性头痛患者脑脊液作对照组.结果显示,结核性脑膜炎脑脊液MMP-9的水平显著增高,中枢神经系统并发症发生率与MMP-9的水平相关.结论为脑脊液MMP-9水平对结核性脑膜炎的诊断和预后有参考价值.     

结核性腹膜炎诊断技术的研究进展

结核性腹膜炎在现代临床上属于重要结核疾病,因其临床表现缺乏特异性,以致目前仍为临床中较难诊断的一种结核疾病,更对结核性腹膜炎的及时有效治疗造成较严重困难,因此近几年结核性腹膜炎诊断技术一直是结核病诊治的攻克课题。现笔者在本文中对结核性腹膜炎诊断技术的研发应用进展进行统一综述,归纳各项结核性腹膜炎诊断技术的优点与欠缺之处。     

降钙素原在病毒性脑膜炎和细菌性脑膜炎鉴别诊断中的应用

目的探讨降钙素原（PCT）在病毒性脑膜炎和细菌性脑膜炎鉴别诊断中的应用价值。方法通过对27例细菌性脑膜炎和32例病毒性脑膜炎患者血清PCT值的检测和比较,从而分析降钙素原在其鉴别诊断中的临床意义。结果细菌性脑膜炎患者血中的PCT平均浓度为37.84 ng/ml,病毒性脑膜炎患者的PCT平均浓度为0.41 ng/ml,两者相比差异有统计学意义（χ2=28.45,P〈0.01）,特异性分别为100%和96.9%。结论降钙素原对病毒性脑膜炎和细菌性脑膜炎的鉴别诊断具有临床应用价值。