脑脊液早期分泌性靶抗原-6水平的动态变化及对结核性脑膜炎的诊断价值

目的探讨脑脊液早期分泌性靶抗原-6(ESAT-6)水平的动态变化对结核性脑膜炎早期诊断的意义。方法结脑病人50例,其中确诊者10例,临床诊断40例,非结核病对照40例,健康对照10例。间接酶联免疫吸附试验检测各组ESAT-6水平。结果间接ELISA方法检测脑脊液中的ESAT-6,敏感性88%,特异性92%,结脑组早期脑脊液ESAT-6含量较对照组明显升高,抗痨治疗6 w后脑脊液ESAT-6含量明显下降。结论应用间接酶联免疫吸附试验测定脑脊液中ESAT-6水平诊断结脑是可靠的,其动态变化有助于判断结脑预后。     

酶联免疫法检测脑脊液阿拉伯糖甘露糖脂抗体诊断结核性脑 …

目的 评估检测阿拉伯糖甘露糖脂抗体（ＬＡＭ－ＩｇＧ）对结核性脑膜炎的诊断价值。方法 应用酶联免疫吸附试验检测脑脊液中的ＬＡＭ－ＩｇＧ。结果 ７３例结核性脑膜炎ＬＡＭ－ＩｇＧ阳性率为５１％,３６例对照组均呈阴性结果,特异性为１００％,结论 检测脑脊液中ＬＡＭ－ＩｇＧ对诊断结核性脑膜炎有较高的应用价值。     

MTB分泌蛋白抗原85B-早期分泌靶抗原6融合蛋白对结核性胸膜炎的辅助诊断价值

目的 研究MTB分泌蛋白抗原85B(Ag85B)-早期分泌靶抗原6(ESAT6)融合蛋白在结核性胸膜炎辅助诊断中的价值。方法 研究对象为2014年1月至2015年1月于河北省胸科医院住院的结核性胸膜炎(结核组)患者45例,非结核性胸膜炎(非结核组)患者26例,以及体检健康人群(健康组)25名。应用流式细胞仪分别检测结核分枝杆菌Ag85B-ESAT6融合蛋白刺激前后结核组、非结核组外周血、胸腔积液及健康组外周血中γ干扰素(IFN-γ)的含量,采用免疫组织化学染色法检测结核组与非结核组患者组织(通过手术或穿刺活检获得的胸膜组织)中CD4 ⁺和CD8 ⁺细胞的表达水平。采用SPSS 13.0软件进行统计学处理,数据符合正态分布的组间比较采用t检验,以P<0.05为差异有统计学意义。 结果 MTB Ag85B-ESAT6融合蛋白刺激前后,结核组外周血中IFN-γ的含量分别为(42.63±10.51)pg/ml和(401.90±72.54)pg/ml;非结核组分别为(38.97±7.08)pg/ml和(40.04±6.80)pg/ml;健康组分别为(39.61±7.28)pg/ml和(39.86±6.97)pg/ml。结核组胸腔积液中IFN-γ的含量分别为(411.91±41.56)pg/ml和(1342.67±167.96)pg/ml;非结核组分别为(47.99±11.49)pg/ml和(48.76±11.25)pg/ml。刺激前结核组胸腔积液中IFN-γ的含量明显高于非结核组,两组比较差异有统计学意义(t=55.194,P=0.000);刺激后结核组外周血单个核细胞产生的IFN-γ含量相对于刺激前及刺激后的其他两组比较,差异均有统计学意义(t=32.879、33.211和33.204,P值均为0.000);刺激后结核组与刺激前及非结核组刺激前后胸腔积液中IFN-γ的含量比较,差异均具有统计学意义(t=36.085、51.478和51.499,P值均为0.000)。免疫组织化学染色结果显示结核组与非结核组患者胸膜组织中的CD4 ⁺和CD8 ⁺积累光密度值(结核组:16349.91±2376.36和10525.77±1164.86;非结核组:1853.64±670.40和1327.15±175.55)差异均具有统计学意义(t=14.381和19.127,P值均为0.000)。 结论 结核分枝杆菌Ag85B-ESAT6融合蛋白刺激外周血和胸腔积液后均明显提高结核特异性IFN-γ的含量,对结核性胸膜炎的辅助诊断有较高的临床应用价值。     

脑脊液腺苷脱氨酶检测对结核性脑膜炎的诊断价值

目的：探讨脑脊液腺苷脱氨酶（ ADA）在结核性脑膜炎诊断中的价值。方法对结核性脑膜炎患者治疗前后脑脊液ADA的含量进行比较和分析。结果结核性脑膜炎患者早期ADA含量增高,随病情好转ADA含量下降。结论脑脊液ADA含量检测对结核性脑膜炎的诊断、疗效观察、预后有重要的价值。     

脑脊液细胞学在结核性脑膜炎与隐球菌性脑膜炎鉴别诊断中的价值

目的探讨脑脊液细胞学(CSFc)在结核性脑膜炎(TBM)与隐球菌性脑膜炎(CM)鉴别诊断中的价值。方法选择西京医院2009—2013年门诊及住院部收治的CM患者30例(隐脑组)和TBM患者30例(结脑组),回顾性分析两组患者的临床表现、脑脊液生化指标及CSFc指标。结果两组患者颅内压及脑脊液、糖、蛋白质、氯化物水平比较,差异均无统计学意义(P0.05)。两组患者白细胞计数和淋巴细胞分数比较,差异无统计学意义(P0.05);隐脑组患者单核细胞分数高于结脑组,中性粒细胞分数低于结脑组(P0.05)。结论 CSFc在CM与TBM鉴别诊断中的价值有限,临床因积极进行病原学检测以减少漏诊、误诊。     

结核分枝杆菌早期分泌抗原靶-6（ESAT-6）对人THP-1单核细胞IL-12产生的影响

目的研究结核分枝杆菌早期分泌抗原靶-6（ESAT-6）对人THP-1单核细胞IL-12产生的影响及其作用的细胞信号传导机制。方法应用亲和层析方法纯化大肠埃希菌表达重组的结核分枝杆菌ESAT-6抗原,采用ELISA法检测不同浓度ESAT-6刺激人THP-1单核细胞IL-12产生的影响,以及对LPS刺激的反应,应用各种细胞信号传导通路抑制剂来探讨ESAT-6诱导单核细胞IL-12产生相关可能的信号通路。结果结核分枝杆菌ESAT-6分泌抗原在2.5～10g/ml浓度范围能依赖性地刺激人THP-1单核细胞产生IL-12（p70 及其亚单位p40）。细胞信号传导通路JNKMAPK的选择性抑制剂SP600125能促进ESAT-6刺激单核细胞IL-12p40产生,而信号通路PKR抑制剂2-AP有显著性抑制其作用。结论ESAT-6抗原诱导人THP-1单核细胞IL-12产生,JNK MAPK及PKR信号通路可能参与了此过程的调控。     

联合检测脑脊液中腺苷脱氨酶和结核杆菌DNA对结核性脑膜炎的诊断价值

目的 探讨联合检测脑脊液（CSF）腺苷脱氨酶（ADA）和荧光定量聚合酶链反应（PCR）检测结核杆菌DNA（TB-DNA）对结核性脑膜炎的早期诊断价值.方法 选择确诊的结核性脑膜炎患者（发病10天以内,未进行抗结核治疗）51例,非结核性脑膜炎患者51例,留取患者脑脊液4ml进行ADA、TB-DNA检测.结果 患者脑脊液中ADA水平[（24.88±10.97） U/L]显著高于非结核性脑膜炎组[（11.14±4.02）U/L] （P ＜0.05）,两组患者脑脊液中TB-DNA水平比较,差异有统计学意义（P＜0.05）;联合检测脑脊液中ADA、TB-DNA水平诊断结核性脑膜炎的敏感性、特异性和准确性分别为92.19％、80.39％和86.27％.结论 联合检测脑脊液中ADA、TB-DNA可以作为临床上早期诊断结核性脑膜炎的有效指标之一.     

早期分泌抗原靶6及基质金属蛋白酶9评估结核性毁损肺严重程度的应用价值

目的 探讨早期分泌抗原靶6(early secretory antigen target-6,ESAT-6)及基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)评估结核性毁损肺严重程度中的应用价值。方法 选取2018年1月至2019年1月昆明市第三人民医院收治的55例结核性毁损肺患者(A组),40例继发性肺结核患者(B组),以及同期在本院进行体检的健康志愿者20名(C组)作为研究对象,收集研究对象临床资料及实验室检查结果,比较和分析各组ESAT-6、MMP-9水平[中位数(四分位数)]及T淋巴细胞计数[中位数(四分位数)]的变化情况。结果 A组ESAT-6表达水平为28.83(19.26,38.20)pg/ml,B组为22.26(17.85,25.01)pg/ml,均明显高于C组[0.06(0.03,1.50)pg/ml],差异均有统计学意义(U=63.155,P<0.01;U=49.725,P<0.01)。A组MMP-9表达水平为12.54(9.01,18.11)μg/ml,明显高于B组的6.80(4.10,12.29)μg/ml和C组的1.14(0.72,1.29)μg/ml,差异均有统计学意义(U=26.097,P<0.01;U=66.309,P<0.01)。所有入组肺结核患者(A组和B组)ESAT-6的表达量水平为24.05(18.54,34.89)pg/ml与CD4⁺T淋巴细胞计数[476.00(312.00,647.00)个/μl]和CD3⁺T淋巴细胞计数[599.00(456.00,762.00)个/μl]呈负相关关系(r=-0.462,P<0.01;r=-0.275,P=0.003);MMP-9表达水平为11.05(6.30,14.28)μg/ml,与CD4⁺和CD3⁺T淋巴细胞计数也呈负相关关系(r=-0.499,P<0.01;r=-0.341,P<0.01)。A组中重度通气功能障碍者(52.7%,29/55)MMP-9表达水平[10.79(7.10,12.54)μg/ml]明显低于极重度通气功能障碍者(47.3%,26/55)MMP-9表达水平[17.39(12.47,33.30)μg/ml],差异有统计学意义(U=150.000,P<0.01)。结论 ESAT-6与MMP-9在肺结核患者中表达增高,与患者免疫功能相关。MMP-9水平与结核性毁损肺严重程度相关。     

早期分泌抗原靶6及免疫和炎症指标对抗结核药物性肝损伤的诊断价值

目的 分析相关免疫及炎症指标对抗结核药物性肝损伤的诊断价值。方法 选择2019年1月至2021年1月昆明市第三人民医院收治的符合入组标准的肺结核患者325例[包括发生抗结核药物性肝损伤的患者115例(A组)和未发生肝损伤的患者210例(B组)]和同期的门诊健康体检者98名(C组)作为研究对象。采用多因素logistic回归模型分析3组研究对象中免疫细胞、炎症指标、血清早期分泌抗原靶6(ESAT-6)及基质金属蛋白酶9(MMP-9)和MMP-14与发生抗结核药物性肝损伤的相关性。结果 A组患者年龄中位数(四分位数)[51.2(18.9,77.2)岁]高于B组[39.1(19.7,63.3)岁]和C组[36.3(20.1,61.3)岁],差异均有统计学意义(H=27.695、29.982,P值均=0.000);A组患者CD4⁺ T淋巴细胞计数[295.0(155.0,449.0)个/μl]低于B组[571.0(397.0,642.0)个/μl]和C组[775.0(710.0,993.0)个/μl],差异均有统计学意义(H=27.225、40.117,P值均=0.000);A组血浆降钙素原(PCT)[4.3(0.9,11.5)ng/ml]、D-二聚体[4.5(0.7,8.4)μg/ml]、ESAT-6[59.3(27.1,66.5)pg/ml]、MMP-9[29.1(18.6,39.6)ng/ml]表达水平均高于B组 [分别为2.8(0.5,8.6)ng/ml、2.3(0.5,5.1)μg/ml、32.5(25.8,59.2)pg/ml、17.2(12.7,21.3)ng/ml],差异均有统计学意义(H=28.991、29.879、32.045、31.122,P值均=0.000);A组和B组MMP-14表达量[分别为54.7(41.4,66.7)ng/ml和60.2(45.2,65.1)ng/ml]均高于C组[5.5(2.8,6.3)ng/ml],差异均有统计学意义(H=49.209、53.436,P值均=0.000)。多因素logistic回归分析显示,高龄(>65岁)、ESAT-6升高(>30.5pg/ml)是抗结核药物性肝损伤发生的独立危险因素[OR(95%CI)=11.289(4.355~24.361),P=0.000;OR(95%CI)=9.479(3.340~21.653),P=0.000]。结论 针对高龄和ESAT-6高表达肺结核患者及早使用保肝药物,可能会降低抗结核药物性肝损伤发生的风险。对CD4⁺ T淋巴细胞计数较低,降钙素原、D-二聚体和MMP-9高表达的肺结核患者也应提高警惕。     

Diagnostic Value of Early Secreted Antigenic Target-6 for the Diagnosis of Tuberculous Meningitis Patients

Background:   

The early diagnosis of tuberculous meningitis (TBM) is very crucial, since delayed diagnosis can lead to various neurological manifestations. We have previously developed an in-house indirect enzyme-linked immunosorbent assay (ELISA) for TBM diagnosis using the Antigen 85 (Ag 85) complex. It has been suggested that the Ag 85 complex might give false-positive reactions for individuals vaccinated with Bacillus Calmette-Guérin (BCG).     

Diagnostic Challenges and Dilemmas in Tuberculous Meningitis

Tuberculosis is a global burden with an unacceptably high mortality rate, especially in low- and middle-income countries. We reported the case of 34-year-old Somali female with no significant risk factors who initially presented with headache and blurred vision. The patient subsequently developed altered mental status and significant vision changes. Initial lumbar puncture showed lymphocytic pleocytosis with negative gram stain, acid-fast bacilli stain, and culture. Initial polymerase chain reaction for tuberculosis was negative. The patient worsened despite receiving broad-spectrum antibiotics. The patient had a prolonged hospital course and eventually required lumbar drain placement for hydrocephalus. Repeated polymerase chain reactions for Mycobacterium tuberculosis from the lumbar drain samples was positive, and the diagnosis of tuberculous meningitis was confirmed. The patient improved after lumbar drain placement and treatment with isoniazid, rifampin, pyrazinamide, ethambutol and steroid tapering. This case illustrated the challenge of diagnosing tuberculous meningitis.     

数字减影磁共振成像对脑膜炎的诊断价值

目的评价数字减影磁共振成像(MRI)技术对感染性脑膜炎的诊断价值。方法47例脑膜炎病人进行MRI平扫及增强扫描,然后用T1W增强后的图像与增强前的图像进行减影,比较减影前后图像对脑膜强化显示能力及对比度/噪声比值(CNS值)。根据发病后MRI检查时间分成3组,A组<3周,B组3周至3个月,C组>3个月。结果减影图像比增强后的图像更清晰、更直观,减影图像CNS值为116.14±4.98,增强图像CNS值为21.27±2.96,减影图像CNS明显高于增强图像,两者比较有统计学意义(P<0.05)。减影前不能清楚判断脑膜强化的病例,减影后能做出明确的判断。特别是对早期病例(A组),敏感性从57.1%提高到89.3%(P<0.05)。结论数字减影MRI处理技术,可提高脑膜强化显示能力,特别是对诊断早期感染性脑膜炎具有重要应用价值。     

结核性脑膜炎脑脊液中粒细胞集落刺激因子的测定

本文采用ＥＬＩＳＡ法检测２３例结核性脑膜炎及２１例非结核性脑膜炎脑脊液中的Ｇ－ＣＳＦ。结脑组（０．６５＋０．３２ＯＤ）明显高于非结脑组（０．１５±０．１１ＯＤ）（Ｐ＜０．０１）。诊断结脑的敏感性为９１．３％，特异性为９０．５％。Ｇ－ＣＳＦ在抗感染的非特异性细胞免疫中起重要作用，细菌及其产物是刺激和调节Ｇ－ＣＳＦ产生的主要物质。     

Safety of ESAT-6

A recombinant dimer of the Mycobacterium tuberculosis (MTb) 6 kDa early secreted antigenic target (ESAT-6) was produced in Lactococcus lactis. Pharmacodynamic and safety studies were carried out in guinea pigs, rats, mice and dogs with intradermal (id), subcutaneous (sc) and intravenous (iv) administration of the antigen. In contrast to tuberculin purified protein derivative (PPD) the recombinant dimer (rdESAT-6) was able to discriminate MTb infection from BCG vaccination in vivo. In guinea pigs sensitized by infection with MTb, 1 microg rdESAT-6 gave a mean delayed-type hypersensitivity (DTH) response of 22 mm, a significantly stronger reaction than in animals sensitised by the environmental mycobacteria M. kansasii, M. szulgai and M. marinum. rdESAT-6 proved to be a safe tuberculin reagent in a dose range of 1-1000 microg with no or only minor local reactions.     

降钙素原与CA125联合检测对结核性胸腔积液的诊断价值

目的探讨联合检测血清和胸腔积液降钙素原(PCT)、CA125对结核性胸腔积液的诊断价值。方法 30例结核性胸腔积液、20例肺炎旁积液、30例恶性胸腔积液,采用化学发光法测定血浆和胸腔积液中的PCT水平,快速发光免疫分析测定CA125的水平。结果结核组、肺炎组血清PCT水平差异无统计学意义(P>0.05),结核组和肺炎组胸腔积液中PCT的表达均较对照组明显升高(均P<0.05)。结核组和恶性组血清及胸腔积液中CA125水平较肺炎组显著升高(P<0.01)。结论联合检测血清及胸腔积液中PCT与CA125水平可提高结核性胸腔积液诊断的敏感性和特异性。     

小口径腹腔镜检查对结核性腹膜炎的诊断价值：21例临床分析

背景：结核性腹膜炎临床表现常不典型，误诊、漏诊率较高。目的：探讨小口径腹腔镜检查对结核性腹膜炎的诊断价值和安全性。方法：回顾性分析2006年1月～2007年12月于复旦大学附属中山医院行小口径腹腔镜检查，最终诊断为结核性腹膜炎的21例患者的一般情况、腹腔镜表现和实验室、辅助检查结果。结果：1例患者因腹腔严重黏连而终止检查；20例（95．2％）腹腔镜直视可见腹腔内弥漫粟粒样结节，结合临床表现诊断为结核性腹膜炎，其中17例（81．0％）取得活检组织，经病理检查明确诊断。21例患者中，渗出型11例（52．4％），黏连型7例（33．3％），干酪型3例（14.3％）。术后并发症发生率为9．5％，包括穿刺部位出血和皮下气肿各1例。小口径腹腔镜直视诊断阳性率显著高于结核菌素纯蛋白衍生物（PPD）试验、腹水乳酸脱氢酶（LDH）、血清和腹水肿瘤相关糖链抗原125（CA125）、腹水腺苷脱氨酶（ADA）检测等（P〈0．05）。结论：小口径腹腔镜检查用于诊断结核性腹膜炎，操作简便、安全，诊断准确性高。     

Approach to the Diagnosis and Management of Tuberculous Meningitis

Meningitis caused by Mycobacterium tuberculosis remains an important cause of morbidity and mortality worldwide, and presents particular challenges in terms of diagnosis and management. The nonspecific clinical presentation of tuberculous meningitis (TBM) has led researchers to develop newer molecular methods of making the diagnosis. Several of these methods have excellent sensitivity and specificity, although many are not yet available for clinical use. Successful treatment of TBM requires a combination of antimicrobial agents, with vigilance regarding the possibility of disease caused by resistant organisms. Adjunctive corticosteroids also have a role in treating this potentially devastating infection, as can neurosurgery. With proper therapy, morbidity and mortality can be minimized in patients with TBM.     

BKV-DNA and JCV-DNA in CSF of Patients with Suspected Meningitis or Encephalitis

Abstract. Background: Few studies have looked for the polyoma viruses JC or BK virus in the central nervous system (CNS) of patients without neurological symptoms or with neurological symptoms other than progressive multifocal leukoencephalopathy (PML). PCR-microplate hybridization method was employed for the detection of BKV-DNA or JCV-DNA in cerebrospinal fluid (CSF) specimens from patients with suspected meningitis or encephalitis. Materials and Methods: A total of 181 CSF specimens from 151 patients with suspected meningitis or encephalitis was examined for BKV or JCV using PCR-microplate hybridization method. None of the patients had (clinically diagnosed) PML. A control group consisting of 20 CSF specimens from normal subject was also included. Results: BKV DNA was found in five out of 131 (3.8%) and JCV DNA in two out of 131 (1.5%) of the patients with suspected meningitis or encephalitis by PCR ELISA. BKV or JCV DNA was not detected in CSF samples of any of 19 HIVpositive patients. BKV and JCV DNAs were detected respectively in two CSF samples in which Mycobacterium tuberculosis (TB) PCR was also positive. Another patient who was positive for JCV PCR died with a diagnosis of cerebral lymphoma. Among the BK virus infected patients there was a patient with a previous history of hemolytic uremia and acute renal failure. Neither BKV nor JCV DNA was found in any of the 20 CSF samples from normal patients undergoing lumbar puncture for myelography as a part of an investigation of lower back pain. Conclusion: These results suggest that BK virus may be associated with neurological diseases either in immunocompetent or immunocompromised patients. Detection of BKV and JCV DNA in the CSF of the patients suspected to have either meningitis or encephalitis suggests that these viruses may have an etiological role. Thus, diagnostic tests for BK and JC viruses should be included in the investigative program for meningitis or encephalitis patients.     

结核分枝杆菌早期分泌抗原靶6家族其他蛋白抗原的研究进展

目前结核分枝杆菌的致病机制和宿主的防御机制尚不十分清楚,结核分枝杆菌蛋白抗原生物学特性的研究有助于该问题的阐明,将为结核病疫苗、免疫诊断试剂和新药的开发奠定基础.早期分泌抗原靶6（ESAT-6）家族蛋白是一类小分子蛋白,呈螺旋状结构,它们通过ESAT-6分泌系统（ESAT-6 secretion system,ESX）分泌到细胞外.该家族共有23个成员（EsxA～W）,其在基因组上相邻排列的2个蛋白编码基因形成11个类操纵子结构的基因对.该家族蛋白参与宿主与致病菌之间的相互作用,是人免疫系统识别的优势抗原,大多数是T细胞优势抗原,在结核分枝杆菌致病机制和机体的免疫保护机制方面起关键作用.鉴于EsxA和EsxB的研究概况国内外报道较多,而其他ESAT-6家族成员研究报道较少.因此,笔者着重概要地介绍ESAT-6家族其他成员的国内外研究进展情况,为进一步的深入研究和应用奠定基础.