病毒性脑炎脑脊液病毒核酸检测的研究进展

目的 脑脊液病毒核酸检查是诊断病毒性脑炎的关键，而长期以来病毒学诊断技术敏感性和特异性很差，远远不能满足临床要求。随着分子生物学的迅速发展，以PCR技术为基础的各种分子生物学诊断技术，成为生物医学领域内最有价值的研究手段和病毒学诊断的金标准，已经广泛的应用于临床标本的检测，为病毒性脑炎病原学的基因诊断带来了突破性的进展；方法本文就病毒性脑炎的脑脊液采集时间、预处理和保存以及PCR技术的应用研究进展进行了综述；结论套式、逆转录和荧光定量PCR已越来越多的应用于临床，预料原位PCR将在检测脑脊液病毒核酸检测等方面有着重要的意义。     

病毒性脑炎患儿脑脊液NO含量与临床意义

目的探讨病毒性脑炎患儿脑脊液一氧化氮含量(NO)变化与脑损伤程度的相关性.方法用比色法检测37例病毒脑及20例正常对照儿脑脊液NO含量.结果病毒脑患儿脑脊液NO含量明显高于对照组 ,脑损伤重者NO含量更高,急性期的脑脊液NO含量明显高于恢复期.结论病毒脑患儿脑脊液NO含量升高,增高幅度与脑损伤程度有关,NO含量测定有助于脑脊液常规及生化正常的病毒脑的诊断.     

病毒性脑炎脑脊液病毒核酸检测的研究进展

目的脑脊液病毒核酸检查是诊断病毒性脑炎的关键,而长期以来病毒学诊断技术敏感性和特异性很差,远远不能满足临床要求。随着分子生物学的迅速发展,以PCR技术为基础的各种分子生物学诊断技术,成为生物医学领域内最有价值的研究手段和病毒学诊断的金标准,已经广泛的应用于临床标本的检测,为病毒性脑炎病原学的基因诊断带来了突破性的进展;方法本文就病毒性脑炎的脑脊液采集时间、预处理和保存以及PCR技术的应用研究进展进行了综述;结论套式、逆转录和荧光定量PCR已越来越多的应用于临床,预料原位PCR将在检测脑脊液病毒核酸检测等方面有着重要的意义。     

病毒性脑炎患儿血清,脑脊液中细胞因子测定的临床意义

对病毒性脑炎患儿血清、脑脊液中的ＩＬ－６、ＩＬ－８、ＴＮＦ－α进行了测定。结果表明：血清中的ＩＬ－６、ＩＬ－８、ＴＮＦ－α含量明显高于正常对照组；脑脊液中也有一定水平的ＩＬ－６、ＩＬ－８、ＴＮＦ－α。分析认为：这些细胞因子不仅具有抗病毒作用而且还参与了小儿病毒性脑炎的病理过程。     

一起病毒性脑炎病因的分子生物学鉴定

目的探讨2003年山东济南地区病毒性脑炎（VE）流行病因。方法通过随机PCR及肠道病毒特异性的RT—PCR扩增获得病毒分离株特异性核酸片段，测序并进行同源性分析和多序列比对，确定病毒种类及分型。结果从患者标本中分离到5株病毒，以随机PCR获得其中1株病毒的核酸片段，经BLAST分析发现其与肠道病毒同源性最高，然后分别测得5株病毒5’端非编码区及部分VP1区核酸序列，经序列比对分析，确定5株病毒均为小RNA病毒科肠道病毒属的柯萨奇B5，并且均与2002--2004年间浙江无菌性脑膜炎流行期间分离的柯萨奇B5分离株同源性最高（95％以上）。结论柯萨奇B5是2003年济南地区VE流行的重要病因之一。     

病毒性脑炎患者的脑电图分析

目的：探讨脑电图（EEG）检查在病毒性脑炎的诊断和治疗中的作用。方法：对250例病毒性脑炎的病人进行EEG检查及跟踪观察。结果：250例患者中EEG诊断界限28例,轻度异常58例,中度异常128例,重度异常40例。EEG大多为弥慢性异常。其中有44例在弥漫性异常的基础上出现局灶性改变,临床伴有抽风发作者EEG出现尖波,棘波,尖棘慢综合波等改变（异常率占90．40％）与其他文献报道相符合。结论：EEG检查对病毒性脑炎的诊断治疗及病情转归,是不可缺少的重要检查手段之一。     

应用液相芯片技术检测病毒性脑炎患者脑脊液中细胞因子的变化及临床意义

目的:探讨病毒性脑炎(VE)患者脑脊液(CSF)中细胞因子的变化及其临床意义。方法:22例VE患者和15名非中枢神经感染者作为研究对象,采用lum inex液相芯片技术检测患者CSF及血清中IL-6、IL-8、IL-10、TNFα-的水平,并行对照分析。结果:VE组CSF中IL-6、IL-8、IL-10、TNFα-水平明显高于对照组,差异均具有统计学意义;抗病毒治疗1周后VE组CSF中IL-6、IL-8及IL-10水平较治疗前下降,差异具有统计学意义,TNFα-水平与治疗前比较无明显统计学差异;VE组CSF中IL-6、IL-8、IL-10、TNFα-水平均明显高于血清中水平。结论:IL-6、IL-8、IL-10、TNFα-在VE的病理过程中颅内合成显著增加,参与VE急性期脑损伤过程,在疾病的发生、发展及转归中发挥了重要的作用。     

重症病毒性脑炎的护理对策

目的：探讨重症病毒性脑炎的护理对策。方法：对13例重症病毒性脑炎在治疗的同时加强护理，主要对精神症状、意识障碍、高颅压、高热、癫痫持续状态提出相应护理对策。结果：经过治疗和护理，痊愈9例，遗留后道症3例，死亡1例。结论：护理是重症病毒性脑炎治疗成功的重要措施。     

病毒性脑炎的脑电图分析

目的：探讨病毒性脑炎中脑电图特点及其预后价值.方法：对174例病毒性脑炎的脑电图进行跟踪观察并予回顾性分析.结果：在174例病毒性脑炎患者中133例脑电图异常,异常率为76.4％,其中轻度异常81例（46.6％）,中度异常38例（21.8％）,重度异常14例（8.0％）.大多数病毒性脑炎患者脑电图显现广泛的弥漫性或局限性异常,局限性、低波幅慢波其后遗症发生率高,以智能、运动障碍多见,而高波幅慢波其后遗症以继发性癫癎多见.结论：病毒性脑炎脑电图异常率较高,脑电图对病毒性脑炎脑功能恢复的诊断及后遗症的监测有一定价值.     

脑脊液指标在自身免疫性脑炎和病毒性脑炎鉴别诊断中的价值

目的 比较脑脊液指标在自身免疫性脑炎和病毒性脑炎中的差异,探讨其在两种不同类型脑炎鉴别诊断中的应用价值。方法 对2016年1月至2022年9月就诊于首都医科大学附属北京天坛医院的224例脑炎患者进行回顾性分析,根据临床最终诊断分为自身免疫性脑炎(autoimmune encephalitis, AE)122例,病毒性脑炎(viral encephalitis, VE)102例。分析两组患者的一般资料(年龄、性别),测定两组患者脑脊液白细胞水平(CSF-WBC)和脑脊液蛋白水平(CSF-PRO);采用速率散射比浊法分别测定血清的免疫球蛋白G(S-IgG)、白蛋白(S-Alb)和脑脊液的免疫球蛋白G(CSF-IgG)和白蛋白(CSF-Alb)水平,根据公式计算白蛋白商(QAlb)、24h鞘内IgG(24h IgG)合成率、IgG指数;采用琼脂糖凝胶等电聚焦电泳检测脑脊液中的寡克隆区带(CSF-OCB),并进行对比分析。将P<0.05的指标进行ROC曲线分析,并分析其诊断效能。结果 CSF-WBC、CSF-PRO、CSF-ALB、CSF-IgG、24h IgG合成率异常率、QAlb异常...     

以精神症状为突出表现的病毒性脑炎动态脑电图的研究

目的 研究动态脑电图对于以精神症状为突出表现的病毒性脑炎辅助诊断价值。方法 对精神症状为突出表现，临床拟诊为病毒性脑炎的病人在入院3d内分别进行常规脑电图和动态脑电图检查，其中有40例确诊为病毒性脑炎，采用配对χ^2检验（McNemerχ^2检验）比较两种检查方法对于中度或重度异常脑电活动的机率。结果动态脑电图和常规脑电图的中度或重度脑电活动的发现率分别为：80％、65％，差异有显著性（p〈0．05）；发现痫样放电的机率分别为：42．5％、5％，差异有显著性（p〈0．005）。动态脑电图检查发现40例病人的睡眠生理波都不清楚。结论 动态脑电图对以精神症状为突出表现的病毒性脑炎早期诊断有重要的辅助诊断价值，相对于常规脑电图有更高的敏感性：睡眠节律紊乱是这类病毒性脑炎的共同特征。     

Coltivirus新成员的分离和鉴定

１９９１年，在甘肃和北京地区采集蚊子，从中分离到１０株病毒，该类病毒在Ｃ６／３６，ＢＨＫ－２１及Ｖｅｒｏ细胞上可出现病变，使乳鼠发病、致死，抵抗乙醚和５－溴脱氧尿苷，电镜观察为球形、无包膜，直径约６２．７±３．１３ｎｍ。ＰＡＧＥ示该类病毒ＲＮＡ为１２节段，分为４个不同的电泳带型，均与云南分离株不同。交互中和试验和交互补体结合试验表明，中国分离株之间抗原性有变异。中国分离株与Ｃｏｌｔｉｖｉｒｕｓ属已知成员科罗拉多蜱热病毒、加利福尼亚分离株Ｓ６－１４－０３及德国分离株Ｅｙａｃｈ有血清学交叉反应（１：４～１：８），但血清型明显不同，因而，中国分离株当属Ｃｏｌｔｉｖｉｒｕｓ属的新的成员。     

单克隆抗体ELISA法快速诊断疱疹性脑炎

本文用单纯疱疹病毒单克隆抗体（ＨＳＶ－ＭｃＡｂ）ＥＬＩＳＡ法检测了１２８例病毒性脑炎患儿脑脊液中ＨＳＶ特异性抗原（ＨＳＶ—Ａｇ），２０例其他神经系统疾患也作了测定，３２例脑炎患儿同时取脑脊液和血清作ＨＳＶ—ＩｇＧ抗体水平测定比较，病毒性脑炎患儿的ＨＳＶ－Ａｇ阳性检出率为１９．５％，与血清／脑脊液的ＨＳＶ－ＩｇＧ测定比较，敏感性８４．６％，特异性９４．７％。用ＥＬＩＳＡ法检测脑脊液中ＨＳＶ－Ａｇ是一种简便、快速、敏感、特异的方法，对疱疹性脑炎具有早期诊断价值。     

用ELISA法快速检测病毒性脑炎患儿脑脊液中单纯疱疹病毒特异性抗原

用单纯疱疹病毒单克隆抗体（ＨＳＶ－ＭｃＡｂ）ＥＬＩＳＡ法检测了６２例病毒性脑炎患儿脑脊液中ＨＳＶ特异性抗原（ＨＳＶ－Ａｇ），２０例其他脑神经系统疾患者。３２例脑炎患儿同时取脑脊液和血清作ＨＳＶ－ＩｇＧ抗体水平测定比较。病毒性脑炎患儿的ＨＳＶ－Ａｇ阳性检出率为２０．６％，与血清／脑脊液的ＨＳＶ－ＩｇＧ测定比较，灵敏性８４．６％，特异性９４．７％。用ＥＬＩＳＡ法检测脑脊液中ＨＳＶ－Ａｇ是一种简便、快速、灵敏、特异的方法，对疱疹性脑炎具有早期诊断价值。     

Detection of both herpes simplex and varicella-zoster viruses in cerebrospinal fluid from patients with encephalitis

Cerebrospinal fluid (CSF) samples from 46 patients with encephalitis were studied for the presence of herpes simplex virus (HSV) types 1 and 2 and/or varicella zoster virus (VZV)-specific DNA sequences by the polymerase chain reaction (PCR) assay. Patients were studied because of detection of intrathecal production of IgG antibody to HSV alone (10 patients, Group A) or to both HSV and VZV (11 patients, Group B) or because of the presence of specific anti-HSV IgG in CSF without evidence of intrathecal antibody production (25 patients, Group C). CSF samples taken between days 1 and 10 from onset of encephalitis were available from all patients, and follow-up samples (taken after 10 days from onset) were obtained from some of them. Positive PCR results were obtained in a total of 13 patients. Four patients (three from Group A and one from Group B) gave amplification of HSV type 1 DNA alone, two patients (both from Group B) showed amplification of VZV DNA alone, and seven patients (all from Group B) gave dual amplification of both HSV type 1 and VZV DNA sequences in CSF. All CSF samples from patients in Group C were negative by PCR. Ten patients with CSF samples positive by PCR lacked a prior history of herpetic cutaneous lesions. In seven patients, serum antibody tests (specific IgM detection and specific IgG avidity assays) identified both primary and recurrent infections. The results suggest that the dual presence of IgG antibody to both HSV and VZV in CSF from patients with encephalitis may reflect in some cases a dual infection of the central nervous system caused by both agents. © 1996 Wiley-Liss, Inc.     

Cloning and Sequencing of Complete cDNA of Japanese Encephalitis Virus YL Strain in Taiwan

We determined the complete nucleotide sequence of the YL strain of Japanese encephalitis virus and its amino acid sequence was deduced. Our results displayed that the genome of YL strain contained a single open reading frame of 10,296 nucleotides (nts) which was flanked by untranslated region (UTR) containing 95 bases at the 5-end and 586 bases at the 3-end, respectively. Comparison of sequences showed that the overall amino acid sequence and 3 UTR of YL were similar to those of the virulent strain JaGAr01. However, some significant amino acid differences of viral envelope (E) protein were observed between YL and JaGAr01; the amino acid sequence of E protein in YL strain possessed RGG_(387–389) tripeptide instead of RGD_(387–389) in JaGAr01 and in other strains; and another amino acid is K₍₁₃₈₎ in YL, not E₍₁₃₈₎ found in others. These differences suggested that the YL strain impairs in viral attachment to the cell surface and loses neuroinvasiveness, and therefore this strain was used as a live attenuated vaccine.     

血清sVCAM-1、NSE与Gal-9联合检测病毒性脑炎患儿病情程度变化的判定价值

目的分析血清sVCAM-1、NSE与Gal-9联合检测病毒性脑炎患儿病情程度变化的判定价值。方法选择2016年11月至2017年11月的VE患儿116例作为研究组,按照入院时的生命体征和脑干受损和呼吸抑制情况分为重症组和轻症组,接受2周治疗后,选择病情好转为恢复期的患儿进行相关研究,选择同期在我院接受体检的健康儿50例作为对照组。比较不同组和不同病程间血清sVCAM-1、NSE与Gal-9水平。结果重症组血清sVCAM-1、NSE与Gal-9水平显著高于轻症组和对照组(P<0.05)。恢复期患儿sVCAM-1、NSE较治疗前显著降低(P<0.05),其中sVCAM-1与对照组差异无统计学意义(P>0.05),Gal-9较治疗前显著升高(P<0.05)。结论 VE患儿血清sVCAM-1、NSE和Gal-9明显升高,并且病情严重患儿高于轻症患儿,处于恢复期的患儿的sVCAM-1、NSE显著降低而Gal-9明显升高。     

Monitoring of herpes simplex virus DNA types 1 and 2 viral load in cerebrospinal fluid by real‐time PCR in patients with herpes simplex encephalitis

A quantitative polymerase chain reaction (PCR) assay was evaluated retrospectively on 92 cerebrospinal fluid (CSF) samples from 29 patients with herpes simplex virus (HSV) encephalitis with the aim to study if the concentration of HSV genomes can be used as a prognostic marker and for monitoring of antiviral therapy. The results were compared to those obtained previously by nested PCR, and the numbers of HSV genomes/ml were evaluated in correlation to patient outcome and treatment. The aims were to compare the sensitivity of a conventional nested PCR to a quantitative PCR, to investigate the range of HSV genome concentration in initial samples and to evaluate possible relationships between the HSV DNA concentrations in CSF, neopterin levels, and outcome of disease. The 29 initial samples contained between 2 × 10² and 42 × 10⁶ HSV genomes/ml. There was no apparent correlation between the amount of HSV DNA in the initial samples and income status, initial neopterin levels, or prognosis. The number of HSV genomes/ml declined after treatment in all patients, but HSV DNA was still detectable after day 20 in 3 out of 16 patients. A long duration of genome detectability was found to correlate with poor outcome. There was no difference in sensitivity between the nested PCR and the quantitative PCR. While the quantitative PCR is more rational than a nested PCR, the quantitation of HSV genomes does not seem very useful as a prognostic marker in HSV encephalitis. J. Med. Virol. 81:1432–1437, 2009. © 2009 Wiley‐Liss, Inc.     

Correlation of tumor necrosis factor levels in the serum and cerebrospinal fluid with clinical outcome in Japanese encephalitis patients

To investigate the prognostic role of tumour necrosis factor (TNF) in Japanese encephalitis virus (JEV) infection, we measured the immunoreactive forms of TNF concentrations in the serum and cerebrospinal fluid (CSF) of 47 laboratory-confirmed cases of JE. It was observed that TNF levels were elevated (>15 pgm/ml) in all the 47 serum samples (range 19.4–923.8 pg/ml), while in 46/47 CSF samples TNF was elevated (range 10.8–376 pg/ml). The mean (SD) TNF levels in the serum of fatal cases was 234.34 pg/ml (304.40) as compared to the mean of 85.31 pg/ml (SD 153.92) in nonfatal cases. Similar observations were also made with respect to the TNF levels in the CSF; the mean of fatal cases was 69.39 pg/ml (SD 39.00) in contrast to the mean of 62.41 pg/ml (SD 75.25) of nonfatal cases. The increase in TNF levels did not show any correlation to the duration of illness. It was further observed that the mortality rate increased with increasing concentrations of TNF in the serum and CSF. Correlation of laboratory parameters to final outcome revealed that TNF concentrations above 50 pg/ml in serum correlated significantly (P = .05) with a fatal outcome, whilst high levels of JEV-IgM antibodies (>500 units) in the CSF correlated with a nonfatal outcome (P = .03). These results suggest that TNF can be used as a possible prognosticator of a fatal outcome in JEV infection. J Med Virol 51:132–136, 1997. © 1997 Wiley-Liss, Inc.