单纯疱疹病毒性脑炎诊断方法与治疗观察

目的探讨单纯疱疹病毒性脑炎（ＨＳＶＥ）的诊断方法，了解对ＨＳＶＥ的治疗效果。方法应用聚合酶链反应（ＰＣＲ）技术和酶联免疫吸附测定（ＥＬＩＳＡ）方法，对２７例病毒性脑炎患儿的脑脊液进行单纯疱疹病毒（ＨＳＶ）ＤＮＡ和特异性ＩｇＭ抗体检测。结果ＨＳＶＤＮＡ阳性１１例；ＨＳＶＩｇＭ阳性４例，其中Ⅰ型３例，Ⅱ型１例；ＤＮＡ与ＩｇＭ同时阳性３例。将脑脊液检查ＨＳＶＤＮＡ阳性或ＨＳＶＩｇＭ抗体阳性者１２例诊断为ＨＳＶＥ，占４４％。对ＨＳＶＥ患儿予以静脉滴注无环鸟苷，连续用药一周，结果治愈８例，好转２例，自动出院及死亡各１例。结论ＰＣＲ与ＩｇＭ抗体检测两种方法相结合有助于ＨＳＶＥ病原学早期诊断，并指导ＨＳＶＥ的临床治疗     

单纯交疹病毒性脑炎诊断方法与治疗观察

目的 探讨单纯疱疹病毒性脑炎（ＨＳＶＥ）的诊断方法，了解对ＨＳＶＥ的治疗效果。方法应用聚合酶链反应（ＰＣＲ）的技术和酶联免疫吸附测定（ＥＬＩＳＡ）方法，对２７例病毒性脑炎患儿的脑脊液进行单纯疱疹病毒（ＨＳＶ）ＤＮＡ和特异性ＩｇＭ抗体检测。结果 ＨＳＶ－ＤＮＡ阳性１１例，ＨＳＶ－ＩｇＭ阳性４例，其中Ⅰ型３例，Ⅱ型１例；ＤＮＡ与ＩｇＭ同时阳性３例。将脑脊液检查ＨＳＶ－ＤＮＡ阳性或ＨＳＶ－Ｉdisplay stat     

31例散发性脑炎的病原学诊断及临床分析

经酶联免疫吸附试验（ＥＬＩＳＡ）检测３１例散发性脑炎患儿血清和脑脊液（ＣＳＦ）中的单纯疱疹病毒Ⅰ、Ⅱ型（ＨＳＶ－Ｉ，ＨＳＶ－Ⅱ）ＩｇＭ、ＩｇＧ抗体；用聚合酶链反应（ＰＣＲ）检测ＣＳＦ中ＨＳＶ－ＤＮＡ。结果１５例确诊为单纯疤疹脑炎（ＨＳＥ），１６例为非单纯疱疹脑炎（ＮＨＳＳＥ）。在ＨＳＥ的临床表现中，意识障碍发生率显著高于ＮＨＳＳＥ；实验室脑脊液常规检测，ＨＳＥ多见红细胞；脑电图及ＣＴ检查额，颞部位有特征性改变。提示ＨＳＥ的病原学诊断对于早期诊断及指导临床治疗具有重要意义．     

病毒性脑炎患儿肠道病毒特异性IgM抗体快速检测

目的　为了证实固相反向免疫吸附试验（ＳＰＲＩＳＴ）是肠道病毒特异性ＩｇＭ 抗体的快速检测方法。方法　用ＳＰＲＩＳＴ检测６２ 例临床拟诊为病毒性脑炎患儿脑脊液（ＣＳＦ） 或ＣＳＦ 与血清中肠道病毒ＥＣＨＯ３ 、ＥＣＨＯ５、ＥＣＨＯ７ 、Ｃｏｘ Ｂ３ 特异性ＩｇＭ 抗体。结果　１７ 例（２７－４ ％ ）ＣＳＦ阳性,ＥＣＨＯ３ 、ＥＣＨＯ５、ＥＣＨＯ７ 和ＣＯＸＢ３ 的阳性数分别为５ 例、７ 例、１ 例和４ 例。结论　ＳＰＲＩＳＴ 快速检测肠道病毒特异性ＩｇＭ 抗体可用于中枢神经系统病毒感染的早期诊断,能够指导临床治疗,避免了滥用抗生素。     

60例人类微小病毒B19感染患儿的病原血清学检测及特征

目的了解人类微小病毒Ｂ１９（ｈｕｍａｎｐａｒｖｏｖｉｒｕｓ，Ｂ１９）在儿童中的感染情况。方法采用聚合酶链反应（ＰＣＲ）和酶联免疫吸附（ＥＬＩＳＡ）方法，对１９４例住院治疗（大部分来自血液病房）患儿和１００例健康查体儿童的血清标本进行了检测。抗原为作者采用基因工程方法制备的重组Ｂ１９病毒外壳蛋白ＶＰ１和ＶＰ２。结果在１９４份患儿血清标本中，５５份检测出Ｂ１９病毒ＤＮＡ，３０份Ｂ１９病毒特异性ＩｇＭ抗体检测为阳性，３７份Ｂ１９病毒特异性ＩｇＧ抗体检测结果为阳性，阳性率分别为２８．４％，１５．５％和１９．１％，共有６０例患儿存在Ｂ１９病毒的近期感染。在１００份健康查体儿童血清标本中，３份检出Ｂ１９病毒ＤＮＡ，２份Ｂ１９特异性ＩｇＭ抗体检测结果为阳性，１２份Ｂ１９特异性ＩｇＧ抗体检测结果为阳性，阳性率分别为３０％，２０％和１２０％。结论人类微小病毒Ｂ１９在我国儿童中有较高的感染率，能够导致人类多种疾病，应该引起足够的重视     

单纯疱疹病毒性脑为临床诊断与治疗观察

为探讨单纯疱疹病毒性脑炎（ＨＳＶＥ）的诊断，应用间接ＥＬＩＳＡＳ法测定７０例病毒性脑炎患儿脑脊液的ＩｇＭ、ＩｇＧ，同时对１５例患儿的血清进行ＨＳＶ特异性ＩｇＧ的检测；采用ＥＬＩＳＡ双抗夹心法对７０例患儿的脑脊液同时检测ＨＳＶ抗原。结果：脑脊液ＨＳＶ抗原最阳性２０例，ＩｇＭ阳性７８例，ＩｇＧ血清／脑脊液比值〈２０为１２例。对ＨＳＶＥ患儿予以连续静脉滴注无环鸟苷７天，结果治愈，好转２０例（８０％）。提     

低出生体重儿TORCH先天性感染及影响因素的研究

２８１例出生体重＜２５００ｇ的早产儿和小于胎龄儿与母配对，用ＥＬＩＳＡ法和ＩＨＡ法分别检测ＣＭＶ、ＲＶ、ＨＳＶ－Ⅱ特异性ＩｇＭ抗体和Ｔｏｘ地特异性ＩｇＧ抗体。结果表明：ＴＧＯＲＣＨ先天性感染率早产儿组２７．５％，ＳＧＡ组为２０．１％，均明显高于对照组足月儿５．２％。     

PCR管单法快速检测单纯疱疹病毒性脑炎的临床价值

为了确立ＰＣＲ单管法快速检测单纯疱疹病毒（ＨＳＶ－１）的临床价值，本文对１８例经临床、ＥＥＧ，ＭＲＩ或ＣＴ诊断的单疱病毒脑炎（ＨＳＥ）患者２４份ＳＣＦ标本，先经解偶联裂解吸附预处理，抽提模板ＤＮＡ，然后进行ＰＣＲ一次性扩增研究。首次扩增１８份ＣＳＦ标本，其中１７份ＰＣＲ阳性，１份阳性，但６天后复查转为阳性。５例ＰＣＲ阳性患儿经正规抗病毒治疗后均转为阴性。对照组３０例其他中枢神经系统疾病均为阴性。结     

输血后丙型肝炎抗C22,C33C和NS5IgG及其亚群的检测和意义

为探讨抗丙型肝炎病毒（ＨＣＶ）ＩｇＧ型抗体及其亚群在ＨＣＶ感染过程中的作用，采用酶联免疫吸附法对６６例输血后ＨＣＶ感染患儿的９２份急、慢性期血清和３２份α干扰素（αＩＦＮ）治疗后血清进行抗Ｃ２２、抗Ｃ３３Ｃ和抗ＮＳ５及其亚群的检测。结果表明，急性期抗Ｃ２２和抗Ｃ３３Ｃ检出率高，分别为９８．４３％和１００．００％；而抗ＮＳ５在急性期检出率仅为５３．１２％，明显低于慢性期（９１．６７％）和同期抗Ｃ２２和抗Ｃ３３Ｃ检出率（分别为９６．８８％和１００．００％）。三种抗体ＩｇＧ亚群在急性期的检出率和抗体水平均显著低于慢性期。抗Ｃ２２ＩｇＧ１为该抗体的优势亚群；而抗Ｃ３３Ｃ无明显优势亚群；抗ＮＳ５ＩｇＧ１及ＩｇＧ３至慢性期才显示其优势。６６例患儿中，１３例接受αＩＦＮ治疗，其中５例复发患儿在ＨＣＶＲＮＡ再次出现时，抗Ｃ２２ＩｇＧ１呈现有意义的下降，而抗Ｃ２２总水平并无明显变化，而接受αＩＦＮ治疗４例有效和另４例无效患儿无类似变化。提示抗Ｃ２２和抗Ｃ３３Ｃ是诊断急性ＨＣＶ感染的敏感指标；抗Ｃ２２ＩｇＧ１可能有助于评价αＩＦＮ治疗ＨＣＶ感染的疗效。     

套式聚合酶链反应加限制酶分析检测母婴巨细胞病毒感染

为评价套式聚合酶链反应（套式ＰＣＲ）加限制酶分析在孕妇巨细胞病毒感染及其母婴宫内传播检测中的应用，采用套式ＰＣＲ加限制酶分析，病毒分离、电镜观察和特异性抗体测定，对各孕期孕妇外周血，脐血及死胎组织进行人巨细胞病毒（ＨＣＭＶ）检测。结果：３６７名孕妇ＨＣＭＶ阳性检出率为５．５％，其中，套式ＰＣＲ检出率（４．９％）高于病毒分离（３．０％，Ｐ＜０．０５）。６份ＨＣＭＶＤＮＡ阳性母血中，３份配对脐血ＨＣＭＶＤＮＡ也阳性，母－脐传播率为３／６。３对被证实为母－婴宫内传播ＨＣＭＶ的标本中，２对套式ＰＣＲ，病毒分离及特异性ＩｇＭ、ＩｇＡ均阳性，１对套式ＰＣＲ、病毒分离、特异性ＩｇＡ阳性，ＩｇＭ阴性。提示：套式ＰＣＲ能提高诊断ＨＣＭＶ的特异性与敏感性，对孕妇及胎儿／新生儿ＨＣＭＶ感染的研究有重要意义。     

新生儿中枢神经系统单纯疱疹病毒感染的临床横断面研究

目的通过分子生物学方法了解新生儿中枢神经系统单纯疱疹病毒(HSV)感染情况,并对其临床特点进行分析。方法收集考虑中枢神经系统病毒感染新生儿40例的脑脊液(CSF)标本,通过套式PCR检测CSF中HSV-DNA及酶联免疫吸附法检测CSF中特异性HSV-IgM抗体。结果2例患儿检测CSF中HSV-1 DNA( ),其母亲妊娠期均体健,无生殖器疱疹史,无皮损;1例提示为播散性感染,另1例局限为中枢神经系统感染。40例CSF检测HSV-2 DNA均阴性。结论HSV感染占新生儿中枢神经系统病毒感染的5%,不是常见病原;1型可能为新生儿HSV中枢感染的常见类型,可能与我国孕母HSV-2血清感染率较低有关。     

Herpes simplex virus infections of the central nervous system

Herpes simplex virus (HSV) infections of the central nervous system (CNS) can occur within weeks after birth (neonatal HSV disease) or in childhood or adulthood [herpes simplex encephalitis (HSE)]. Most cases of neonatal HSV disease are caused by HSV type 2, whereas virtually all cases of HSE are caused by HSV type 1. Diagnostic advances made during the past decade include the application of polymerase chain reaction (PCR) technology to cerebrospinal fluid from patients with suspected HSV CNS disease to evaluate for the presence of HSV DNA. Although not foolproof, PCR is a powerful diagnostic tool that has supplanted brain biopsy as the modality of choice for diagnosing HSV CNS disease, in no small part because of the invasiveness of brain biopsy. PCR also can provide information regarding the therapeutic response to antiviral therapy. Efforts made during the past decade to improve the outcome of HSV CNS disease have focused on increased doses of intravenous acyclovir administered for longer durations of time. Although advances have been achieved, morbidity and mortality rates from neonatal HSV disease and HSE remain unacceptably high.     

小儿病毒性脑炎的病原研究

目的　 探讨小儿病毒性脑炎的病原。 方法 　应用酶联免疫吸附方法 (ELISA)测定 5 1例病毒性脑炎患儿的脑脊液 (CSF)和血清中多种病毒特异性IgM抗体。 结果 　 5 1例病毒性脑炎患儿CSF中病毒IgM抗体阳性 2 1例 ( 4 1 2 %) ,血清中病毒IgM抗体阳性 18例 ( 35 3%)。 结论 　本地区病毒性脑炎多由肠道病毒和疱疹病毒引起 ,其中以CoxB和HSV为主要病原。     

小儿中枢神经系统单纯疱疹病毒感染的临床横断面研究

目的了解小儿中枢神经系统单纯疱疹病毒(HSV)感染的患病情况,并对其临床特点进行分析。方法收集2001年6月~2002年6月住院的中枢神经系统病毒感染患儿150例的脑脊液(cerebrospinal fluid,CSF)标本,应用套式PCR检测脑脊液中HSV-DNA及酶联免疫吸附法检测脑脊液中特异性HSV-IgM抗体。结果6例患儿脑脊液中HSV1-DNA( ),另1例HSV1-IgM( ),单纯疱疹病毒占中枢神经系统病毒感染的4.67%;呈散发性起病,无明显季节、年龄、性别分布特点,与其他病毒感染相比,惊厥持续状态、精神症状发生率高(P<0.01),意识障碍、病死率差异无显著性。结论①单纯疱疹病毒感染占儿童中枢神经系统感染的4.67%,不是儿童病毒性脑炎的常见病原,呈散发性起病,无明显季节、性别、年龄分布特点;②儿童中枢神经系统单纯疱疹病毒感染多为HSV1的原发感染,可导致脑炎、脑干脑炎、急性播散性脊髓膜炎;③单纯疱疹病毒脑炎(herpes si mplexvirus encephalitis,HSE)起病较危重,出现精神症状较多,及时有效的抗病毒治疗能明显改善病情,降低病死率。     

Non-invasive method for early diagnosis of herpes simplex encephalitis.

For the early diagnosis of herpes simplex encephalitis IgG and IgM antibodies to herpes simplex virus in cerebrospinal fluid were measured by an enzyme linked immunosorbent assay (ELISA) and a local production index was calculated. Using these three criteria, 31 cases of various neurological illnesses were analysed. All eight cases of herpes simplex encephalitis were diagnosed correctly in the acute phase, and there were no false positive results.     

Rapid diagnosis of herpetic encephalitis in children by PCR-microarray technology for simultaneous detection of seven human herpes viruses

The aim of the study was to evaluate retrospectively the usefulness of polymerase chain reaction (PCR)-microarray technology, which can simultaneously detect seven human herpes viruses for rapid and accurate diagnosis of herpetic encephalitis in children. We simultaneously amplified herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2); varicella-zoster virus; Epstein–Barr virus (EBV); cytomegalovirus (CMV); and human herpes virus 6 (HHV-6A and HHV-6B) by multiplex PCR, and genotyped by DNA microarray technology. The multiplex primers and oligonucleotide probes were designed and synthesized based on the highly conserved regions of the DNA polymerase gene in human herpes viruses. Two hundred ninety cerebrospinal fluid (CSF) specimens from children with clinical suspicion of viral encephalitis were screened by PCR-microarray technology. The results were compared with those of TaqMan PCR kits of common herpes virus. The PCR-microarray technology could detect as few as 10 copies of viral loads. There was no nonspecific hybridizing signal between probes and no cross-reaction to DNA extracted from the pathogens we used. Of 290 cases, 11 were tested positive by PCR-microarray technology. Among them, three were positive for HSV-1, two were positive for HSV-2, one was positive for EBV, two were positive for CMV, two were positive for HHV-6A, one was positive for HHV-6B, and one showed mixed infection of HSV-2 and CMV, and the positive rate was 3.8%. Compared with the results of TaqMan PCR, the sensitivity of PCR-microarray technology was 91.7%, the specificity was 100%, and the index of accurate diagnosis was 0.917. None of the 30 control CSF specimens was tested positive in both methods. Our study suggests that the simultaneous detection of seven human herpes viruses by PCR-microarray technology is the method of choice for rapid, accurate, and specific etiological diagnosis of herpetic encephalitis in children.     

小鼠单纯疱疹病毒性脑炎模型的建立

目的　探讨单纯疱疹病毒1引起小鼠单纯疱疹病毒性脑炎的发病机制。方法　4周龄雄性Balb/c小鼠和昆明小鼠各50只,通过尾静脉和颅内分别接种单纯疱疹病毒1。结果　颅内接种病毒组可导致Ballb/c小鼠和昆明小鼠体重不增、抽搐、死亡。脑组病理切片经HE染色可发现有出血、坏死、软化等改变。脑组织匀浆用PCR方法检测单纯疱疹病毒1均阳性。尾静脉接种单纯疱疹病毒1后Balb/c小鼠和昆明小鼠体重均有增加,没有抽搐等症状,小鼠存活时间均超过7天。结论　Balb/c小鼠和昆明小鼠均可感染单纯疱疹病毒1,造成单纯疱疹病毒脑炎模型,颅内接种病毒可短期内引起单纯疱疹病毒性脑炎发病,PCR检测方法是诊断单纯疱疹病毒脑炎的有效手段     

Neonatal herpes simplex virus infections: HSV DNA in cerebrospinal fluid and serum.

AIM: To investigate the diagnostic potential of herpes simplex virus (HSV) DNA in cerebrospinal fluid and serum; to correlate the findings with outcome in the child and with type of maternal infection. METHODS: Cerebrospinal fluid and serum specimens from 36 children with verified neonatal HSV infections, diagnosed between 1973 and 1996, were examined using the polymerase chain reaction technique (PCR). RESULTS: In 21 children for whom both cerebrospinal fluid and sera were available, HSV DNA was found in one or both specimens in 19 (90%). Overall, HSV DNA was found in the cerebrospinal fluid of 74% of 27 children, and in the sera of 20 out of 30 children (67%). In two children HSV DNA was not demonstrable in either serum or cerebrospinal fluid. In sequential specimens from four children, the persistence of HSV DNA after the end of intravenous treatment was associated with a poor prognosis. CONCLUSIONS: These findings indicate that HSV DNA detection in CSF and serum is highly sensitive for the diagnosis of neonatal HSV infections but does not replace the detection of virus in other locations using virus isolation and antigen detection.     

Herpes Simplex Virus Type 1 (HSV-1) and Varicellazoster Virus (VZV) Infections in Saudi Arabia

The seroepidemiology of infection due to herpes simplex type 1 (HSV-1) and varicella-zoster (VZV) viruses was investigated in 224 Saudi children aged from under 1 year to 15 years and 452 adults (healthy male blood donors and pregnant women) using a presently available sensitive indirect immunofluorescence technique to detect antibodies to these viruses in order to determine the age of primary infection. Age-specific prevalence of IgG antibodies to HSV-1 and VZV showed a progressive increase with age in both males and females with no obvious sex-related variation in the level. The overall prevalence of antibodies was 60 per cent for HSV-1 and 68 per cent for VZV in children whereas about 90 per cent of the adults showed the presence of antibodies to both viruses. Virological and serological confirmation of two cases in children of herpes simplex encephalitis (HSE) due to HSV-1 and VZV reactivation in two adults is described.     

Herpes simplex encephalitis: MRI findings in two cases confirmed by polymerase chain reaction assay

Herpes simplex virus (HSV) type I causes a fulminant necrotising meningoencephalitis distinguished from other encephalitides by its focal and often haemorrhagic nature. Specific antiviral therapy with acyclovir can significantly improve the prognosis. We present MRI findings of two cases of herpes simplex encephalitis (HSE) confirmed by PCR analysis, focusing on the serial changes after acyclovir therapy: gyral swelling, high signal intensity on T2-weighted images in the subfrontal region, temporal lobe and insula in the initial stage, then regional extension with enhancement and haemorrhage despite appropriate acyclovir therapy, and finally encephalomalacia and brain atrophy. Received: 3 October 2000 Accepted: 20 March 2001