实时RT-PCR检测159例手足口病患儿样本中肠道病毒的结果分析

目的 了解2009年4-8月首都儿科研究所附属儿童医院手足口病患儿肠道病毒的感染状况,为临床诊治提供参考.方法 采集首诊手足口病159例患儿的咽拭子和疱疹液标本,以肠道病毒(EV)通用型、柯萨奇病毒A16(CA16)型、肠道病毒71(EV71)型核酸检测试剂盒,应用实时RT-PCR法检测标本中的肠道病毒.选取阳性标本扩增VP1区,产物进行序列测定和分析.结果 (1)EV、CA16、EV71的阳性病例数分别为152、102、43;阳性率为95.6%、64.2%、27.0%.(2)CA16占EV阳性的67.3%,EV71占EV阳性的28.3%,非CA16和EV71的EV病例7例,占EV阳性的4.6%.CA16:EV71为2.37:1.(3)部分阳性标本经测序验证与此法结果一致.结论 2009年我院手足口病患儿以EV71和CA16感染为主,EV71感染的手足口病比例较2007年出现明显上升.     

小儿病毒性脑炎基因分型在诊断中应用的探讨

目的 探讨一种新的肠道病毒分型诊断方法的临床应用价值.方法 选取我院2006.06～2009.08期间诊断为病毒性脑炎的住院患儿共50例,均于入院的当天抽取脑脊液.试验先经种属特异性引物扩增,筛选出肠道病毒样本,然后分别用测序VP1及VP2序列的方法进行分型.分型结果最后经回顾性中和试验进行确认.试验同时设立阳性对照及阴性对照.结果 50例临床样本中,共有45例临床样本检测出EV RNA.其中,经VP1序列分型方法可成功扩增分型40例,反应产物为357 bp的片段.经VP2序列分型方法可成功扩增分型38例,扩增产物为584 bp的片段.微量中和试验及BLAST比对结果显示:两者在分型结果上有较高的一致性及互补性.结论 ①经VP1序列分型方法可以成功分型临床常见肠道病毒中的大多数血清型.②联合VP2序列分型方法较单用VP1序列分型方法,分型率及敏感性更高,值得临床上进一步推广.     

小儿病毒性脑炎基因分型在诊断中应用的探讨

目的 探讨一种新的肠道病毒分型诊断方法的临床应用价值.方法 选取我院2006.06～2009.08期间诊断为病毒性脑炎的住院患儿共50例,均于入院的当天抽取脑脊液.试验先经种属特异性引物扩增,筛选出肠道病毒样本,然后分别用测序VP1及VP2序列的方法进行分型.分型结果最后经回顾性中和试验进行确认.试验同时设立阳性对照及阴性对照.结果 50例临床样本中,共有45例临床样本检测出EV RNA.其中,经VP1序列分型方法可成功扩增分型40例,反应产物为357 bp的片段.经VP2序列分型方法可成功扩增分型38例,扩增产物为584 bp的片段.微量中和试验及BLAST比对结果显示:两者在分型结果上有较高的一致性及互补性.结论 ①经VP1序列分型方法可以成功分型临床常见肠道病毒中的大多数血清型.②联合VP2序列分型方法较单用VP1序列分型方法,分型率及敏感性更高,值得临床上进一步推广.     

西安地区2008年肠道病毒71型基因特征分析

目的 研究西安地区2008年引起手足口病病原构成及EV71的基因特征.方法 采集124例临床诊断手足口病病例标本,RT-PCR检测肠道病毒血清型别;挑选EV71阳性标本进行病毒分离,扩增7株EV71病毒,扩增其VP1区,测序并与EV71各血清型代表株序列比对,进行进化分析.结果 2008年西安地区手足口病（HFMD）的病原中CA16占49.45%,EV71占30.76%,其他肠道病毒占19.78%.7株EV71 VP1区与标准株序列比对,亲缘进化分析显示本地区EV71与中国大陆其他地区毒株相似.结论 2008年西安地区引起手足口病的病原以CA16为主,而EV71属于C4亚型.     

感染肠道病毒71型尸检病例的分子病原学诊断

目的 探讨感染肠道病毒71型(EV71)的尸检病例石蜡包埋组织病原学分子检测的应用价值.方法 尸体解剖2例怀疑因EV71感染死亡的患儿,对脑组织进行了组织学观察和免疫组织化学EnVision法标记;应用逆转录聚合酶链反应检测尸检后的石蜡组织中肠道病毒的核酸并测序分析.结果 2例均有中枢神经系统脑干脑炎的病理学特点.坏死神经元周围可见大量的小胶质细胞(CD68阳性)和少量的中性粒细胞(CD15阳性)浸润.2例延髓石蜡组织中均检测出EV71的核酸序列,与GenBank最新公布的安徽阜阳暴发EV71感染的病毒株序列同源性为100%.结论 从甲醛固定、石蜡包埋的EV71感染患儿脑组织中检测出病毒特异性核酸序列,及时明确诊断临床误诊、漏诊的死亡病例,深化认识了手足口病,为公共卫生管理机构及时加强EV71病原学监测提供决策依据.     

2020年西藏自治区0～5岁健康儿童肠道病毒的流行和分布

目的研究中国西藏自治区2020年0～5岁健康儿童中人肠道病毒的流行和分布。方法粪便标本取自2020年中国西藏自治区574名0～5岁的健康儿童。使用实时荧光聚合酶链反应(polymerase chain reaction, PCR)检测粪便标本中的肠道病毒核酸。对肠道病毒核酸阳性的标本, 采用逆转录PCR扩增肠道病毒VP1区并进行序列测定和分析, 根据测定的序列进行肠道病毒分子分型。结果 574例样本中检出肠道病毒(enterovirus, EV)核酸阳性89份(15.98%), EV-B检出最多, 检出率为10.98%, EV-A和EV-C检出率分别为3.66%和0.87%, 未发现EV-D。共鉴定出17种血清型, 主要血清型为E11(7.67%), 其次为EV-A71(1.74%)。各年龄组感染率存在一定的差异, 呈现为随着年龄的增大感染率逐渐降低的现象, 0岁和1岁组感染率分别为23.4%和25.0%, 而在5岁组感染率降为最低, 仅为5.8%。结论西藏自治区健康儿童粪便样本EV病原谱中以EV-B为主。序列分析提示了西藏自治区EV流行的广泛性及多样性, 以EV-A71和E11为代表...     

22例病毒性脑炎的临床表现与病毒学检测

２２例病毒性脑炎的临床表现与病毒学检测庞高龙李广武刘慧江玉卿曲静１９９６年５～９月间，我科收治了２２例临床诊断为病毒性脑炎的患儿，其临床表现与病毒学检查结果如下。２２例患儿中男１３例、女９例；１～３岁７例、～６岁６例、～１４岁９例；病例分布为５月３例...     

北京地区儿童急性呼吸道感染与肠道病毒关系探讨

目的探讨肠道病毒（enterovirus，EV）在北京地区儿童急性呼吸道感染（acute respiratory infection，ARI）中的关系及月份、年龄和性别分布特点。方法选择402例ARI患儿，取其鼻咽深部分泌物，用逆转录聚合酶链反应（RT-PCR）方法检测EV。对检测出EV阳性的患儿进行月份分布、年龄分布和性别分布的统计分析。结果402例标本中检出EV阳性共70例，阳性率为17．4％。下呼吸道感染EV阳性率为17，7％，上呼吸道感染EV阳性率为15．9％。EV在不同月份阳性率为0～36．1％，以2004年5月份阳性率最高（36．1％），2005年12月份阳性率较低（4，3％）。除12岁以上组患儿未检测到EV阳性外，在各年龄组患儿中的EV阳性率为14．8％～21．9％，4～6岁组阳性率最低，7个月至1岁组阳性率最高。EV感染的男性患儿占男性病例的16．2％，女性患儿占女性病例的19，7％。结论住院ARI患儿中，EV在下呼吸道感染中占有重要位置；春末至秋季EV阳性率较高，冬季较低；12岁以下的ARI患儿中各年龄段EV感染普遍存在；性别上无统计学意义。     

163例小儿病毒性脑炎临床分析

目的 分析小儿病毒性脑炎的临床特点和诊治.方法 对我科2008年2月～2010年9月入院治疗的163例病毒性脑炎患儿病史、体征、辅助检查、诊治经过作回顾性分析.结果 病毒性脑炎经综合治疗后大多数预后良好,在早期诊断和规范治疗的患儿更明显;但部分患儿出现各种神经系统后遗症,甚至死亡,多见于诊治较晚的患儿.结论 病毒性脑炎临床表现以发热、头疼、呕吐为主,脑脊液、脑电图和头颅MRI有助于早期诊断,而早期诊断治疗有助于提高抢救成功率.     

肠道病毒71型IgM抗体检测在手足口病早期诊断中的价值

目的 评估人肠道病毒71型(EV 71)IgM抗体在手足口病早期诊断中的价值.方法 自发病后连续每日采集2010年我院收治的38例手足口病患儿血清及咽拭子标本,分别检测EV 71IgM抗体、肠道病毒核酸、EV 71特异核酸.结果 38例手足口病患儿IgM抗体,按发病天数累加阳性率分别为:第1天60.5%、第2天71.1%、第3～4天81.5%、第5天92.1%、第6天92.1%;肠道病毒核酸阳性率为73.6%;EV 71特异核酸阳性率为60.5%.结论 EV 71 IgM抗体在手足口病发病的第1天即可出现,至第5天阳性率达到峰值,可作为手足口病早期诊断指标之一.     

儿童病毒性脑炎脑电图分析

目的：分析探讨脑电图作为儿童病毒性脑炎的早期诊断的作用。方法：对200例病毒性脑炎患儿的脑电图、脑脊液及头颅CT检查结果进行分析。结果：本组患儿中，脑电图异常率100％，脑脊液异常率52．4％，头颅CT异常率10．8％。结论：病毒性脑炎的脑电图改变较脑脊液及头颅CT的变化为早，对病毒性脑炎的早期诊断有重要意义。     

Genetic susceptibility to herpes simplex virus 1 encephalitis in mice and humans

PURPOSE OF REVIEW: Herpes simplex encephalitis is a rare complication of herpes simplex virus 1 infection that strikes otherwise healthy individuals. Its pathogenesis has long remained elusive. We highlight the investigations dealing with the genetic basis of herpes simplex encephalitis in mice and humans. RECENT FINDINGS: Mouse models have revealed the impact of various host genes on protective immunity to herpes simplex encephalitis through strain-dependent variability (forward genetics) and via targeted knockouts (reverse genetics). These studies established in particular the crucial role of IFNalpha/beta in immunity to herpes simplex virus 1, paving the way towards the elucidation of the genetic cause of human herpes simplex encephalitis. Two children with rare, specific STAT1 or NEMO mutations displayed a broad impairment of IFNalpha/beta and IFNlambda-mediated immunity and predisposition to several infectious diseases including herpes simplex encephalitis. In contrast, children with UNC93B1 and TLR3 mutations displayed a selective impairment of dsRNA-induced IFNalpha/beta and IFNlambda production and predisposition to isolated herpes simplex encephalitis. SUMMARY: Herpes simplex encephalitis results from a series of monogenic primary immunodeficiencies that impair the TLR3 and UNC-93B-dependent production of IFNalpha/beta and IFNlambda in the central nervous system, at least in a fraction of children. This is not only crucial for the understanding of immunity to herpes simplex virus 1, but also for the diagnosis and treatment of herpes simplex encephalitis.     

La Crosse encephalitis in children

BACKGROUND: La Crosse encephalitis is a mosquito-borne disease that can be mistaken for herpes simplex encephalitis. It has been reported in 28 states but may be underrecognized. METHODS: We investigated the manifestations and clinical course of La Crosse encephalitis in 127 patients hospitalized from 1987 through 1996. The diagnosis was established by serologic testing for IgM and IgG antibodies to La Crosse virus. Data were collected by chart review. RESULTS: Most of the patients were school-aged children (mean [+/-SD] age, 7.8+/-3.5 years; range, 0.5 to 15.0). Symptoms included headache, fever, and vomiting (each in 70 percent or more of the patients), seizures (in 46 percent), and disorientation (in 42 percent). Thirteen percent had aseptic meningitis. Hyponatremia developed in 21 percent, and there were signs of increased intracranial pressure in 13 percent. Six patients, including three with cerebral herniation, underwent intracranial-pressure monitoring. The 13 patients (11 percent) whose condition deteriorated in the hospital had decreases in serum sodium levels (P=0.007), and increases in body temperature (P=0.003) at the time of deterioration. At admission, these patients more often had a history of vomiting (P=0.047) and a score of 12 or lower on the Glasgow Coma Scale (P=0.02) than the others; a trend toward a greater prevalence of seizures at admission was also evident in this group (P=0.07). All the patients survived, but 15 of them (12 percent) had neurologic deficits at discharge. Follow-up assessments, performed in 28 children, suggested an increase in cognitive and behavioral deficits 10 to 18 months after the episode of encephalitis. CONCLUSIONS: La Crosse virus infection should be considered in children who present with aseptic meningitis or encephalitis. Hyponatremia and increasing body temperature may be related to clinical deterioration.     

儿童病毒性脑炎270例脑电图与脑电地形图分析

目的探讨脑电图（EEG）及脑电地形图（BEAM）在儿童病毒性脑炎的诊治中的应用价值。方法回顾性分析270例儿童病毒性脑炎的脑电图及脑电地形图的结果，并和脑脊液、CT等的检查结果对比。结果首次EEG异常率为95．18％，BEAM的表现与之相符。治疗后多次复查EEG及BEAM，均恢复正常。结论EEG及BEAM在儿童病毒性脑炎的早期发现异常、提供诊断依据、判断病情严重程度和评估疗效及预后等方面有着其它检测手段所不可取代的作用。     

乙肝病毒基因组中U5序列在乙肝患者垂直传递中的分析研究

目的研究乙肝病毒在乙肝患者垂直传递中的情况，方法运用分子遗传学的技术方法：单核苷酸多态性（SNP）及多聚酶链反应-单链构相多态性（PCR-SSCP），对30个家庭（68个病例）的乙肝患者及其子女进行了研究。结果在乙肝患者及其受感染后出生子女中游离型及整合型乙肝病毒持续增高，与感染前所生子女间比较差异显著（P〈0．05），SNP分析结果显示在乙肝病毒基因组中，U5序列和非U5序列中许多基因位点发生了碱基替换，插入或缺失，男性乙肝患者及其受感染子女的SNP分析结果被确定。结论乙肝病毒可通过男性患者传递至子女；至此，又一个分子遗传学证据证实了了乙肝病毒的遗传性传递。     

Factor analysis reveals differences in brain metabolism in macaques with SIV/AIDS and those with SIV-induced encephalitis

MRS has often been used to study metabolic processes in the HIV-infected brain. However, it remains unclear how changes in individual metabolites are related to one another in this context of virus-induced central nervous system dysfunction. We used factor analysis (FA) to identify patterns of metabolite distributions from an MRS study of healthy macaques and those infected with simian immunodeficiency virus (SIV) which were moribund with AIDS. FA summarized the correlations from nine metabolites into three main factors. Factor 3 identified patterns that discern healthy animals from those with SIV/AIDS. Factor 2 was able to differentiate between animals that had encephalitis and those moribund with AIDS but lacking encephalitis. Specifically, Factor 2 was able to distinguish animals with moderate to severe encephalitis from animals with mild or no encephalitis as well as uninfected controls. FA not only confirmed the involvement of neuronal metabolites (N-acetylaspartate and glutamate) in disease severity, but also detected changes in creatine and myo-inositol that have not been observed in the SIV macaque model previously. These results suggest that the divergent pathways of N-acetylaspartate and creatine in this disease may enable the commonly reported ratio N-acetylaspartate/creatine to be a more sensitive marker of disease severity. Copyright (c) 2008 John Wiley & Sons, Ltd.     

Breastfeeding Might Have Protective Effects on Atopy in Children With the CD14C-159T CT/CC Genotype

Breastfeeding is widely recommended to reduce risk of sensitization, eczema and asthma. However, the role of breastfeeding in prevention of allergic diseases is uncertain. We aimed to investigate whether the relationship between breastfeeding and sensitization to aeroallergens is modified by cluster of differentiation 14 (CD14) genotype. This study included 1,828 school children aged 9-12. We administered a detailed questionnaire and genotyped the CD14C-159T polymorphism. Skin prick tests for 12 aeroallergens were performed. School children who had been breastfed were less likely sensitized to aeroallergens (adjusted odds ratio [aOR] 0.712, 95% confidence interval [CI]: 0.555-0.914). There was no significant association between CD14C-159T genotype and atopy. Breastfeeding was associated with a decreased risk of atopic sensitization in children with CT/CC genotype (aOR 0.667, 95% CI: 0.463-0.960). Our data might identify the gene-environment interaction between the CD14C-159T polymorphism and breastfeeding in relation to aeroallergen sensitization.     

Lack of association between atopic eczema/dermatitis syndrome and polymorphisms in the promoter region of RANTES and regulatory region of MCP-1

BACKGROUND: Chemokines play an important role in the pathophysiology of atopic eczema/dermatitis syndrome (AEDS) and allergy. Recently polymorphisms in the promoter region of RANTES (regulated on activation normal T cell expressed and secreted) and in the gene regulatory region of MCP-1 (monocyte chemoattractant protein-1) have been found, which increase the expression of these chemokines. The - 403A allele of the RANTES promoter region was found associated with AEDS in German children. We investigated whether the presence of these polymorphisms was associated with AEDS or allergy in Hungarian children. METHODS: One hundred and twenty-eight children with AEDS, 102 allergic children without AEDS and 303 children of comparable ages without allergic disorders were screened for genotype with a PCR-based assay. RESULTS: There were no significant differences in the frequency of these polymorphisms, or in the distribution of genotypes between the groups. The total IgE concentration, the white blood cell count and the blood eosinophil cell count did not differ between the genotypes. CONCLUSION: In this cohort of Hungarian children there was no association between - 28G, and - 403A alleles in the RANTES promoter, - 2518G polymorphism in the distal regulatory region of the MCP-1 and AEDS, or allergy.