逆转录多聚酶链反应对肠道病毒性脑膜炎、脑炎诊断价值探讨

目的探讨中枢神经系统感染的病原体及其检测方法。方法应用逆转录多聚酶链反应（ＲＴ－ＰＣＲ）检测６４例无菌性脑膜炎及２７例无菌性脑炎患儿脑脊液（ＣＳＦ）中肠道病毒（ＥＶ）ＲＮＡ。结果急性期ＣＳＦ中ＥＶＲＮＡ阳性检出率为：无菌性脑膜炎６０９％,脑炎４８１％;阳性患儿恢复期ＣＳＦ仅个别阳性;对照组全部阴性。起病５天以内阳性率明显高于５天以后（Ｐ＜００５）;５～１０月份较１１～４月份发病率高（Ｐ＜００５）。２５例病毒分离阳性者中２４例ＲＴ－ＰＣＲ阳性,５３例病毒分离阴性者中１９例ＲＴ－ＰＣＲ阳性,ＲＴ－ＰＣＲ敏感性显著高于病毒分离的敏感性（Ｐ＜００１）。结论ＥＶ是引起中枢神经系统感染的重要病原体,ＲＴ－ＰＣＲ技术是诊断ＥＶ感染的有效方法。     

逆转录聚合酶链反应快速检测呼吸道合胞病毒

为探索一种快速、可靠的呼吸道合胞病毒（ＲＳＶ）的检测方法，对逆转录聚合酶链反应（ＲＴ－ＰＣＲ）技术检测ＲＳＶ进行研究。结果：ＲＴ－ＰＣＲ方法可检出病毒量为１０ＴＣＩＤ５０／ｍｌ（ＴＣＩＤ５０为半数组织培养感染量）；用于扩增Ｆ蛋白基因的引物经计算机基因库检测与其他病毒无同源性，实验同步扩增其他常见呼吸道感染病毒均无阳性扩增带；实验过程６小时内完成。在ＲＳＶ好发季节对５３例拟诊为病毒性急性下呼吸道感染婴儿咽拭子进行检测，ＲＳＶ阳性２８例（５２．８％）。提示ＲＴ－ＰＣＲ方法检测ＲＳＶ具有快速、敏感性高、特异性好等特点，适合于临床应用。     

逆转录—多聚酶链反应检测柯萨奇B组病毒

研究目的用分子生物学方法对柯萨奇Ｂ组病毒（ＣＢＶ）进行检测并探讨与临床症病的关系。研究方法对我院２９８例疑似ＣＢＶ感染的患儿，采用特异性引物进行逆转录－多聚酶链反应（ＲＴ－ＰＣＲ）检测ＣＢＶ。结果检出ＣＢＶ阳性１２７人，阳性率４２．６％。感染与年龄有关，夏秋季多见。８０例健康儿童中，阳性５人，占６．２５％。２０例ＲＴ－ＰＣＲ检测阳性标本用ＥＬＩＳＡ法检测，１７例阳性。结论ＲＴ－ＰＣＲ可快速、敏感、准确地检出标本中的ＣＢＶ。此病毒可侵犯多种器官，引起各种临床症病，有较强的传染性。     

非化脓性中枢感染中肠道病毒感染的临床研究

目的 分析１９９７年度非化脓性中枢感染中肠道病毒（ＥＶ）感染的临床特点。方法 采用ＲＴ－ＰＣＲ的方法检测１９９７年１２８例非化脓性中枢感染者的ＣＳＦ中ＥＶ ＲＮＡ。结果与我院非化脓性中枢感染季节分布情况比较。结果 本组年龄与ＥＶ感染关系不大，起病早期ＣＳＦ阳性检出高，非化脓性中枢感染与ＥＶ感染关系不大，起病早期ＣＳＦ阳性检出高，非化脓性中枢感染与ＥＶ在ＣＳＦ中检出一致。结论 ＥＶ是小儿时期非化脓性     

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．     

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     

新生儿室管膜下囊肿的病因研究

为了探讨新生儿室管膜下囊肿（ＳＥＣ）与先天性感染的关系，采用ＥＬＩＳＡ酶标法检测母婴双方囊肿组与对照组各７０例血巨细胞病毒（ＣＭＶ）、风疹病毒、弓形虫的抗体，并应用聚合酶链反应（ＰＣＲ）技术直接检测病原ＤＮＡ（风疹病毒除外），同时作尿ＣＭＶ的ＰＣＲ检测。结果：囊肿组新生儿血ＣＭＶ－ＩｇＭ抗体和ＣＭＶ－ＰＣＲ阳性率显著高于对照组（分别为１７．１％、５．７％和１２．９％、２．９％）；且尿ＣＭＶ－ＰＣＲ的阳性率达４０．０％（２８／７０），高于对照组的１７．１％（１２／７０）；分别进行两组自身对照，尿ＣＭＶ－ＰＣＲ的阳性率显著高于血ＣＭＶ－ＰＣＲ阳性率（Ｐ＜０．０５）。囊肿组母亲的尿ＣＭＶ－ＰＣＲ阳性率亦显著高于对照组（３０％、１０％、Ｐ＜０．０１）。提示新生儿ＳＥＣ是宫内感染损害中枢神经系统的表现之一，与先天性ＣＭＶ感染有关，尿ＣＭＶ－ＰＣＲ检查可作为宫内ＣＭＶ感染的首选实验室诊断方法。     

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     

肠道病毒感染与儿童Ⅰ型糖尿病关系的研究

目的 探讨儿童Ⅰ型糖尿病与肠道病毒（ＥＶ）感染,特别是柯萨奇Ｂ组病毒（ＣＶＢ）感染的关系。方法 采用ＲＴ－ＰＣＲ方法,对４５例糖尿病患儿血中ＥＶ－ＲＮＡ进行检测,用微量中和试验方法,对糖尿病患儿正常小儿血清中抗ＣＶＢ中和抗体水平进行测定,同时用ＥＬＩＳＡ方法,测定ＣＶＢ特异性ＩｇＭ。结果（１）糖尿病线４５例患和中ＥＶ－ＲＮＡ阳性者为１７例,（３９％）,对照组５１名儿童中了性者为３名（６％）。（２）     

巨细胞包涵体病的临床及早期诊断

本文观察１９９２年９月至１９９４年３月我科收治的人类巨细胞病毒（ＨＣＭＶ）感染患儿３４例。部分阳性患儿同时检测母亲ＰＣＲ－ＨＣＭＶ－ＤＮＡ阳性率为８０％，说明患儿ＨＣＭＶ感染与母亲有明显关系。本文采用了聚合链式反应（ＰＣＲ）检测患儿血ＨＣＭＶ－ＤＮＡ的方法，并与血清免疫学方法比较，结果表明，该方法具有高度的特异性和敏感性，操作简便，反应周期短，不存在交叉反应，是目前临床检测ＨＣＭＶ感染较好的方法。     

Hand-foot-and-mouth disease in calicut

Objective: Hand-Foot-and-Mouth Disease (HFMD) is a mild exanthematous illness seen worldwide, affecting mainly children under ten years of age. The causative agents were initially Coxsackie virus type A 16 and related serotypes. The situation changed drastically about thirty years ago with the advent of a new aetiological agent, Enterovirus type 71 (EV 71), which has caused very large outbreaks with severe complications and many deaths.Methods: The authors report an outbreak of papulovesicular lesions on the skin and oral mucosa compatible with the diagnosis of HFMD in children in and around Calicut in October 2003. Clinical and laboratory study in collaboration with the National Institute of Communicable Diseases, Delhi. Eighty one children with the syndrome were examined and followed up from October 2003 to February 2004, when the outbreak subsided.Result: The outbreak was mild and all children recovered within 1 to 2 weeks.Conclusion: Acute and convalescent paired serum samples collected from 19 patients were examined at the NICD for IgM antibody against EV 71 by microneutralisation test in cell culture. All the paired samples tested showed significant rise in titre of antibodies, confirming the diagnosis of EV 71 infection in each of them.     

Nonpolio enterovirus infection in the neonate and young infant

Nonpolio enterovirus infections are common in the neonatal period, accounting for a large portion of febrile illness during the summer months. Unlike older children and adults, some neonates with enterovirus infection progress to multisystem disease and death. Multiple clinical syndromes of varying severity are associated with neonatal enterovirus infection: asymptomatic viral shedding, nonspecific febrile illness, aseptic meningitis, hepatic necrosis and coagulopathy, and myocarditis. In the present paper, a typical case of neonatal febrile illness is presented and the English-language literature is reviewed with respect to enteroviral infection in early infancy. The virology, epidemiology, transmission, clinical features, diagnosis and treatment of neonatal enteroviral infection are presented. Although the majority of infections in the neonate are benign, timely diagnosis in the febrile neonate will expedite efficient management. Clinicians also need to recognize the clinical manifestations and risk factors for severe disease to anticipate complications and implement intensive management of infants at high risk of adverse outcomes.     

无菌性发热新生儿单个核细胞肠道病毒基因表达的研究

目的　探讨无菌性发热新生儿外周血单个核细胞 (PBMC)中肠道病毒 (EV)基因表达情况和临床表现特点。方法　采用逆转录聚合酶链反应 (RT PCR)、原位杂交和病毒分离技术检测了 2 9例发热待查组和 18例中枢感染组新生儿PBMC、脑脊液及血清中EVRNA ,并对其临床特点进行总结分析。结果　发热待查组EV感染 14例 (48 3% ) ,9例PBMC中EVRNA阳性者 ,血清中全部阳性 ,其中 4例脑脊液中同时阳性 ;中枢感染组EV感染 11例 (6 1 1% ) ,5例外周血PBMC中EVRNA阳性者 ,血清和脑脊液中也全部阳性。两组比较 ,只有RT PCR法检测中枢感染组脑脊液中EVRNA阳性数高于发热组 ,而不同方法检测PBMC和血清中EVRNA两组间差异均无显著性。EV感染的主要症状有喂养困难 (71% )、易激惹 (6 4 % )、腹泻 (6 1% )、嗜睡 (5 4 % )、肝脏肿大 (46 % )及皮疹(36 % )等。结论　EV可感染新生儿的PBMC ;新生儿感染后EV易在全身播散 ;临床表现不典型 ,早期、快速病原诊断可缩短住院天数和减少抗生素的应用 ,有较高的社会和经济效益。     

Non-polio enteroviruses in acute flaccid paralysis

Objective Human enteroviruses are the major cause of aseptic meningitis and also cause a wide range of other acute illnesses, including neonatal sepsis like disease, meningitis, acute flaccid paralysis and acute hemorrhagic conjunctivitis. Infection in neonates is particularly life threatening.Methods : Stool samples of 523 children (age < 4 years) showing symptoms of acute flaccid paralysis (AFP) were studied. National Polio Surveillance Project workers from different parts of Uttar Pradesh and Bihar collected the samples during June to October 1998. Non-polio enteroviruses (NPEV) were isolated in 191 cases only, by cell culture based neutralization assay. These NPEV isolates were further studied to find the frequent enterovirus serotype detected in stool of children having AFP.Result : Data generated will help future studies on NPEV serotypes circulating in this area.Conclusion : In addition it may reduce unnecessary hospitalization, allow immune globulin batches of high titres to frequently circulating serotypes, to be reserved for intravenous therapy of neonates and guide the formulation of antigens for rapid and less expensive diagnosis.     

肠道病毒71型感染患儿胸X线片表现

目的：研究肠道病毒71型感染患儿的胸部X线影像特点。方法：选取2010年4月至2011年7月肠道病毒71型感染患儿120例,按病情分成轻型组（31例）、重型组（43例）、危重型组（46例）,对患儿发病至首次拍片时间以及首次胸部X线片影像进行比较。结果：各组发病至首次胸部X线片检查时间分别为：轻型组26~48 h（中位时间37 h）;重型组10~36 h（中位时间23 h）;危重型组2~36 h（中位时间19 h）。最早发现胸部X线片异常的时间轻型组约为发病后30 h;重型组约为发病后23 h;危重型组约为发病后2 h;3组胸部X线片最早出现异常的时间差异有统计学意义（P<0.01）。首次影像异常率：轻型组5.8%,重型组81.3%,危重型组100%,危重型组首次X线片的异常比例明显高于其他两组（P<0.01）。在胸部X线片的表现上,轻型组最常见的征象为肺纹理增粗、模糊;重型组常见表现为渗出、实变影;危重型则表现为肺水肿征像。危重型组病灶分布广,累及多个肺叶。结论：肠道病毒71型感染患儿首次胸片检查时间、胸片出现异常的时间、异常率、胸部X线片表现严重度与临床病情严重程度相关。     

Successful Heart Transplantation Following Neonatal Necrotic Enterovirus Myocarditis

Enterovirus myocarditis is a potentially devastating diagnosis in the neonatal setting, with an associated high mortality and risk for chronically impaired cardiac function. Transplantation may be a possible therapeutic option, but, due to the comparative scarcity of the presentation, the limited donor pool, and fears of viral persistence and recurrence of myocarditis (especially in the immunocompromised, post-transplant setting), there are no reported cases in the literature. This case report illustrates an encouraging one-year outcome following heart transplantation for necrotic enterovirus myocarditis in a neonate.     

小儿肠套叠与肠道病毒感染

采用逆转录聚合酶链式反应分别对原发性肠套叠患儿和健康小儿的粪便标本进行肠道病毒核糖核酸（ＲＮＡ）检测，结果４岁以上年龄组两者肠道病毒阳性率差异无显著性意义（Ｐ＞０．０５），４岁以下年龄组原发性肠套叠肠道病毒阳性率３７．０４％，显著高于健康小儿阳性率１１．１１％（Ｐ＜０．０１），提示肠道病毒感染可能是部分原发性肠套叠的病因之一。