pp65抗原血症检测法诊断婴儿巨细胞病毒感染的研究

目的　探讨pp65抗原血症检测方法（AA）对新生儿和小婴儿巨细胞病毒（CMV）感染的诊断价值。方法　对21例有典型CMV感染症状患儿及42例无典型症状的患儿,进行外周血AA、尿PCR及血清ELISA法IgM检测,比较3种方法的检测结果,结合抗原血症的量化结果及对部分患儿的短期随访,分析AA的临床应用价值。结果　（1）63例中CMV症状性感染15例,亚临床型感染8例。症状性感染组AA、PCR、ELISA阳性检出率分别为93%（14/15）、100%（15/15）和80%（12/15）;亚临床型感染组阳性检出率分别为50%（4/8）,75%（6/8）和38%（3/8）。AA相对于PCR的敏感性为86%(18/21),特异性为100%。（2）症状性感染通常伴高水平抗原血症［(4～206)个/1.5×105多形核白细胞（PMNL）］,亚临床型感染通常抗原血症水平较低［（0～3）个/（1.5×105）PMNL］,症状性感染组抗原血症水平明显高于亚临床型感染组（P＜0.01）。（3）症状性感染组中4例婴儿肝炎患儿恢复期AA转为阴性或低水平。结论　pp65抗原血症检测方法是一种敏感、特异的检测方法,量化结果有助于诊断活动感染及监测病情。     

人类巨细胞病毒感染的抗原血症检测方法

人类巨细胞病毒 ( Humancytomegalovirus,HCMV)是引起先天和围产期感染 ,导致先天畸形、智力低下、发育迟缓等后遗症的最常见病原 [1 ,2 ]。先天 HCMV感染的发病率为 0 .5 %～ 2 .5 % ,国外报道 :有症状者病死率 30 % ,存活者后遗症发生率70 % ,无症状患儿 5 %～ 1 5 %生长发育中出现异常 [1 ] ,早期、快速、准确的诊断 HCMV活动感染是近年儿科尤其是新生儿科医师一直关心的问题 ,然而目前仍缺少鉴别潜伏和活动感染的有效手段。病毒活动感染时 ,受累的外周白细胞表达病毒抗原 ,用单克隆抗体通过免疫染色检测病毒抗原的方法称为抗原血…     

流式细胞术检测PP65抗原血症在诊断巨细胞病毒感染上的应用

目的　探讨流式细胞技术检测PP6 5抗原血症对巨细胞病毒 (CMV)感染的诊断价值。方法　对 136例CMV感染患儿流式细胞技术检测血PP6 5抗原、实时荧光定量PCR(RFQ PCR)检测尿CMVDNA及血清ELISA法检测CMVIgM ,比较 3种方法的检测结果 ,并以 35例非CMV感染的患儿作为PP6 5检测的阴性对照。对 18例PP6 5抗原阳性患儿进行短期随访。结果　 136例CMV感染患儿从血单个核细胞 (MNC)中检出PP6 5抗原 118例 (118/ 136 ,阳性率 86 8% )。其中 5 3例患儿分别作MNC和多形核白细胞 (PMNL)检测 ,从MNC中检出PP6 5抗原 4 5例 (45 / 5 3,阳性率 84 9% ) ,PMNL中检出PP6 5抗原 4 3例 (43/ 5 3,阳性率 81 1% ) ,MNC与PMNL阳性率差异无统计学意义 (χ2 m=0 2 5 ,P >0 0 5 )。PP6 5与尿CMVDNA2种方法所检测的结果间差异无统计学意义 (χ2 m=1 78,P >0 0 5 ) ;但PP6 5与血CMVIgM检测结果间差异有统计学意义 (χ2 m=5 2 92 ,P <0 0 1)。PP6 5抗原水平与CMV感染的病情程度有明显相关性 ,全身性感染通常伴高水平抗原血症 ,单脏器感染抗原水平相对较低或阴性(χ2 =38 5 1,P <0 0 0 5 )。PP6 5在感染恢复期转为阴性或低水平 ,病情恶化时明显增高。结论　流式细胞技术检测PP6 5抗原血症是诊断活动性CMV感染的有效手段 ,量化结     

pp65抗原与人巨细胞病毒感染

人巨细胞病毒(human cytomegalovirus,HCMV)感染可引起肝脏疾病、肺炎、腹泻病、血小板减少性紫癜、听力障碍等疾病,具有较高的感染率与病死率.pp65(phosphoprotein 65)作为病毒颗粒的主要结构蛋白,与病毒基因表达、宿主免疫逃避及细胞代谢相关.pp65检测目前被认为诊断活动性巨细胞病毒感染的"金标准"之一,且其抗原水平与HCMV感染的临床症状呈正相关性,可用于指导临床"抢先治疗"并改善预后.该文就pp65的蛋白特性、致病机制、诊断方法及其在诊断和预防的应用等方面作一综述.     

新生儿高胆红素血症与人巨细胞病毒感染的关系

为了解高胆红素血症（简称高胆）新生儿中人巨细胞病毒（ＨＣＭＶ）感染状况。方法：用聚合酶链反应（ＰＣＲ）技术对１２０例１２～２８天高胆新生儿的血、尿进行ＨＣＭＶ－ＤＮＡ序列测定。结果：因出生有窒息史、感染致高胆的新生儿中ＨＣＭＶ－ＤＮＡ阳性率分别为３．１％（１／３２）,８．１％（３／３７）。因高胆入院而找不到任何原因的数生儿占１２０例高胆新生儿的１９．２％,与高胆低出生体重儿的ＨＣＭＹ－ＤＮＡ阳性率     

新生儿高胆红素血症与人巨细胞病毒感染的关系

为了解高胆红素血症(简称高胆)新生儿中人巨细胞病毒(HCMV)感染状况.方法用聚合酶链反应(PCR)技术对120例12～28天高胆新生儿的血、尿进行HCMV-DNA序列测定.结果因出生有窒息史、感染致高胆的新生儿中HCMV-DNA阳性率分别为3.1%(1/32),8.1%(3/37).因高胆入院而找不到任何原因的新生儿占120例高胆新生儿的19.2%,与高胆低出生体重儿的HCMV-DNA阳性率分别为43.5%,36.8%.结论部分未明原因的高胆新生儿和高胆低出生体重儿为HCMV感染者.     

病毒抗原血症检测方法诊断活动性HCMV感染

人类巨细胞病毒（ＨＣＭＶ）常在免疫缺陷或免疫受抑制的病人中引起活动性感染，表现为多种临床疾病，具有较高的病死率。ＨＣＭＶ抗原血症检测方法具有准确性高、技术简便、可快速诊断并人有定量分析的特点，是目前临床诊断活动性ＨＣＭＶ感染的一种重要方法。本文综述了对外周血白细胞应用单克隆抗体技术和免疫染色方法进行ＨＣＭＶ抗原检测以诊断活动性ＨＣＭＶ感染的方法和可能的质量监控手段。     

早期诊断和监测系统性红斑狼疮患儿活动性人类巨细胞病毒感染的研究

目的：了解系统性红斑狼疮（systemic lupus erythematosus,SLE)患儿活动性人类巨细胞病毒（human cytomegalovirus,HCMV)感染的状况,并比较不同实验室检测方法的诊断价值,方法：实验组观察了２１例初诊为ＳＬＥ并接受免疫抑制治疗的患儿,对照组观察了２１例免疫力正常的骨科患儿,治疗前后应用间接免疫荧光法检测外周自多形核白细胞（polymorphonuclear leukocytes,PMNLs）中的ＨＣＭＶpp65抗原和p７２抗原,ＰＣＲ方法检测血清中的ＨＣＭＶＤＮＡ,ＥＬＩＳＡ方法检测血清中的ＨＣＭＶＩgM和ＩgG抗体,结果,实验组活动性ＨＣＭＶ感染发生率为２９％（６／２１）,４例发生于免疫抑制治疗后,另外２例发生免疫抑制治疗前,对照组无１例发生活动性感染,两组相比差异有显著性（Ｐ＝０．０２７）,实验组各项实验室检测的阳性率分别为:pp6524%(5/21),p7214%(3/21),DNA43%(9/21),IgM10%(2/21),IgG91%(19/21).实验组各项实验室检测方法诊断活动性ＨＣＭＶ感染的敏感性分别为：pp6583%(5/6),p7250%(3/6),NDA100%(6/6),IgM33%(2/6),IgG(双份血清抗体滴度呈≥４倍增高）５０％（３／６）,特异性ＰＣＲ方法为８０％（１２／１５）外,其余均为１００％（１５／１５）,结论：ＳＬＥ患儿活动性ＨＣＭＶ感染发生率较免疫力正常的儿童显著增高,免疫抑制治疗前后可发生,ＨＣＭＶpp65抗原血症检测是早期诊断和监测ＳＬＥ患儿活动性ＨＣＭＶ感染的较好指标。     

血尿患儿尿和肾组织中巨细胞病毒抗原检测及其临床意义

目的 探讨血尿患儿巨细胞病毒（CMV）感染状况。方法 应用免疫组化链霉菌抗生物素蛋白－地氧化物酶连结法,检测55例血抗CMV－IgM阳性的血尿患儿尿中巨细胞病毒早期抗原（CMV－EA）,并对其中6例行肾穿刺活检,进行肾组织中CMV－EA检测。结果 单纯性血尿组患儿尿CMV－EA阳性率为80％（32／40）。非单纯性血尿组患儿尿CMV－EA阳性率为20％（3／15）。对照组尿CMV－EA阳性率为19％（10／53）。单纯性血尿组尿MCV－EA阳性率明显高于非单纯性血尿组和对照组（P＜0．005,P＜0．05）。6例肾组织CMV－EA检测5例阳性（83％）。肾组织CMV感染细胞多为皮质区小管上皮细胞。结论 巨细胞病毒感染可能是单纯性血尿的主要病因。     

Clinical importance of cytomegalovirus antigenemia for intrauterine cytomegalovirus infection

Background:  Little is known about the clinical importance of cytomegalovirus (CMV) antigenemia for intrauterine-CMV-infected newborns. The aims of the present study were to evaluate the diagnostic accuracy of CMV antigenemia during the neonatal period and its association with clinical manifestations.
Methods:  CMV antigenemia was analyzed using neonatal blood from 25 patients suspected of having intrauterine infection because of abnormal clinical manifestations in the mother, fetus, and newborn. Neonatal urine samples were collected for diagnosis of intrauterine infection. The diagnostic accuracy of the antigenemia analysis was evaluated by comparing it with the results of urinary CMV analyses. The clinical manifestations of antigenemia-positive and -negative infected newborns were compared in the infected newborns.
Results:  Fifteen newborns were congenitally infected and 10 were uninfected as diagnosed on virus isolation from neonatal urine. Six of 15 infected newborns were positive for CMV antigenemia. CMV antigenemia had a positive predictive value of 100%, a negative predictive value of 52.6%, a sensitivity of 40%, and a specificity of 100%. CMV retinitis and pneumonitis were more prevalent among antigenemia-positive newborns (4/6) than antigenemia-negative newborns (0/9; P  < 0.05).
Conclusions:  Antigenemia was significantly associated with retinitis and pneumonia, but it was not sensitive enough to diagnose intrauterine CMV infection.     

pp65抗原血症检测法监测婴儿巨细胞病毒肝炎的研究

目的探讨pp65抗原血症检测方法(AA)对婴儿巨细胞病毒(HCMV)肝炎临床应用价值。方法对2001年11月～2002年10月入院的76例婴儿肝炎患儿应用间接荧光染色法检测外周血pp65抗原血症,并与血清ELISA法检测HCMVIgM进行比较。对部分婴儿HCMV肝炎患儿应用更昔洛韦治疗后短期随访。结果76例中64例为HCMV肝炎。AA、ELISA法HCMV阳性检出率分别为92.2%(59/64例)、64.1%(41/64例)。抗原血症呈高水平、中水平者所占比例分别为81.4%(48/59例)、18.8%(11/59例),无低水平者。47例婴儿HCMV肝炎更昔洛韦治疗2周,治疗前、后抗原血症水平,经卡方检验,P<0.01。治疗后随访中肝功能正常与肝功能异常组,抗原血症水平经精确概率法检验,P<0.01。结论pp65抗原血症检测方法有助于监测婴儿巨细胞病毒肝炎的动态变化,指导治疗。     

小儿人类免疫缺陷病毒感染的临床诊断和处置原则

艾滋病即获得性免疫缺陷综合征(acquiredimmunode ficiencysyndrome ,AIDS) ,是由人类免疫缺陷病毒(HIV)感染所致的一种传播迅速、病死率极高的恶性传染病。小儿HIV感染主要由母婴传播途径获得,其次由输入的血液(全血和血浆)和血液制品获得。小儿HIV感染发生率的增长较成人快、潜伏期短,疾病进展快、病死率高。因此小儿HIV感染暨艾滋病防治已是我国儿科界所面临的严峻挑战和紧迫任务。1　HIV感染和AIDS的诊断小儿HIV感染和AIDS的诊断,需结合流行病史、临床和实验室检查等进行综合分析。小儿HIV感染包括无症状HIV感染和AIDS…     

婴儿巨细胞病毒感染的诊治

巨细胞病毒 (ｃｙｔｏｍｅｇａｌｏｖｉｒｕｓ,ＣＭＶ )为双链ＤＮＡ病毒 ,属于疱疹病毒科 ,与其它疱疹病毒一样 ,急性感染恢复后 ,病毒可长期潜伏在体内 ,一旦机体免疫功能减损 ,病毒即可激活致病 ,故为“机会”致病微生物。ＣＭＶ感染可引起多种不同的感染综合征 ;在新生儿可引起先天性ＣＭＶ感染 ,在正常健康人可引起单核细胞增多症 ;在免疫缺陷患者 ,如未成熟新生儿、器官移植受者或艾滋病 (ＡＩＤＳ)患者等 ,ＣＭＶ可引起严重的疾病综合征。1　婴儿ＣＭＶ感染的流行病学特点ＣＭＶ感染是宫内感染中最常见的病原 ,先天性ＣＭＶ感染约占…     

Ganciclovir therapy for congenital cytomegalovirus infection in six infants

BACKGROUND: Congenital cytomegalovirus (CMV) infection is common, and its morbidity rate is high. Ganciclovir (GCV) treatment has been used for congenital CMV infection, but there are few reports on viral loads associated with GCV therapy. METHODS: A real-time PCR assay was used to monitor viral load in 6 cases of symptomatic CMV infection that received GCV therapy. Initially GCV was given at a dose of 5-12 mg/kg/d for 2-7 weeks. In 2 cases, additional doses were given as symptoms returned. RESULTS: After GCV administration, active signs of chorioretinitis, thrombocytopenia and anemia disappeared or improved in all cases. During GCV therapy, viral loads decreased while patients improved clinically and increased again when GCV therapy was stopped. Although CMV DNA continued to be detectable for a long period, clinical findings did not always worsen. In 2 cases, an improvement of hearing loss was observed. CONCLUSION: GCV therapy transiently suppresses the CMV concentrations. Subsequent increases of viral titers do not appear to be correlated with the clinical course or neurologic outcome.     

Breast milk and cytomegalovirus infection in preterm infants

The incidence of postnatal human cytomegalovirus (HCMV) reactivation during lactation equals the maternal seroprevalence. Infectious virus, viral DNA and RNA can be isolated from cells and fat free milk whey. Early onset of viral DNAlactia and virolactia as well as high viral load in milk whey are maternal risk factors for virus transmission. Preterm infants below 1000 g birthweight and a gestational age below 30 weeks may be at high risk of acquiring a symptomatic HCMV infection. Several recent studies using frozen milk for feeding describe low transmission rates and mostly asymptomatically infected neonates. However despite different freeze-storing procedures HCMV transmission occurred and severe HCMV infections were observed. Only few data exist on the long-term outcome of postnatally acquired HCMV infection via breast milk. Additional long-term outcome studies are needed. The newly developed short-term pasteurisation may be a reliable alternative to freezing and Holder pasteurisation, since important milk constituents are conserved.     

Clinical applications of real-time PCR for diagnosis and treatment of human cytomegalovirus infection in children

There are many methods of detecting human cytomegalovirus (HCMV) infection. So far, the quantitative polymerase chain reaction (PCR) has been very useful not only in aiding in the diagnosis of HCMV but also in determining the severity and predicting HCMV infection. However, it is time-consuming and labor intensive. Real-time PCR (RT-PCR) is an exception, for it allows rapid quantification of HCMV DNA load. Our group used this method for detecting and monitoring HCMV and compared it with the diagnostic criterion recommended by the Pediatric Branch of Chinese Medical Association, in 45 children suspected of having HCMV infection. The response to two types of antiviral treatment on HCMV DNA load was also monitored in HCMV hepatitis cases. RT-PCR was positive in 30 cases while the diagnostic criterion, which includes enzyme-linked immunosorbent assay (ELISA) and/or conventional PCR, was positive in 32 cases. The decrease in the HCMV DNA load was achieved earlier in the modified treatment group compared with the conventional treatment group. A 10(3) copies/ml of HCMV DNA load of is a useful cut-off value in predicting patients who will have symptoms of the disease. RT-PCR can be used not only in detecting HCMV but also in monitoring response to antiviral treatment and risk of having symptoms of the disease.