Viremia in neonatal herpes simplex virus infections.

BACKGROUND: Polymerase chain reaction assays of the peripheral blood mononuclear cells (PBMC) and plasma may facilitate the diagnosis of neonatal herpes simplex virus (HSV). METHODS: Assays for HSV DNA were submitted from at least 1 specimen site (PBMC, plasma or cerebrospinal fluid) in 11 consecutive cases of neonatal HSV infection. RESULTS: HSV DNA was detected by PCR in the PBMC of 6 of 10 infants tested (60%), the plasma of 4 of 6 tested (67%) and the cerebrospinal fluid of 4 of 11 tested (36%). CONCLUSIONS: HSV viremia is more frequent than previously appreciated, and detection of HSV DNA in PBMC and plasma is a useful diagnostic tool, particularly in infants without skin lesions.     

快速诊断单纯疱疹病毒脑炎

目的探讨快速诊断单纯疱疹病毒脑炎（ＨＳＥ），比较不同病毒学试验的诊断价值。方法用聚合酶链反应技术检测１７７例急性脑炎患儿的脑脊液（ＣＳＦ）标本中单纯疱疹病毒（ＨＳＶ）特异性ＤＮＡ；用酶联免疫吸附方法检测ＣＳＦ和血清标本中ＨＳＶ特异性ＩｇＭ和ＩｇＧ抗体。结果ＣＳＦ中ＨＳＶ特异性ＤＮＡ、ＩｇＭ和ＩｇＧ抗体阳性率分别为１．７％（３／１７７）、１０％（１／１００）和４７０％（４７／１００），血清ＨＳＶＩｇＭ、ＩｇＧ抗体阳性率分别为１２．５％（６／４８）、７２．９％（５１／７０）（因为标本量不足或缺如，未能对全部病例进行抗体检测）；３例患儿确诊为ＨＳＥ。结论用套式ＰＣＲ检测ＣＳＦ诊断ＨＳＥ较敏感、特异。     

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.     

宏基因组二代测序在伴有中枢神经系统受累儿童噬血细胞综合征中的应用价值

目的 探究宏基因组二代测序(metagenomic next-generation sequencing,mNGS)技术检测伴有中枢神经系统受累的噬血细胞综合征患儿颅内EB病毒(Epstein-Barr virus,EBV)感染的应用价值及临床意义。方法 回顾性分析30例伴有中枢神经系统受累的噬血细胞综合征患儿的脑脊液mNGS结果,与脑脊液EBV-DNA定性检测、血清EBV抗体谱检测结果进行比较,以治疗前后血清EBV-DNA拷贝数变化反映针对性治疗效果。结果 脑脊液mNGS EBV检测阳性率为100%(30/30),高于脑脊液EBV-DNA定性检测阳性率(10%,3/30;P<0.001),与血清EBV抗体谱检测阳性率(93%,28/30)比较差异无统计学意义(P>0.05)。中位mNGS EBV检出序列数为2 400,治疗前血清EBV-DNA拷贝数与EBV检出序列数呈中度正相关(rs=0.693,P<0.001)。多元线性回归分析结果显示,治疗前血清EBV-DNA拷贝数越高,脑脊液mNGS EBV检出序列数越高(P<0.05)。结论 EBV相关噬血细胞综合征容...     

Determination of the six major human herpesviruses in cerebrospinal fluid and blood specimens of children

AIM: To detect and differentiate six major human herpesviruses in the cerebrospinal fluid (CSF) and blood of children by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). METHODS: We synthesized two pairs of primers in the well-conserved regions of the DNA polymerase gene in human herpesviruses. One pair was designed to amplify cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), and the other pair to amplify varicella-zoster virus (VZV) and human herpesvirus 6 (HHV-6) by PCR. Virus species identification was achieved by restriction enzyme digestion with BamHI and BstUI. Ninety-eight CSF and 75 blood specimens were analysed by this technique. At the same time, all blood specimens were also examined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Thirteen (13.3%) of 98 CSF specimens and 26 (34.7%) of 75 blood specimens were positive for herpesvirus DNA in this PCR assay. Only 10 (13.3%) of the blood specimens were positive in ELISA for virus-IgM antibody. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of PCR in detecting herpesvirus infections compared with ELISA were 100% (10/10), 75.4% (49/65), 38.5% (10/26) and 100% (49/49), respectively. These results indicate that the positive rate of PCR was significantly higher than that of ELISA (p < 0.05). The herpesvirus type of these positive specimens was rapidly detected using restriction enzyme digestion with BamHI and BstUI. CONCLUSIONS: PCR-RFLP is a specific, sensitive and accurate technique for the identification of herpesvirus infections in the CSF and blood of children.     

FilmArray脑炎/脑膜炎多重病原体核酸联检试剂盒在中枢神经系统感染病原学诊断中的价值

目的:探讨FilmArray脑炎/脑膜炎多重病原体核酸联检试剂盒(FilmArray ME Panel)在中国儿童中枢神经系统感染病原学诊断方面的应用价值。方法:收集2019年3月至11月上海交通大学医学院附属上海儿童医学中心收治的145例疑诊中枢神经系统感染患儿的经腰椎穿刺获得的脑脊液标本,对所有患儿的脑脊液标本行培...     

Clinical and laboratory features of neonatal herpes simplex virus infection: a case-control study

BACKGROUND: Neonatal herpes simplex virus (HSV) infection can cause significant morbidity and mortality but can be difficult to identify, particularly in neonates without vesicular rash. OBJECTIVE: To determine the unique clinical and laboratory features of neonates with and without HSV infection admitted to Texas Children's Hospital during a 14-year period. METHODS: An historic case-control study of all hospitalized neonates with laboratory-confirmed HSV infection and a restricted sample (ratio 1:4) of HSV test-negative hospitalized neonates. Univariate and multivariate analyses were performed to identify clinical and laboratory factors associated with neonatal HSV infection. RESULTS: Forty cases and 160 comparison subjects were identified. The following factors were associated with neonatal HSV infection by univariate analysis: maternal primary HSV infection, maternal fever, vaginal delivery, prematurity, postnatal HSV contact, vesicular rash, hypothermia, lethargy, seizures, severe respiratory distress, hepatosplenomegaly, thrombocytopenia, elevated hepatic enzymes, and cerebrospinal fluid (CSF) pleocyosis and proteinosis. Factors not associated with neonatal HSV infection were fever, total peripheral white blood cell count, and red blood cells in the CSF. For neonates presenting without vesicular rash, maternal fever, respiratory distress requiring mechanical ventilation, and CSF pleocytosis were independently associated with HSV infection. CONCLUSIONS: Inclusion of the newly appreciated features of maternal fever, respiratory distress, and thrombocytopenia might improve the detection of neonatal HSV infection. Clinical and laboratory factors typically associated with neonatal HSV infection were confirmed to be maternal primary HSV infection, vaginal delivery, prematurity, neonatal seizures, vesicular rash, elevated hepatic enzymes, and CSF pleocytosis.     

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。     

Persistence of antibody to hepatitis B and protection from disease among Alaska natives immunized at birth

BACKGROUND: Alaska Native (AN) children were at high risk of acquiring hepatitis B virus (HBV) infection before vaccination began in 1983. We evaluated the long-term protection from hepatitis B (HB) vaccination among AN children immunized when infants. METHODS: During 1984-1995, we recruited a convenience sample of AN children who had received a three dose series of HB vaccine starting at birth and had serum antibody to hepatitis B (anti-HBs) concentrations of >/= 10 mIU/mL at 7-26 months of age. We evaluated anti-HBs concentrations and the presence of anti-HBc in participants' sera every other year up to age 16 years. Anti-HB core antigen (anti-HBc)-positive specimens were tested for hepatitis B surface antigen and for HBV DNA. RESULTS: We followed 334 children for 3151 person-years (median, 10 years per child) with 1610 specimens collected. Anti-HBs concentrations dropped rapidly among all participants. Among children 2, 5 and 10 years of age, 37 of 79 (47%), 33 of 176 (19%) and 8 of 95 (8%), respectively, had anti-HBs concentrations of >/= 10 mIU/mL. Receipt of recombinant vaccine was significantly associated with a more rapid antibody decline (P < 0.001). Six (1.8%) children acquired anti-HBc, 3 of whom had definite breakthrough infections (at least 2 consecutive anti-HBc-positive specimens or at least 1 anti-HBc-positive specimen and HBV DNA detection by PCR). None of these children had detectable hepatitis B surface antigen, and none had symptoms of hepatitis. CONCLUSIONS: Anti-HBs concentrations declined over time among AN infants successfully immunized with HB vaccine starting at birth. Transient anti-HBc appeared in a small percentage of children; however, none developed clinical signs of hepatitis or chronic HBV infection.     

Elevated serum levels of tumor necrosis factor are associated with progressive encephalopathy in children with acquired immunodeficiency syndrome

The cytokine tumor necrosis factor (TNF) was assayed in the sera (n = 31) and cerebrospinal fluid (n = 26) of children with acquired immunodeficiency syndrome, using a competitive radioimmunoassay. Elevated serum levels of TNF were found in 15 (79%) of 19 patients with progressive encephalopathy (PE), compared with 1 (8%) of 12 patients without neurologic involvement. There was a significant association of PE with elevated serum TNF levels. Conversely, of 16 patients with elevated serum TNF levels, 15 (94%) were found to have PE, and of 8 patients with serum TNF levels greater than 100 pg/ml, all 8 (100%) had PE. No association was found between cerebrospinal fluid levels of TNF and PE. Neither serum nor cerebrospinal fluid TNF levels correlated with the degree of cachexia. These data suggest that circulating TNF may be responsible for the myelin damage that occurs in human immunodeficiency virus type 1-associated PE.     

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。     

Isolation of measles virus from cerebrospinal fluid of children with acute encephalopathy without rash.

OBJECTIVE: To determine the viral agent involved in cases of acute encephalopathy in children during an outbreak in Northern India. DESIGN: Virological and serological studies using serum and cerebrospinal fluid specimens from patients. METHODS: Serum and CSF specimens were tested by IgM ELISA for IgM antibodies to variety of viruses like Japanese encephalitis, West Nile, Dengue and Measles. The specimens were inoculated into Vero cell monolayer for virus isolation. The viral strains isolated were identified by indirect immunofluorescence test and qualitative in-vitro neutralization test using polyclonal and monoclonal antibodies to measles. Identity of the isolates was reconfirmed using RT-PCR method. RESULTS: Of the 28 specimens tested, 17 had IgM antibodies to measles. Commercial IgM ELISA kits confirmed the serological findings. Vero cell cultures yielded 4 isolates from CSF and 2 from serum specimens of six different patients. Cytopathic effect was typical of measles. Indirect imunofluorescence using polyclonal and monoclonal antibodies to measles HA protein, confirmed the measles etiology. Neutralization tests reconfirmed the measles strain isolation. RT-PCR amplified product was confirmed as measles. CONCLUSION: The isolation of measles virus from CSF and serum of children with acute encephalopathy without rash proved the etiological role of measles virus in this outbreak.     

Suboptimal management of central nervous system infections in children: a multi-centre retrospective study

ABSTRACT: OBJECTIVE: We aimed to audit the regional management of central nervous system (CNS) infection in children. METHODS: The study was undertaken in five district general hospitals and one tertiary paediatric hospital in the Mersey region of the UK. Children admitted to hospital with a suspected CNS infection over a three month period were identified. Children were aged between 4 weeks and 16 years old. Details were recorded from the case notes and electronic records. We measured the appropriateness of management pathways as outlined by national and local guidelines. RESULTS: Sixty-five children were identified with a median age of 6 months (range 1 month to 15 years). Ten had a CNS infection: 4 aseptic meningitis, 3 purulent meningitis, 3 encephalitis [2 with herpes simplex virus (HSV) type 1]. A lumbar puncture (LP) was attempted in 50 (77%) cases but only 43 had cerebrospinal fluid (CSF) available for analysis. Of these 24 (57%) had a complete standard set of tests performed. Fifty eight (89%) received a third generation cephalosporin. Seventeen (26%) also received aciclovir with no obvious indication in 9 (53%). Only 11 (65%) of those receiving aciclovir had CSF herpes virus PCR. Seventeen had cranial imaging and it was the first management step in 14. Treatment lengths of both antibiotics and aciclovir were highly variable: one child with HSV encephalitis was only treated with aciclovir for 7 days. CONCLUSIONS: The clinical management of children with suspected CNS infections across the Mersey region is heterogeneous and often sub-optimal, particularly for the investigation and treatment of viral encephalitis. National guidelines for the management of viral encephalitis are needed.     

Absence of associations between influenza-associated encephalopathy and human herpesvirus 6 or human herpesvirus 7

BACKGROUND: Influenza-associated encephalopathy is a severe complication of influenza virus infection, but its pathogenesis is unknown. It was recently suggested that the neurologic complications of influenza, including encephalopathy, are associated with human herpesvirus 6 (HHV-6) and human herpesvirus 7 (HHV-7).AIM To confirm or refute the association between influenza-associated encephalopathy and HHV-6 or HHV-7. METHODS: Cerebrospinal fluid and serum from 25 patients with central nervous system complications of influenza (18 patients with encephalopathy and 7 patients with febrile convulsions) were investigated. The specimens were examined by real time polymerase chain reaction (PCR) and nested PCR for HHV-6 and HHV-7 DNA. RESULTS: In the cerebrospinal fluid samples neither HHV-6 DNA nor HHV-7 DNA was detected by real time PCR or nested PCR. HHV-6 DNA was detected in a single serum sample from a patient with febrile convulsions. CONCLUSION: In our study there was no association with HHV-6 or HHV-7 in most patients with central nervous system complications of influenza.     

Efficacy of human immunoglobulin and penicillin G in treatment of experimental group B streptococcal infection

In an effort to develop more effective therapy for neonatal group B streptococcal infections, penicillin G and human immune serum globulin (ISG), alone and in combination, were evaluated for their therapeutic efficacy against experimental group B streptococcal bacteremia and meningitis in newborn rats. Infected rats received either penicillin G (200 mg/kg/day), ISG (2 g/kg) or penicillin G (200 mg/kg/day) + varying doses of ISG (0.25 to 2 g/kg). All animals receiving ISG alone died; otherwise, mortality rates did not differ significantly (17-30%). Therapy with penicillin G alone and in combination with ISG was equally effective in completely eradicating group B streptococcal from blood and cerebrospinal fluid. However, combinations of penicillin G and ISG were significantly more beneficial than penicillin G alone, as shown by a significantly lower incidence of bacteremia at the end of 1 day of therapy and by greater opsonophagocytic activity in sera of animals receiving penicillin G + ISG. These findings indicate that administration of ISG in conjunction with penicillin G rapidly clears bacteria from blood, suggesting that ISG may be a useful adjunct to antimicrobial therapy of neonatal group B streptococcal disease.     

The natural history of herpes simplex virus infection of mother and newborn

The controlled evaluation of vidarabine as therapy of neonatal herpes implex virus (HSV) infection provided an opportunity to collect data to further assess the natural history of maternal and newborn infections. Women delivering infected babies were young, nulliprous, and infrequent aborters. Nearly 50% of the gestations ended in premature labor. Maternal infection was asymptomatic in 39 of 56 (70%) of the mothers, at the time of delivery. However, risk factors included a past history of genital herpes at any time and exposure to a sexual partner with presumed HSV lesions. Associated diseases in children born to these women were common. Premature infants had an incidence of respiratory distress of 52% (14 of 27). Eight of 29 (28%) term newborns had a bacterial infection, antedating the onset of neonatal HSV infection. Virologic studies on infected newborns confirmed that skin lesions were the most frequent site for virus retrieval. Progression of disease from isolated skin lesions was common, occurring in 70% of babies whose presenting sign was skin vesicles. CSF was virus-positive from 14 babies and more frequently in those with localized CNS disease. Importantly, brain biopsy was necessary for diagnosis in four cases. Finally, neither the presence or absence of antibodies to HSV was useful in predicting either presentation or outcome of infection. These studies further emphasize the complex nature of HSV infections of the newborn and need for tertiary care.     

Association of clinical presentation, laboratory findings, and virus serotypes with the presence of meningitis in hospitalized infants with enterovirus infection

One hundred eight hospitalized infants with enteroviral infections were studied to determine the association of clinical presentation, laboratory findings, and virus serotypes with the presence of meningitis. Of 108 infants, 55 (51%) had meningitis. Clinical manifestations on admission did not distinguish between infants with and those without meningitis. Echoviruses 30 and 11 and coxsackie virus B were frequently associated with meningitis (34/38; 90%) whereas echoviruses 18, 24, and 25 were not (5/35; 4%). The virus isolation rate was directly proportional to the number of leukocytes in cerebrospinal fluid: 5 of 58 (9%) when up to 9 cells/mm3 were found, 10 of 21 (48%) when 10 to 99/mm3 cells were found, and 25 of 29 (86%) when greater than equal to 100 cells/mm3 were found. Meningitis is often unsuspected in children hospitalized with enterovirus infection. The frequency of meningitis among hospitalized infants is serotype dependent and is most frequently, but not exclusively, found with pleocytosis of the cerebrospinal fluid.     

Development of acyclovir-resistant herpes simplex virus early during the treatment of herpes neonatorum.

Genotypic analysis of herpes simplex virus (HSV) DNA extracted from clinical specimens from a case of fatal disseminated neonatal HSV demonstrated that an infant developed an acyclovir-resistant HSV containing a mutation in the HSV thymidine kinase gene during the first seven days of acyclovir therapy.     

Management of Neonatal herpes simplex virus infections

As many as 2,500 infants develop neonatal herpes each year, most of whom are born to women with no history or physical findings suggestive of genital herpes. Infection usually takes one of three forms: 1) disease localized to skin, eyes, and mucous membranes, 2) localized central nervous system infection, or 3) disseminated infection. Exposure to the virus occurs during passage through an infected birth canal, but 5% of infants acquire the infectionin utero. The mortality rate is 31% for disseminated infection and 6% for localized central nervous system disease; long-term neurologic sequelae are seen in 17% and 70% of survivors, respectively. Diagnosis is made by isolating of the virus from skin lesions or other involved sites. The polymerase chain reaction for the detection of viral DNA in cerebrospinal fluid or serum is now the diagnostic test of choice for central nervous system or disseminated neonatal herpes because it has higher sensitivity than traditional culture methods. Treatment is with high-dose intravenous acyclovir (60 mg/kg per day in three divided doses), with adjustments made for infants with renal or hepatic insufficiency. Supportive measures and neuroimaging studies are often required. Acyclovir is administered for three weeks, but infants with disease localized to the skin, eyes, and mucous membranes can be treated for two weeks if the cerebrospinal fluid polymerase chain reaction assay is negative for herpes simplex virus DNA. Prevention of infection in infants can be accomplished by cesarean delivery when women have active lesions at the onset of labor. Neonates delivered through an infected birth canal should be screened between 24 and 48 hours of age with viral cultures of eyes, nasopharynx, mouth, and rectum. If positive, they should be treated with acyclovir even if asymptomatic. Suppressive acyclovir therapy beginning at 36 weeks gestation is often prescribed for women with frequent recurrences of genital herpes.