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PubMed1993

Rapid diagnosis of enterovirus infection by magnetic bead extraction and polymerase chain reaction detection of enterovirus RNA in clinical specimens.

We describe a rapid method for extraction and detection of enterovirus RNA in clinical samples. By using magnetic bead technology, enterovirus RNA was efficiently and rapidly extracted from cerebrospinal fluid, stool, saliva, blood, pericardial fluid, urine, and cryopreserved or formalin-fixed solid tissue. Enterovirus RNA was then detected by reverse transcription followed by polymerase chain reaction amplification with primers designed to allow detection of most enterovirus serotypes. For detection of enteroviruses in specimens from patients with acute enteroviral disease, the overall sensitivity of enzymatic RNA amplification was greater than that of cell culture isolation, especially in blood specimens and in stool specimens from patients with acute cardiac disease. Enterovirus RNA was also detected in cryopreserved and archival formalin-fixed myocardial tissue from patients with acute myocarditis and chronic dilated cardiomyopathy. The ability to study archival specimens is of particular value in conducting retrospective investigation. The RNA extraction procedure used was considerably faster than extraction methods using organic reagents, used less hazardous reagents, and was of similar sensitivity. This detection protocol may therefore be useful both for the diagnosis of enterovirus infection and in studying the pathogenesis of acute and chronic enterovirus-induced disease.

Adult↗
PubMed1989

Chronic relapsing pericarditis and dilated cardiomyopathy: serological evidence of persistent enterovirus infection.

By means of two different IgM-capture assays, enterovirus-specific IgM responses were shown in 9 of 14 (64%) patients with chronic relapsing pericarditis. This finding suggests persistent enterovirus infection, particularly coxsackie B virus infection. IgM responses persisted for at least 1 year and for up to 10 years after onset of symptoms. In contrast, patients with acute enterovirus infections, including acute pericarditis, had transient responses. Among patients with acute pericarditis, the level of IgM antibody was significantly higher in those who subsequently relapsed (mean 1.21, range 0.6-2.0 optical density [OD] units) than in those who did not (0.4, 0.2-0.9 OD units; p less than 0.01). Of 86 patients with dilated cardiomyopathy, 28 (33%) showed enterovirus-specific IgM responses which were present for up to 19 months before transplantation and persisted up to 4 years afterwards. Although the distribution of HLA types in these patients was similar to that in the general population, the frequency of the HLA A2 haplotype was significantly higher in those who were IgM positive. IgM antibody was significantly more common in those who had had symptoms for longer than a year before transplantation than in those with a shorter duration of symptoms (1 of 21 vs 8 of 23; p less than 0.02). Persistent virus-specific serum IgA responses were also shown in patients with chronic cardiac disease.

Acute Disease↗
PubMed1991

Influence of polyions on the early steps of enterovirus infection.

The influence of electric charged molecules on the early phases of enterovirus infection was studied in order to select antiviral compounds able to prevent viral attachment. The effect of different polyelectrolytes on the multiplication of coxsackie virus B3, echovirus 6 and hepatitis A virus was investigated in susceptible cells by adding the drug before, during or after the viral adsorption period. Among polyanions, the polysaccharides heparin and dextran sulfate inhibited viral infectivity, dextran sulfate being the most effective mainly towards hepatitis A virus infection. DEAE-dextran and protamine sulfate, generally recognized as enhancers of infectivity of naked and enveloped viruses, exhibited an inhibitory effect towards the three picornaviruses tested. Only in the case of hepatitis A did DEAE-dextran slightly improve viral antigen synthesis. The inhibitory effect shown by compounds belonging to positive and negative polyions suggests that the electric charge is not sufficient by itself to explain the antiviral activity of these drugs.

Animals↗
PubMed2004

[A case report of recurrent limbic encephalitis caused by an enterovirus infection].

We reported a 28-year-old woman with recurrent focal encephalitis caused by an enterovirus infection. She showed convulsions, abnormal behavior, and consciousness disturbance on the first admission to our hospital in July 1998. A CSF study revealed lymphocyte-dominant pleocytosis (17/microl), and brain MRI revealed high signal intensity in the bilateral limbic system including the hippocampus. We treated her with acyclovir and steroid pulse therapy based on a diagnosis of limbic encephalitis. After that, her symptoms gradually disappeared without any sequelae. She became aware of memory disturbance at the end of April 2001. She was admitted to our hospital because of generalized convulsions on 2 May 2001. Her body temperature was 37.3 degrees C. Her consciousness was disturbed (Japan coma scale 2), but there was no nuchal rigidity. A CSF study failed to reveal pleocytosis (cell count, 2/microl). Brain MRI revealed a bilateral high signal intensity area in the limbic system. Her symptoms gradually improved, but the mild memory disturbance persisted on her discharge. An immunological study revealed no abnormality in either humoral or cell immunity. RT-PCR revealed enterovirus RNA in CSF samples obtained on 1 July 1998, 3 May 2001, and 25 July 2001. We diagnosed her as having recurrent limbic encephalitis caused by an enterovirus infection. This is the first report of recurrent limbic encepalitis caused by enterovirus infection in an host with an undetected defect of immune system.

Adult↗
PubMed2004

Intrauterine exposure to maternal enterovirus infection as a risk factor for development of autoimmune thyroiditis during childhood and adolescence.

Maternal intrauterine enterovirus infection during pregnancy increases the risk for the offspring to develop type 1 diabetes mellitus. Type 1 diabetes mellitus and autoimmune thyroiditits (AIT) are closely linked. A common pathogenetic factor is possible. The objective of this study was to investigate a possible association between maternal enterovirus infection during pregnancy and the development of AIT in the offspring. Sera taken at delivery from 31 mothers whose children subsequently developed AIT was analyzed for immunoglobulin (Ig)A, IgG, and IgM antibodies against enterovirus, and compared to a control group comprising 233 randomly selected maternal sera. Of the mothers whose children developed AIT, 5 of 31 (16%) were enterovirus IgM-positive, compared to 17 of 233 (7%) in the control group (p = 0.16). The age at diagnosis of AIT was significantly lower in the group of children with IgM-positive mothers compared to children with IgM-negative mothers (p < 0.05). In addition, 3 children (60%) in the IgM-positive group were overtly hypothyroid at diagnosis of AIT, compared to no child (0%) in the IgM-negative group (p < 0.01). No significant differences were found in IgA and IgG antibody titers between the mothers whose children developed AIT and the control group. Although this study did not have enough power to reveal intrauterine exposure to maternal enterovirus infection during pregnancy as a risk factor for development of AIT during childhood and adolescence, it suggested an association with earlier onset of clinical disease in children to enterovirus IgM-seropositive mothers.

Adolescent↗
PubMed2003

Establishing evidence for enterovirus infection in chronic disease.

Viruses have long been considered among potential environmental triggers of type 1 diabetes mellitus. Epidemiologic and seroprevalence studies have associated enterovirus infection with development of prediabetic autoimmunity and with the onset of clinical diabetes. Enterovirus infection has also been temporally correlated with disease onset by virus isolation or by detection of viral genome by reverse transcription-polymerase chain reaction (RT-PCR). For the large-scale prospective studies that are required to firmly establish a causal relationship between enterovirus infection and development of prediabetic autoimmunity or progression from autoimmunity to clinical diabetes, sensitive RT-PCR methods must be used to detect virus prior to the onset of diabetic symptoms. We have developed an RT-seminested PCR protocol to detect enteroviruses in clinical specimens. This method is approximately 10,000-fold more sensitive than conventional, single-amplification PCR. Further, we have developed molecular methods to rapidly and reliably identify enterovirus serotype, bypassing the cumbersome and often problematic neutralization test. The molecular serotyping approach will be valuable in examining the relationships between particular virus serotypes or genotypes and specific diseases.

Chronic Disease↗
PubMed1991

[Coxsackie B enterovirus infection in Astrakhan Province].

Overall 132 patients with Coxsackie B enteroviral infection were admitted to an Astrakhan infectious hospital in 1989. Of these, 100 patients had aseptic meningitis and 32 presented with respiratory infection syndrome. The severity of enteroviral meningitis may vary from mild forms running its course without marked meningeal syndrome to grave ones associated with high pleocytosis, manifest syndrome of CSF hypertension and potential exacerbation of the infectious process. The patients with the syndrome of respiratory infection demonstrated its combination with intestinal disorders (39%). The ECG changes attested to marked toxic alterations in the myocardium. The peripheral blood showed leukopenia with relative lymphocytosis. The treatment and future course of the disease are dependent on a correct etiological diagnosis which is feasible with joint efforts of clinicians, epidemiologists and virologists.

Acute Disease↗
PubMed1992

[The relationship of clinical manifestations and age distribution in enterovirus infections--viral isolation and seroepidemiology in Aichi Prefecture].

We studied the epidemiology of enterovirus infection in Aichi Prefecture from 1985 to 1989. We examined the age distribution of aseptic meningitis patients (AM) and exanthematous disease patients (Ex) and a seroepidemiological study of echovirus type 7 (E7), E9, E18 and group A coxsackievirus type 9 (CA9), was performed. The results is as follows: 1) E7, E9, E18 and CA9 were isolated from AM and Ex but E6, E11 and group B Coxsackie viruses (CB) were isolated in fewer cases from Ex. 2) The AM was consistently increased from June to August. Whereas the Ex was seen in all seasons but a slight increase was noted between June to July, and enteroviruses (EV) isolation was increased in this season. 3) The AM occurred in 0 year olds and 4 year olds whereas the Ex was seen in 0 to 1 years. EV was mainly isolated from 0-1 year olds. 4) The relationship of clinical manifestations and age was very clear in E9 and E18, a higher proportion of children at 1 years or under were those of the Ex and most children of the latter part of 4 years were those of the AM. The Ex had the same results with E7 and CA9 but AM was increased in 0 years and 4 year olds. 5) We studied the age distributions of neutralizing antibodies against E7, E9, E18 and CA9. The positive rate of neutralizing antibody after prevalence rose between 2-5 years of age. There were few patients among the 2 to 3 year olds but the neutralizing antibody was raised in this age. I considered that reason the enteroviruses infected mainly the 2 to 6 year olds showing no clinical symptoms where as some of the 3 year olds had aseptic meningitis and some under 1 year had symptoms of exanthematous diseases.

Age Factors↗
PubMed2002

Enterovirus infections and type 1 diabetes.

Type 1 (insulin-dependent) diabetes is a typical organ-specific autoimmune disease where insulin-producing beta cells are destroyed by immune mediated mechanisms. The risk of the disease is modulated by genetic factors, mainly genes coding for human leukocyte antigens (HLA), but environmental factors are needed to trigger the process in genetically susceptible individuals. Possible viral triggers of the disease have been sought for years but their identification has been very difficult. Recently, considerable progress has been made by employing new research methods which have supported the idea that the group of enteroviruses may be particularly important in the pathogenesis. An association between enterovirus infections and type 1 diabetes was first reported 30 years ago and since then evaluated in several studies. Recent molecular studies have considerably strengthened this hypothesis by showing that enterovirus genome is present in the blood of diabetic patients. In addition, the first prospective studies have suggested that enterovirus infections may initiate the beta-cell damaging process several years before clinical diabetes is diagnosed. Ecological studies have also indicated similarities in the epidemiology of type 1 diabetes and poliomyelitis - a well-known enterovirus disease. Experimental models, like enterovirus-infected mice or in vitro-cultured beta cells, have provided important information about possible mechanisms, but still it is not known how beta cells are destroyed in human beings. The ongoing prospective studies will answer many open questions, and should the association still hold true, intervention trials will be needed to confirm causality. Even if enterovirus infections were not associated with all diabetes cases but rather with a subgroup of them, this would offer attractive possibilities to prevent the disease or part of it, for example, by an enterovirus vaccine.

Animals↗
PubMed1993

Increased transcription of interleukin-6 in the brains of mice with chronic enterovirus infection.

An animal model of chronic enteroviral infection was established by using PCR to detect viral genomes in animal tissues and to compare levels of transcription of a variety of cytokines in the brain. Chronic coxsackie-virus B1 infection was found in both brain and skeletal muscle of mice infected as neonates. The viral infection cleared by 240 days post-infection. Elevated levels of tumour necrosis factor alpha and interleukin-6 (IL-6) would appear to be linked to acute and chronic infection respectively. Levels of IL-6 return to normal upon clearance of the virus.

Animals↗
PubMed1975

Oral polio vaccination of children in the tropics. III. Intercurrent enterovirus infections, vaccine virus take and antibody response.

The effect of intercurrent enterovirus infections on host responses to oral polio vaccine (OPV) was studied in groups of infants and children who were without antibodies to one, two or three serotypes of poliovirus. The prevalence of enterovirus infections as detected in fecal specimens collected at weekly intervals and inoculated in primary monkey kidney cell culture, HEp 2 cells and newborn mice ranged between 60 and 70 per cent. The presence of such infections at the time of, 1 week prior to, or during the 3 weeks prior to the administration of OPV did not appear to inhibit either vaccine virus take or antibody response. However, in both the infected and uninfected children the rates of vaccine virus take and seroconversion were found to be considerably lower than those reported from several temperate climate countries.

Child, Preschool↗
PubMed1997

Islet cell antibody seroconversion in children is temporally associated with enterovirus infections. Childhood Diabetes in Finland (DiMe) Study Group.

Exposure to Coxsackie B virus or other enteroviruses prenatally or in childhood increases the risk for later manifestation of insulin-dependent diabetes mellitus (IDDM). The occurrence of enterovirus infections was analyzed in 23 initially nondiabetic and islet cell antibody (ICA)-negative siblings of IDDM patients who converted to ICA positivity during a prospective follow-up study. Increases in enterovirus antibody levels, documented by heavy chain-capture RIA and EIA techniques, were significantly more frequent in sample intervals in which ICA first appeared (18/23, 78%) than in other sample intervals in these siblings (30/92, 33%; P < .001) or all sample intervals in 97 ICA-negative control siblings (117/403, 29%; P < .001). The children who converted to ICA positivity during an enterovirus infection more often had the high-risk HLA-DQB1 genotype than did children who were constantly ICA-negative (P < .01). The results suggest that enteroviruses may be important in the induction of a beta cell damaging process long before the clinical manifestation of IDDM.

Adolescent↗
PubMed1993

Enterovirus infections in neonates. A retrospective study of 21 cases.

OBJECTIVES: The aim of this study was to define the major features of enterovirus infections in the neonatal period based on our own experience. METHODS: Epidemiology, clinical manifestations and laboratory investigations concerning 21 neonates having experienced a Coxsackie B or an Echovirus infection between 1987 and 1991, were retrospectively reviewed. Aetiological diagnosis was made by classical viral isolation and/or by evidencing Coxsackie B-specific IgM antibodies with an immunocapture enzyme immunoassay. RESULTS: In 13 neonates the infection occurred between June and September. The onset of clinical signs ranged from day 1 to day 25 after birth with two separate periods: before 7 days of age, suggesting a perinatal transmission of the virus, or beyond this date, more likely connected with a postnatal transmission. Clinical manifestations included hyperthermia, gastroenteritis, meningitis, encephalitis, pneumonia and myocarditis, with a diphasic pattern in 6 cases. Most of the neonates improved gradually and developed normally. The Coxsackie B-specific IgM assay was the most rapid method whereas viral isolation, even though it took more time, was the most sensitive technique to establish the aetiological diagnosis in neonates. CONCLUSIONS: Enterovirus infections in neonates are difficult to diagnose and to differentiate from bacterial infections. A viral-like illness in the environment of the neonate allows the clinician to anticipate the clinical signs and a possibly fatal disease. Identification of the causal virus should be performed by both viral isolation and search for specific IgM antibodies. Treatment and prophylaxis are so far disappointing.

Antibodies, Viral↗