Echovirus Type Research Articles

Nucleic acid sequence relationships between enterovirus serotypes

Forty-eight different enterovirus serotypes were analysed by a nucleic-acid hybridization test using probes derived from the 3′ end of coxsackievirus A21 (CA21) and B3 (CB3), poliovirus 3 (P3) and enterovirus 70 (E70). More than 90% of the serotypes could be detected with this collection of reagents. The CB3 probe reacted with all the coxsackie B viruses, with all three poliovirus serotypes, and with almost all of the 30 ECHO virus types tested. In addition some of the coxsackie A viruses and the BrDr 73 strain of enterovirus 71 gave a positive signal with this probe. The P3 probe detected all the poliovirus strains and also some coxsackievirus A isolates but no coxsackie B or ECHO viruses. A similar hybridization pattern as with the P3 probe was obtained when the CA 21 probe was used. The E70 probe appeared to be strain-specific. The results indicate that nucleic-acid hybridization is a useful method for rapid detection and subgrouping of enteroviruses during virus isolation, and that the test could be further developed for typing of the strains.

Viral antibodies in agricultural populations exposed to aerosols from wastewater irrigation during a viral disease outbreak.

The presence of antibodies to eight enteroviruses (echovirus types 4, 7, and 9, coxsackievirus types A9, B1, B3, and B4, and hepatitis A virus) and varicella-zoster virus was determined during a two-year period, 1980-1981, in paired blood samples of 777 persons in selected agricultural communities (kibbutzim) in Israel. These communities were divided into several categories on the basis of wastewater utilization for sprinkler irrigation and/or fish ponds. Among the nine viral antibodies studied, there was a consistent and significant excess of antibodies to echovirus type 4 only, particularly in the age group 0-5 years, in kibbutzim that had been exposed to aerosols from sprinkler irrigation with partially treated wastewater from nearby towns. This finding may be attributed to a major national echovirus type 4 epidemic, which had peaked shortly before the collection of the blood samples. The fact that no similar excess of the other viral antibodies studied was found in any of the kibbutz categories suggests that, under nonepidemic conditions, exposure to wastewater aerosols usually does not lead to an excess in enteroviral infection. No excess of clinical cases of echovirus type 4-associated disease (meningitis or encephalitis) was detected in the communities exposed to wastewater aerosol.

Direct inactivation of viruses by human granulocyte defensins.

Human neutrophils contain a family of microbicidal peptides known as defensins. One of these defensins, human neutrophil peptide (HNP)-1, was purified, and its ability to directly inactivate several viruses was extensively tested. Herpes simplex virus (HSV) types 1 and 2, cytomegalovirus, vesicular stomatitis virus, and influenza virus A/WSN were inactivated by incubation with HNP-1. Two nonenveloped viruses, echovirus type 11 and reovirus type 3, were resistant to inactivation. Purified homologous peptides HNP-2 and HNP-3 were found to have HSV-1-neutralizing activities approximately equal to that of HNP-1. Inactivation of HSV-1 by HNP-1 depended on the time, temperature, and pH of incubation. Antiviral activity was abrogated by low temperature or prior reduction and alkylation of the defensins. Addition of serum or serum albumin to the incubation mixtures inhibited neutralization of HSV-1 by HNP-1. We used density gradient sedimentation techniques to demonstrate that HNP-1 bound to HSV-1 in a temperature-dependent manner. We speculate that binding of defensin peptides to certain viruses may impair their ability to infect cells.

Survival and Transport of Hepatitis a Virus in Soils, Groundwater and Wastewater

Hepatitis A virus (HAV), poliovirus type 1 and echovirus type 1 were studied for their adsorption and survival in groundwater, wastewater and soils suspended in these media and for survival and transport through unsaturated miniature soil columns intermittently dosed with virus-laden groundwater or wastewater. There were differences among the viruses in adsorption to soils, with poliovirus adsorbed most extensively, echovirus the least and HAV intermediate between these two. All three viruses survived well (>90% inactivation) for at least 12 weeks in groundwater, wastewater and soil suspensions at 5°C. However, at 25 C, HAV survived generally longer than poliovirus and echovirus, with 90–99% inactivation of HAV and 99.9–99.99% inactivation of poliovirus and echovirus in 12 weeks. In miniature soil columns dosed with virus-laden groundwater or wastewater, virus reductions were generally least for echovirus and greatest for poliovirus. HAV reductions were intermediate between these two, but more like poliovirus. The ability of HAV to survive for long periods in soils, groundwater and wastewater and to migrate to some extent through unsaturated soils helps to explain why HAV can contaminate groundwater and cause outbreaks of groundwaterborne disease.

Use of WIN 51711 to prevent echovirus type 9-induced paralysis in suckling mice.

The systemic efficacy of the antipicornavirus agent WIN 51711 was assessed in suckling mice infected with echovirus type 9 (Barty strain). Single, daily intraperitoneal doses of WIN 51711 as low as 10 mg/kg significantly (P less than .01) slowed the rate of onset of echovirus-induced paralysis and reduced the number of paralyzed animals. Intraperitoneal administration beginning 2 hr before infection or 48 hr after infection was equally effective in preventing paralysis. Oral administration of WIN 51711 twice daily beginning 72 hr after infection was the most-effective dosage regimen, with doses as low as 3 mg/kg preventing paralysis in 75% of the animals. Titers of virus in asymptomatic mice on day 6 after infection were reduced by up to 4.75 log pfu in WIN 51711-medicated mice when compared with placebo. Maximal concentrations of WIN 51711 in adult mice after oral medication with a 100 mg/kg dose were 24.3, 21.5, 10.4, 9.8, 6.9, and 4.1 micrograms/g in heart, kidney, brain, liver, serum (micrograms/ml), and muscle, respectively at 0.5-1.0 hr after medication.

Rapid diagnosis of echovirus 33 infection by neutralizing specific IgM antibody.

In 1982, we isolated 20 strains of echovirus type 33 (EV33) from 18 patients. We studied the humoral response to EV33 of 2,437 subjects from whom at least one serum was available during the same year. In 388 subjects with the neutralizing antibody level at 64 or more, we assayed the EV33 IgM antibodies by seroneutralization test after fractionation of sera by ion exchange chromatography. One hundred ninety-five subjects (8.0%) had a high titre (greater than or equal to 32) of EV33 IgM antibodies, which was considered as evidence of recent infection. The EV33-positive IgM fractions were assayed against five other enteroviruses. Sixty percent of the IgM fractions did not cross-react with any of the five serotypes, 8.7% cross-reacted with at least one serotype but with predominant EV33 IgM response, and 31.3% had an equivalent or greater amount of non-EV33 IgM antibodies; the type specificity of the assay was directly related to the age of the subjects. These findings suggest that determination of neutralizing specific IgM antibody is a sensitive and rapid test for the diagnosis of enterovirus infections, especially in young people.

Characterization of an Echovirus Type 30 Variant Isolated from Patients with Aseptic Meningitis

An outbreak of echovirus type 30 infection associated with aseptic meningitis occurred among newborn babies in a hospital neonatal room at Fukui City in 1983. The isolated virus was identified as an antigenic variant of echovirus type 30 by cross-neutralization tests with antisera against the prototype Bastianni strain and the present isolate. Western blot analysis demonstrated that the antigenic determinants responsible for virus neutralization, some of which were type specific and others strain specific, were present on the capsid protein VP1. The current strain was more thermoresistant than the prototype, suggesting some alterations in virus structural proteins.

An epidemic of acute diarrhoea in rural southern India associated with echovirus type 11 infection.

An epidemic of diarrhoea with two distinct waves affected a village of 1375 people in southern India in 1983. The first wave of the epidemic, from the last week of December 1982, had a sharp peak in January 1983 and was over by March. Echovirus type 11 was isolated from patients, who also had a serum antibody response to the virus. During the second wave of the epidemic, from May to September 1983, the clinical features were different and Shigella flexneri was isolated without significant viral isolates. Infection during the first wave did not protect from the second wave. Virus isolation was in human intestinal tumour-derived differentiated epithelial cell lines; such cell lines may be useful for the isolation and identification of enteroviruses in clinical samples.

Concentration of Seeded and Naturally Occurring Enteroviruses from Waters of Varying Quality by Hollow Fiber Ultrafiltration

A semi-automated hollow fiber ultrafiltration system was used to concentrate viruses from tap water, river water and treated and untreated wastewater. Samples were dehydrated by filtration through an asymmetric polysulfone hollow fiber module with a molecular weight cut-off of 10,000 D. Retained solute was recovered from the filter by backwashing with water, buffer and other solutions. One hundred liters of tap water seeded with poliovirus type 2 Sabin were processed in 161 ± 22 minutes (average flux 1.2 ± 0.1 × 10−3 cm/sec). Retained virus was recovered from the filter by backwashing with 150 ml of 0.05 M glycine, pH 10.0, which was further reduced to about 10 ml by the organic flocculation technique. Average recovery of 60 plaque-forming units (PFU) of poliovirus from 100 1 of tapwater was 148 ± 10%. Hudson River water (100 1 with an average turbidity of 3.2 NTU) was processed in 218 ± 34 minutes (average flux 0.8 ± 0.2 × 10−3 cm/sec). Recovery of 114 PFU of poliovirus seeded into 99 1 of river water was 35%. No natural virus was recovered from unseeded river water. Naturally occurring viruses, tentatively identified as echovirus type 14, coxsackie virus type A9 and coxsackie virus types B2-B5, were isolated from concentrates qf treated and untreated wastewater (average fluxes 0.58 × 10−3 and 0.46 × 10−3 cm/sec, respectively).

NMR parameters of local anesthetics as biological markers of the cell-virus interactions.

The selective proton NMR relaxation rates were measured in the system made by the local anesthetic procaine, human erythrocytes and the hemagglutinating Echo virus type 11. The NMR technique provided a very good tool for using drugs firmly bound to receptor membranes as biological "markers" of the virus-cell interaction.

Prevention of spread of echovirus 6 in a special care baby unit.

A case of fatal neonatal infection with enteric cytopathogenic human orphan virus (echovirus) type 6 is presented. The measures taken to prevent further spread of infection with special reference to the use of human normal immunoglobulin are described.

Direct inactivation of viruses by MCP-1 and MCP-2, natural peptide antibiotics from rabbit leukocytes.

Six homologous peptides were purified to homogeneity from rabbit granulocytes or alveolar macrophages and tested for their ability to inactivate herpes simplex virus type 1 (HSV-1). Two of the peptides, MCP-1 and MCP-2, showed considerable in vitro neutralizing activity, whereas four structurally homologous peptides (NP-3a, NP-3b, NP-4, and NP-5) were relatively ineffective. Inactivation of HSV-1 by MCP-1 or MCP-2 depended on peptide concentration and on the time, temperature, and pH of the incubation. HSV-2, vesicular stomatitis virus, and influenza virus A/WSN were also susceptible to direct neutralization by MCP-1 or MCP-2, whereas cytomegalovirus, echovirus type 11, and reovirus type 3 were not. We speculate that MCP-1 and MCP-2, peptides that are abundant in rabbit granulocytes and lung macrophages, may contribute to antiviral defenses by mediating the direct inactivation of HSV-1 and selected other viruses.

Virion conformational forms and the complex inactivation kinetics of echovirus by chlorine in water.

Aberrant inactivation kinetics were observed when monodispersed echovirus type 1 (Farouk) was inactivated with chlorine. An initial 1- to 2-log10-unit decrease in titer was followed by lag period, during which the titer stayed the same or increased, and this was followed by a final decline in titer. First-order kinetics were obtained with poliovirus type 1 under the same conditions. Isoelectric focusing studies of echovirus before chlorine treatment showed that the virus distributed into two pH-dependent and interconvertible isoelectric forms. After chlorine treatment all remaining virus infectivity was associated with a third pH-independent isoelectric form. The complex inactivation kinetics appeared to be due to shifts between these conformational forms during inactivation in certain ionic environments. Under certain conditions the conformational shifts resulted in substantial resistance of monodispersed echovirus to chlorine.

Seasonal distribution of enteroviruses and adenoviruses in domestic sewage.

A seasonal distribution of enteroviruses and adenoviruses in raw sewage effluents of Athens, Greece, was observed over a 15-month surveillance period. All 36 samples tested were positive for both virus groups. Adenovirus concentration levels ranged from 70 to 3200 cytopathic units per litre of sample, whereas the corresponding values for enteroviruses were 90-900 cytopathic units per litre. Peak values for adenoviruses were recorded during the months of April and June 1983, whereas for enteroviruses the peak was recorded in September 1983. All three types of poliovirus were present. Coxsackievirus types B-1, B-2, B-4, and B-5 and echovirus type 7 were also isolated. Adenovirus types 1, 2, 3, 5, 7, and 15 were detected as well.

Rapid diagnosis of echovirus type 33 meningitis by specific IgM detection using an enzyme linked immunosorbent assay (ELISA)

During an outbreak of meningitis in France (in the Lyon area), from June to October 1982, serum and stool samples were collected from 227 patients. An enzyme-linked immunosorbent assay (ELISA) for titrating IgG and IgM antibodies anti-echovirus type 33 was developed and compared with the virus isolation technique, and with the titration of neutralizing antibodies. In 39 patients excreting echovirus 33 in faeces, the ELISA test allowed a positive serodiagnosis in 85% of the cases by detection of specific IgM (64% of the cases) and by seroconversion (21%). Compared with the neutralization (Nt) test, ELISA was found to be more sensitive. The antibody titres in ELISA were over 50 times higher and detected earlier than the neutralizing antibodies. This early immune response allowed a rapid diagnosis by specific IgM detection in the acute sera collected within 8 days after the appearance of the clinical symptoms in more than 50% of the 97 patients examined, whereas the Nt test allowed a positive serodiagnosis in only 32% of the patients. The use of a caesium chloride purified antigen insured the specificity of the reactions.

Enteroviruses and acute encephalopathy syndrome in Nagpur.

The role of enteroviruses in the etiology of an acute encephalopathy syndrome of infants and children, prevalent in Nagpur since 1958, was investigated in 1967 and 1968. Specimens of various body fluids or tissues were collected from 255 patients and examined for the presence of enteroviruses by inoculation in cell culture and in newborn mice. One or more specimens from 20 patients (8%) were found positive. The virus isolates included echovirus type 7 (7 patients), coxsackie B2 (3 patients), poliovirus types 1, 2 and 3, echovirus types 1 and 9, coxsackie B1 and several untypable strains. Some patients had dual virus infections. Supported by other available information on the role of enteroviruses in the etiology of Reye’s and similar syndromes, it is postulated that enteroviruses are the causative agents only in a small proportion of the cases of the Nagpur encephalopathy syndrome.

Immunoglobulin responses to echovirus type 11 by enzyme linked immunosorbent assay: Single-serum diagnosis of acute infection by specific IgM antibody

An indirect solid-phase enzyme-linked immunosorbent assay (ELISA) for the determination of specific IgM and IgG antibodies to echovirus type 11 in a single dilution of serum was developed using partially purified echovirus type 11 bound to microplates. Whole serum was used for IgG antibody but prior to assaying for IgM antibody interfering IgG was removed by ion exchange chromatography. The ELISA for echovirus type 11 IgG antibody was a more sensitive, rapid, technically easier and less costly alternative to the neutralisation test. With the IgG ELISA 12 of 132 sera (10.6%) known to contain enterovirus antibodies other than echovirus type 11 were positive but it could not be determined to what extent this was due to the greater sensitivity of the ELISA or cross-reactions. The IgM ELISA was even more sensitive than the IgG ELISA with acute sera, and showed a reactivity in 4 of 36 sera (11.1%) with no detectable echovirus type 11 neutralising antibodies. Echovirus type 11 IgM antibody was detected in all sera collected after the first week of infection and up to 30 days after infection. However, it was only detected in 58% of sera collected during the first week after onset thus limiting its use for rapid diagnosis. The echovirus type 11 IgM ELISA appears to have considerable laboratory diagnostic potential when a rising antibody level cannot be demonstrated in paired sera or when virus is not cultured.

Fatal hepatoadrenal necrosis in the neonate associated with echovirus types 11 and 12 presenting as a surgical emergency

Two neonates presented to our service with suspected intraabdominal surgical emergencies. Both subsequently were found to have hepatoadrenal necrosis due to overwhelming echoviral infection. Echovirus types 11 and 12 were isolated postmortem. The latter virus has not been associated previously with fulminant neonatal illness.

Echovirus type 11 infection in neonates in Fiji

Echovirus type 11 infection in neonates in Fiji

Acquisition of antibodies to various Coxsackie and Echo viruses and hepatitis A virus in agricultural communal settlements in Israel.

A seroepidemiological study was conducted to measure the antibody prevalence for eight different enteric viruses. These include seven "classical" enteroviruses, ie, Coxsackie virus types A9, B1, B3, B4 and three ECHO virus types 4, 7, and 9, as well as hepatitis A virus (HAV), recently classified as enterovirus 72. Sera samples were obtained from 791 residents living in 29 agricultural settlements (kibbutzim) with a total population of 15,950. The results indicated that the acquisition of antibodies to Coxsackie and ECHO viruses occurred at a significantly earlier age than to HAV. Prevalence of antibodies to Coxsackie and ECHO viruses at two to four years of age was 40%-69%, in contrast to 4% for HAV. At age 5-17 years, 85% had antibodies to five or more of the seven "classical" enteroviruses, as compared with 10% to HAV. The latter steeply increased to 63% for age group 18-24 years and reached a peak of 95% for age group 50 years and older. Prevalence of antibodies to ECHO and Coxsackie viruses were often found to be higher in in females than in males, within the age group of 18 years and older, but not in age groups 18 years and younger. Prevalence of antibodies to HAV was observed to be much higher in males than in females 18-24 years of age. No significant difference was found in the prevalence of individuals harboring antibodies to three or more and even up to five or more Coxsackie and ECHO viruses between those HAV-negative versus those HAV positive.(ABSTRACT TRUNCATED AT 250 WORDS)