Identification of vaccine-related polioviruses by hybridization with specific RNA probes.

We developed RNA probes for the identification of poliovirus isolates by blot hybridization. Two sets of vaccine strain-specific probes were prepared. They complemented variable genomic domains within (i) the 5'-untranslated region and (ii) the amino-terminal codons of VP1.

Paralytic poliomyelitis in Romania, 1984-1992. Evidence for a high risk of vaccine-associated disease and reintroduction of wild-virus infection.

Although poliomyelitis due to wild-virus infection has virtually disappeared from Romania, with no cases having been documented between 1984 and 1989, vaccine-associated paralytic poliomyelitis has been reported at very high rates for over two decades. In November 1990, to decrease the risk of vaccine-associated paralytic poliomyelitis, oral poliovirus vaccine produced in Romania was replaced by imported oral vaccine made by a Western European manufacturer. To better quantify the risk of vaccine-associated paralytic poliomyelitis and the impact of the change in vaccine manufacturer, the authors reviewed clinical, epidemiologic, and laboratory data on poliomyelitis cases that occurred in Romania from 1984 to 1992.

Vaccine-preventable disease susceptibility in a young adult Micronesian population.

Current US military recruit vaccination policy presumes that recruits have had a complete childhood immunization series. This assumption may not be appropriate for recruits from Micronesia, who may have had limited access to modern health care, including immunization programs. During 1988 and 1990, a cross-sectional serosurvey was conducted among 66 US military recruits, 56 from the Federated States of Micronesia and 10 from the Republic of the Marshall Islands, collectively referred to as Micronesia.

Echovirus 30 infection and aseptic meningitis in parents of children attending a child care center.

In July 1992, 13 parents with children attending a child care center (CCC) developed aseptic meningitis (AM) due to echovirus 30 (E30). To determine the extent of illness and risk factors for transmission, survey and blood specimens were collected from CCC families and teachers and from adult and pediatric controls. Infection was defined as the presence of anti-E30 IgM antibodies.

Enterovirus 71 infections and neurologic disease--United States, 1977-1991.

Since first described in 1974, enterovirus 71 infections have been associated with severe neurologic disease, and widespread infection was suspected in 1987. To investigate enterovirus 71 activity further, data were reviewed for isolations reported nationally during 1977-1991, virology laboratories were contacted regarding isolations during 1985-1989, and medical records were reviewed for respective patients, 1985-1989. From 1977 to 1991, 193 culture-confirmed enterovirus 71 infections were identified: > or = 1 isolate each year.

Direct detection of wild poliovirus circulation by stool surveys of healthy children and analysis of community wastewater.

Cartagena, Colombia, was one of the last cities in the Americas known to have endemic poliomyelitis. After 3 cases were identified in 1991, two approaches for detecting continued silent transmission of wild polioviruses within a high-risk community were used: stool surveys of healthy children and virologic analysis of community sewage. Wild type 1 polioviruses were isolated from 8% of the children studied and from 21% of sewage samples.

Distribution of wild type 1 poliovirus genotypes in China.

Poliomyelitis remains an important public health problem in China. Most cases and outbreaks are associated with wild type 1 polioviruses. To obtain an overview of type 1 poliovirus transmission in China, partial genomic sequences were compared for 24 case isolates from 12 provinces.

Paralytic poliomyelitis in Oman: association between regional differences in attack rate and variations in antibody responses to oral poliovirus vaccine.

Variation in attack rates of paralytic disease by region during the 1988-1989 epidemic of type 1 poliomyelitis in Oman provided the stimulus to test the hypothesis that these observations were due to regional differences in the response of infants to trivalent oral poliovirus vaccine (OPV). Seroprevalence studies of 394 children born during the outbreak were conducted in six different regions of Oman and in two socioeconomic status (SES) groups in the capital city of Muscat; a seroconversion study was also carried out in 105 infants born after the outbreak. Seroprevalence rates by region after receipt of at least three doses of OPV ranged from 90% to 100% (median 94%) to poliovirus type 1, and from 86% to 100% (median 97%) to type 2, and from 47% to 79% (median 72%) to type 3, with the lowest rates observed in regions with the highest incidence of type 1 paralytic disease.

Multiple primer pairs for polymerase chain reaction (PCR) amplification of human parvovirus B19 DNA.

Human parvovirus B19 is the etiologic agent of erythema infectiosum and transient aplastic crisis in patients with hemolytic anemias and has been associated with fetal death, arthritis, and chronic anemia. Acute B19 infection is best diagnosed by detection of IgM antibodies, whereas the diagnosis of chronic infection often requires the sensitivity of PCR to demonstrate presence of virus over time. To improve our ability to detect B19 DNA by polymerase chain reaction (PCR), we evaluated 19 primers combined into 16 different primer pairs for their ability to detect temporally and geographically diverse B19 isolates.

Immunogenicity of a supplemental dose of oral versus inactivated poliovirus vaccine.

In many developing countries, the immunogenicity of three doses of live, attenuated, oral poliovirus vaccine (OPV) is lower than that in industrialised countries. We evaluated serum neutralising antibody responses in 368 children aged 6 months and 346 children aged 9 months in Côte d'Ivoire who had previously received three doses of OPV at 2, 3, and 4 months of age, and who were then randomised to receive a supplemental dose of OPV or enhanced-potency inactivated poliovirus vaccine (IPV) at the time of measles vaccination. Although both vaccines increased seroconversion to all three poliovirus types, antibody responses were greater in the IPV group.

Coxsackievirus B2 infection and aseptic meningitis: a focal outbreak among members of a high school football team.

In an investigation of a school outbreak of enterovirus-like illness and aseptic meningitis, an IgM antibody assay was used to identify persons with evidence of recent coxsackie virus B2 infection. During September and October 1989, 81 (25%) of 319 students and staff reported an enterovirus-like illness; of these, 17 (21%) also had aseptic meningitis. Attack rates for enterovirus-like illness were highest among varsity football team members (53%), and most of this illness occurred between 6 and 15 October.

Acute hemorrhagic conjunctivitis due to enterovirus 70 in American Samoa: serum-neutralizing antibodies and sex-specific protection.

Acute hemorrhagic conjunctivitis due to enterovirus 70 has caused extensive outbreaks in tropical areas since 1969. Between December 1, 1990, and March 4, 1991, an outbreak of acute hemorrhagic conjunctivitis due to enterovirus 70 occurred in American Samoa, where an outbreak due to the same agent had occurred in 1981. A survey of 5% of the households (134 households, 1,095 individuals) was conducted throughout the island of Tutuila.

Genotype-specific in vitro amplification of sequences of the wild type 3 polioviruses from Mexico and Guatemala.

The extensive nucleotide sequence heterogeneity among independent genotypes of wild polioviruses permits the systematic design of genotype-specific molecular reagents. We have prepared two sets of polymerase chain reaction (PCR) primer pairs specific for the genotype of wild poliovirus type 3 recently endemic to Mexico and Guatemala. Nucleotide sequences of a representative wild type 3 virus isolated in Mexico in 1989 differed from the corresponding Sabin 3 (Leon 12 a1b) sequences at 167 of 900 positions within the VP1 region.

[Epidemic outbreak of viral meningitis caused by type 30 ECHO virus].

The clinical records of 15 children admitted to Hospital Infantile de México Federico Gómez with diagnosis of viral meningitis were reviewed. They were part of 19 patients admitted with this diagnosis during a 5 week period (March 22 to April 30, 1992) and represent a significant increase of aseptic meningitis compared with the same periods of previous years at Hospital Infantile de Mexico and in Mexico City where there is an ongoing epidemic outbreak of this entity. All the patients studied had spinal fluid findings consistent with viral meningitis and in 4 of them on ECHO virus type 30 was isolated at the Enterovirus Section of the CDC, Atlanta Georgia USA.

Using the virus challenge dose in the analysis of virus neutralization assays.

We propose a new basis for adjusting the results of virus neutralization assays. These assays consist of two separate experiments performed in parallel: a virus titration experiment and a serum dilution assay. In the virus titration experiment, one estimates the amount of virus used in the assay (the virus challenge dose).

Immunoglobulin class and subclass-specific monoclonal antibody sandwich ELISA for the detection of antibodies against coxsackieviruses B, types 1-5.

Immunoglobulin subclass-specific ELISAs were developed for human IgG1, IgG2, IgG3, IgG4, IgAtotal, and IgM directed against Coxsackie B (CB) virus types 1, 2, 3, 4, and 5. In all the assays the solid phase was coated with immunoglobulin class/subclass-specific monoclonal antibodies, followed by an incubation with the serum specimens. Incubation with one of the CB viruses, as well as an incubation with biotinylated serotype-specific monoclonal antibodies to the same virus type provided the virus specificity.

Epidemiology of poliomyelitis in the United States one decade after the last reported case of indigenous wild virus-associated disease.

Poliomyelitis caused by wild poliovirus has been virtually nonexistent in the United States since 1980, and vaccine-associated paralytic poliomyelitis (VAPP) has emerged as the predominant form of the disease. We reviewed national surveillance data on poliomyelitis for 1960-1989 to assess the changing risks of wild-virus, vaccine-associated, and imported paralytic disease; we also sought to characterize the epidemiology of poliomyelitis for the period 1980-1989. The risk of VAPP has remained exceedingly low but stable since the mid-1960s, with approximately 1 case occurring per 2.

Outbreak of paralytic poliomyelitis in Oman: evidence for widespread transmission among fully vaccinated children.

From January, 1988, to March, 1989, a widespread outbreak (118 cases) of poliomyelitis type 1 occurred in Oman. Incidence of paralytic disease was highest in children younger than 2 years (87/100,000) despite an immunisation programme that recently had raised coverage with 3 doses of oral poliovirus vaccine (OPV) among 12-month-old children from 67% to 87%. We did a case-control study (70 case-patients, 692 age-matched controls) to estimate the clinical efficacy of OPV, assessed the immunogenicity of OPV and extent of poliovirus spread by serology, retrospectively evaluated the cold chain and vaccine potency, and sought the origin of the outbreak strain by genomic sequencing.

Immunogenicity of tetravalent rhesus rotavirus vaccine administered with buffer and oral polio vaccine.

Between January and November 1989, we studied 174 infants aged 6 to 16 weeks in a randomized clinical trial to (1) determine the immunogenicity of a single dose of tetravalent rhesus rotavirus vaccine (RRV-TV) when administered with three different buffer regimens: no antacid buffer and small-volume (2.5-mL) and large-volume (30-mL) antacid buffer; and (2) examine the potential interference of RRV-TV on the immune response to oral polio vaccine. Immunogenicity of RRV-TV, measured as a fourfold rise in antibody titers to rotavirus, was similar in the groups receiving small- and large-dose buffer (45% and 49%, respectively) and significantly less in the group that received RRV-TV alone (23%).

Detection and identification of vaccine-related polioviruses by the polymerase chain reaction.

We have used the polymerase chain reaction (PCR) to obtain sensitive detection and identification of poliovirus RNA genomes. Primer pairs were designed to permit identification of each Sabin poliovaccine strain by the electrophoretic mobilities of the amplified DNA products (Sabin 1: 97 bp; Sabin 2: 71 bp; Sabin 3: 44 bp). The compositions of samples containing mixtures of vaccine strains could be readily determined by PCR.

Oligonucleotide probes for the specific detection of the wild poliovirus types 1 and 3 endemic to Brazil.

Synthetic oligodeoxynucleotide probes, 21-23 nucleotides in length, were prepared which specifically hybridize to the genomes of the wild type 1 and 3 polioviruses currently endemic to the northeastern region of Brazil. The probes are complementary to sequences near the 5'-terminus of the VP1 gene that differ substantially among genetically distant polioviruses but are largely conserved among related isolates. The probes have been routinely used in the laboratory surveillance of poliomyelitis cases in Brazil, permitting direct, rapid identification of the indigenous wild polioviruses by dot-blot hybridization.

Molecular approaches to enteroviral diagnosis in idiopathic cardiomyopathy and myocarditis.

Enteroviruses are thought to be etiologic agents in some cases of human myocarditis and dilated cardiomyopathy. Murine models of acute coxsackievirus B3 myocarditis implicate coxsackie B viruses as possible causes of human myocarditis. Indirect evidence implicating enteroviruses as causative agents in human heart disease derives from serologic studies.

Echovirus 22 is an atypical enterovirus.

Although echovirus 22 (EV22) is classified as an enterovirus in the family Picornaviridae, it is atypical of the enterovirus paradigm, typified by the polioviruses and the coxsackie B viruses. cDNA reverse transcribed from coxsackievirus B3 (CVB3) RNA does not hybridize to genomic RNA of EV22, and conversely, cDNA made to EV22 does not hybridize to CVB3 genomic RNA or to molecular clones of CVB3 or poliovirus type 1. EV22 cDNA does not hybridize to viral RNA of encephalomyocarditis virus or to a molecular clone of Theiler's murine encephalomyelitis virus, members of the cardiovirus genus.

A molecular and serologic evaluation of enteroviral involvement in human myocarditis.

Murine coxsackie B virus infection models of myocarditis and numerous human serologic studies associating elevated enterovirus-specific IgM titers with the clinical diagnosis of myocarditis have been used to support an etiologic role for enteroviruses in human myocarditis. Of the human enteroviruses, coxsackie B viruses (CVB) are the enterovirus group most commonly associated with the human disease. While hybridization probes exist to detect most, if not all, human enteroviruses, no probe capable of specifically detecting an enteroviral group (such as the CVB) to the exclusion of all others has been described to date.