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At least 19 recordsLinked to original sources
PubMed1994

Detection of immunoglobulin G to measles virus, rubella virus, and mumps virus in serum samples and in microquantities of whole blood dried on filter paper.

Immunity to measles virus, rubella virus, and mumps virus was determined by EIA in serum samples and in dried whole blood specimens spotted on Whatman filter paper (5 mm in diameter). Both specimens were obtained from each patient by venepuncture and finger prick. Ten microliters of whole blood is enough to detect antibodies to these three different viruses. The comparison of the results obtained by EIA from 227 serum and whole blood samples have demonstrated close agreement: 98.6% for measles virus, 99.1% for rubella virus, and 96.0% for mumps virus. Moreover, 96 whole blood samples can be tested in a microtiter plate and can be stored at room temperature for 15 days or at +4 degrees C for several months. Therefore, whole blood dried on filter paper is a convenient alternative method for collecting and transporting specimens, it is easier and safer than venepuncture, and could be used for large-scale epidemiological studies, especially in newborns. This method could solve the problem of sampling, especially in young children, and could simplify studies of vaccine efficacy.

Adult↗
PubMed2002

Simultaneous detection of measles virus, rubella virus, and parvovirus B19 by using multiplex PCR.

We describe here a multiplex reverse transcription-PCR (RTMNPCR) assay designed to detect and differentiate measles virus, rubella virus, and parvovirus B19. Serial dilution experiments with vaccine strains that compared cell culture isolation of measles in B95 cells and rubella in RK13 cells showed sensitivity rates of 0.004 50% tissue culture infective dose (TCID(50)) for measles virus and 0.04 TCID(50) for rubella virus. This RTMNPCR can detect as few as 10 molecules for measles virus and rubella virus and one molecule for parvovirus B19 in dilution experiments with plasmids containing inserts of the primary reaction amplification products. Five pharyngeal exudates from measles patients and 2 of 15 cerebrospinal fluid samples from measles-related encephalitis were found to be positive for measles virus by this RTMNPCR. A total of 3 of 27 pharyngeal exudates from vaccinated children and 2 pharyngeal exudates, plus one urine sample from a case of congenital rubella syndrome, were found to be positive for rubella virus by RTMNPCR, whereas 16 of 19 sera from patients with erythema infectiosum were determined to be positive for parvovirus B19 by RTMNPCR. In view of these results, we can assess that this method is a useful tool in the diagnosis of these three viruses and could be used as an effective surveillance tool in measles eradication programs.

Child↗
PubMed1992

Virus-specific antibodies to Epstein-Barr virus, varicella-zoster virus and rubella virus in renal transplant patients with cytomegalovirus infections.

Renal transplant patients with primary and recurrent cytomegalovirus (CMV) infection had higher antibody titres to Epstein-Barr virus viral capsid antigen (EBV-VCA-IgG) before and after transplantation than healthy blood donors. The geometric mean titres (GMT) of EBV-VCA-IgG were higher in renal transplant patients without CMV infection than in renal transplant patients with CMV infection. Four-fold or greater rises in EBV-VCA-IgG antibody were detected in six patients and a similar rise in antibody to EBV early antigen (EBV-EA-IgG) was detected in one other patient. IgM antibody to EBV-VCA (EBV-VCA-IgM) was detected in only three of these patients. EBV-EA-IgG was present in 39% patients and in 30% control subjects. IgG titres to varicella zoster virus (VZV-IgG) and rubella virus (rubella HI) were higher in patients without CMV infection compared to the patients with CMV infection. Raised titres were detected to VZV in five patients and to rubella virus in three patients. Reductions in antibody titre of four-fold or more were also detected in EBV-EA-IgG (one patient) and to rubella virus (one patient). Raised antibody titres to EBV, VZV, and rubella virus in renal transplant patients may indicate reactivation of these viruses without any symptoms.

Antibodies, Viral↗
PubMed1968

Density gradient centrifugation of rubella virus.

Rubella virus was centrifuged in sucrose density gradients. One of two densities could be ascribed to the virus, depending upon the suspending medium used. The virus was found at a density of 1.16 g/cm(3) after centrifugation for 18 hr in sucrose gradients prepared in distilled water. By contrast, when the sucrose gradients were prepared in tris(hydroxymethyl)aminomethane (Tris)buffer containing ethylenediaminetetraacetic acid (EDTA), the virus was found at a density of 1.18 g/cm(3) after 18 hr of centrifugation. The virus banded at this higher density after only 2 hr of centrifugation when pretreated by overnight incubation in the Tris-EDTA buffer. A kinetic study showed that, in sucrose gradients containing this buffer, the virus gradually migrated as a single peak of infectivity from a density of 1.16 g/cm(3) after 2 hr of centrifugation to the higher 1.18 g/cm(3) density after 18 hr. The density change was shown to be reversible; after the removal of the Tris-EDTA buffer, rebanding of virus harvested at the heavy density resulted in its banding at the lower 1.16 g/cm(3) density. The data indicate that density change could not be explained on the basis of the loss of some component from the virus or on the basis of the failure of the virus to reach equilibrium. However, it is possible that the two densities observed were a reflection of the existence of rubella virus in different hydration states in the presence and absence of Tris buffer containing EDTA.

Centrifugation, Density Gradient↗
PubMed1972

Growth of Newcastle disease virus and rubella virus in rheumatoid and nonrehumatoid synovial cell cultures.

Rheumatoid and nonrheumatoid synovial cell cultures were challenged with Newcastle disease virus and rubella virus in an attempt to confirm reports that rheumatoid synovial cells are relatively resistant to infection with these viruses. Newcastle disease virus caused complete cell destruction by day 7 in both rheumatoid and nonrheumatoid cultures, and peak virus titers were similar. Rubella virus replicated in both rheumatoid and nonrheumatoid synovial cell cultures, and no consistent differences in virus titers were detected. Rubella-infected cell lines were observed for up to 28 days and no virus-specific cytopathic effect was seen.

Adolescent↗
PubMed1993

[Double meningoencephalitis with herpes simplex virus and rubella virus].

A 47-year-old man suffered from a headache, fever and memory disturbance. He was admitted to Morimoto Hospital. Neurological examination revealed disturbance of memory of recent events. Cranial nerves were normal. Muscle strength of the extremities was normal, except for Barré's sign of the right upper extremity. Deep tendon reflexes were exaggerated bilaterally, and extensor plantar response of the left side was elicited. Sensory examination showed no abnormality. Cerebellar sign was not recognized. Meningeal sign was slightly but clearly showed. CT scan demonstrated brain swelling at the right insular cortex region followed by severe hydrocephalus with dilatation of the lateral and the third ventricles. Cerebrospinal fluid showed high CSF pressure (250 mmH2O), pleocytosis (C.C. 359/mm3) and elevated protein level (213 mg/dl). Virological examination revealed herpes simplex virus (HSV) (CF) 32x, HSV-1 IgG (EIA) 4,050X, rubella virus IgE (EIA) 6,060X, cytomegalovirus (CMV) IgG (EIA) 1,130X in serum and HSV (CF) 1X, HSV-1 IGg (EIA) 1,430X, rubella virus IgG (EIA) 1,480X, CMV IgG (EIA) 587X in CSF. The ratio of serum/CSF of HSV and rubella virus titers by EIA methods were 2.83 and 4.10, respectively. He was treated by acyclovir 1,000 mg/day and gamma globulin, but his condition get worse acutely and died at 15th hospital days. This case was considered as a meningoencephalitis caused by simultaneous HSV and rubella virus infection.

Antibodies, Viral↗
PubMed1993

Dot immunobinding assay for simultaneous detection of specific immunoglobulin G antibodies to measles virus, mumps virus, and rubella virus.

A dot immunobinding assay was used to detect antibodies to measles virus, mumps virus, and rubella virus antigens. Filter paper soaked with serum or whole blood was directly applied to the antigen-coated nitrocellulose sheets. The test was easy to perform, and its results agreed very well with those obtained by standard enzyme immunoassay.

Antibodies, Viral↗
PubMedDate not supplied

[Comparison of the latex agglutination method with the hemagglutination inhibition test in the detection of antibodies against rubella virus].

Rubella virus antibodies were measured in 85 sera from pregnant women by using a new latex test, and the results were compared with those obtained by using hemagglutination inhibition. The sensitivity of the latex test was 98.4%, specificity was 66.6% and the predictive value of a positive result was 90%. The latex test is a simple test and has much shorter reaction time than that of the hemagglutination inhibition.

Antibodies, Viral↗
PubMedDate not supplied

[Fractionation of serum by gel filtration: contamination of IgM fractions by non-specific inhibitors of hemagglutination of the rubella virus].

Rubella virus hemagglutination non-specific inhibitors, in sera fractionated on Bio-Gel A 5 m, elute in IgM containing fractions. In addition, in heparin-MnCl2 treated sera, the same inhibitors display a different chromatographic behaviour on Bio-Gel A 5m, being eluted earlier than IgM. Therefore, in order to avoid false-positive results due to the presence of non-specific inhibitors in IgM containing fractions it is necessary to verify the disappearance of hemagglutination-inhibition activity after 2-ME reduction.

Blood Chemical Analysis↗
PubMed1991

Oligomerization of the structural proteins of rubella virus.

Rubella virus contains, in addition to its RNA genome, a nucleocapsid protein (C) and two membrane proteins (E2 and E1). We have studied the association of these proteins during viral assembly and when expressed from cDNA constructs. The C protein was found to dimerize very shortly after synthesis; this dimer became disulfide-linked in the virion. Formation of the dimer was independent of the presence of other RV proteins. The membrane glycoproteins formed an E2E1 heterodimer, a minor fraction of which was also found to be disulfide-linked in the virion. This heterodimer also formed when the two proteins were coexpressed from cloned cDNA. Formation of the heterodimer preceded the transport of E2 to the Golgi, as judged by modification of the protein by Golgi-located enzymes. In the absence of E2, the E1 protein was slowly converted to high molecular weight aggregates.

Capsid↗
PubMed2000

Infectious cDNA clone of the RA27/3 vaccine strain of Rubella virus.

Rubella virus (RUB), a small plus-strand RNA virus, is a significant human pathogen. The RA27/3 vaccine strain of RUB is one of the most successful live attenuated vaccines developed. In this article, we report the construction of an RA27/3 infectious clone, a complete cDNA copy of the RA27/3 genome that can be transcribed in vitro to generate infectious RNA molecules. Virus generated from such in vitro transcripts was phenotypically similar to RA27/3 virus. To investigate the attenuation of the RA27/3 strain, a series of chimeras was made by the insertion of different fragments of the RA27/3 genome into an infectious clone based on the Therien wild-type strain of RUB. Analysis of the resulting chimeric viruses revealed that the pattern of RA27/3 attenuation in cell culture is complex: attenuating elements in the RA27/3 genome were found in the 5' untranslated region (UTR), a region of the nonstructural proteins containing the protease motif and the capsid gene. Within the 5' UTR, the attenuation determinant was mapped to nt 7. Surprisingly, these analyses also revealed a potentiating mutation at nt 164 of the RA27/3 genome. Although this determinant was within the coding sequences of the nonstructural proteins, the encoded amino acid had no effect on cell culture phenotype and thus the determinant may operate at the level of RNA structure. In addition to investigation of the mechanisms of RA27/3 attenuation, the availability of the RA27/3 infectious clone offers the opportunity for strict genetic control over RUB vaccine manufacturing, for development of novel DNA-based vaccines against RUB, and for development of recombinant RUB vaccines that also target other diseases.

5' Untranslated Regions↗
PubMed1984

Interferon-induced 2-5A synthetase activity in human peripheral blood mononuclear cells after immunization with influenza virus and rubella virus vaccines.

The interferon-induced enzyme 2-5A synthetase can be a sensitive indicator of activation of the human interferon system during viral infection or interferon therapy. To determine the response of the human interferon system to viral antigens, the level of 2-5A synthetase activity was monitored in peripheral blood mononuclear cells of healthy adults before and after immunization with influenza or rubella virus vaccine. The influenza virus-vaccinated individuals demonstrated increases in enzyme activity on days 1 and 11 in vivo, whereas those vaccinated with rubella virus vaccine showed an increase only on day 11. The difference in the day 1 in vivo 2-5A synthetase response in the two vaccinated groups could be demonstrated by in vitro incubations of peripheral blood mononuclear cells isolated approximately 90 days postvaccination with the two vaccines. The day 11 increase of enzyme activity in the rubella virus group showed a positive correlation with an increase in serum antibody titer, suggesting activation of the interferon system during antibody production in vivo after human exposure to virus antigens. The demonstration of increased 2-5A synthetase activity at specific times postimmunization in this investigation indicates that the interferon system is involved in the human in vivo response to virus vaccination.

2',5'-Oligoadenylate Synthetase↗
PubMed2002

Rescue of rubella virus replication-defective mutants using vaccinia virus recombinant expressing rubella virus nonstructural proteins.

The genome of rubella virus (RV) is translated into a polyprotein precusor, p200, of the nonstructural proteins (NSPs). This is proteolytically processed by a viral-encoded protease into two mature products, p150 and p90. p150 contains sequence corresponding to the predicted methyltransferase and protease activities, while p90 has sequence for the proposed helicase and RNA-dependent RNA polymerase activities. Processing of p200 is essential for RV viral replication. RV NSPs are responsible for viral RNA replication, in which a full-length negative-strand RNA serves as the intermediate for the replication of positive-strand genomic RNA and the transcription of subgenomic RNA. Previously we demonstrated that p200 synthesizes negative- but not positive-strand RNA, and that cleavage products p150/p90 are required for efficient production of positive-strand RNA. To determine whether p150 or p90 alone or together is involved in positive-strand RNA synthesis, vaccinia virus recombinants expressing individual NSPs were constructed and characterized. These were used in in vivo rescue experiments to complement replication-defective mutants in virus replication. A protease-inactive mutant was rescued by p200 or p150 provided in trans by using vaccinia virus recombinants. Thus this protease can function in trans. Rescue of cleavage-defective mutant by either p200 alone, or p150 plus p90 but not by p150 or p90 alone suggests that p150 and p90 function together as a replication complex in positive-strand RNA synthesis.

Cell Line↗
PubMed1989

Immune responses to wild and vaccine rubella viruses after rubella vaccination.

Antibody responses to individual structural proteins (E1, E2, and C) of the M33 wild rubella virus and the RA 27/3 live attenuated rubella strain were assayed by immunoblotting in 11 girls, following immunization with RA 27/3 rubella vaccine. Serum samples were drawn before immunization and at 10 days, 1 month, 1 year, 2 years, and 3 years afterwards. All the subjects showed antibodies to E1 glycoprotein of both the virus strains up to three years after immunization, indicating the importance of E1 in immunity. Antibodies to E1 were always present when with neutralizing activity was observed. Antibodies to E2 protein of both the viruses and to the C protein of the M33 virus gradually disappeared with time in some samples, while antibodies to C protein of the RA 27/3 virus strain were found persistently in all the sera.

Adolescent↗
PubMed1999

Mutational analysis, using a full-length rubella virus cDNA clone, of rubella virus E1 transmembrane and cytoplasmic domains required for virus release.

We report on the construction of a full-length cDNA clone, pBRM33, derived from wild-type rubella virus M33 strain. The RNA transcripts synthesized in vitro from pBRM33 are highly infectious, and the viruses produced retain the phenotypic characteristics of the parental M33 virus in growth rate and plaque size. This cDNA clone was used to study the role of E1 transmembrane and cytoplasmic domains in virus assembly by site-directed mutagenesis. Three different alanine substitutions were introduced in the transmembrane domain of E1. These included substitution of leucine 464, cysteine 466, cysteine 467, and both cysteines 466 and 467 to alanine. In the E1 cytoplasmic domain, cysteine 470 and leucine 471 were altered to alanine. We found that these mutations did not significantly affect viral RNA replication, viral structural protein synthesis and transport, or E2/E1 heterodimer formation. Except for the substitution of cysteine 470, these mutations did, however, lead to a reduction in virus release. Substitution of cysteine 467 in the transmembrane region and of leucine 471 in the cytoplasmic domain dramatically reduced virus yield, resulting in the production of only 1 and 10% of the parental virus yield, respectively, in a parallel infection. These data show that E1 transmembrane and cytoplasmic domains play an important role in late stages of virus assembly, possibly during virus budding, consistent with earlier studies indicating that the E1 cytoplasmic domain may interact with nucleocapsids and that this interaction drives virus budding.

Animals↗
PubMed1989

Examination of eight cases of multiple sclerosis and 56 neurological and non-neurological controls for genomic sequences of measles virus, canine distemper virus, simian virus 5 and rubella virus.

In situ hybridization studies have been carried out on brain samples from eight cases of multiple sclerosis (MS) and 56 non-neurological and neurological controls, using single-stranded 35S-labelled RNA probes prepared against genomic RNA sequences of measles virus, canine distemper virus, rubella virus and simian virus 5. Foci of hybridization were found using probes against the measles virus nucleocapsid protein (N), phosphoprotein and fusion protein gene sequences in two of the MS cases, and also in one control, a case of disseminated cytomegalovirus infection with spinal cord necrosis. This result was confirmed using biotinylated probes prepared against the measles virus N genomic sequence. No hybridization was found in any of the MS or control cases using any of the other viral genome-specific probes.

Astrocytoma↗
PubMed1974

Impaired cellular immunity to rubella virus in congenital rubella.

Specific cell-mediated immunity (CMI) responses to rubella virus were studied in 12 children with documented congenital rubella syndrome employing a (51)Cr lymphocytotoxicity microassay. Hemagglutination inhibition antibody was detected in 11 of the 12 children, with titers ranging from 1:4 to 1:128. CMI to rubella virus was demonstrated in only 3 of the 11 antibody-positive children. The 12th child was negative for both hemagglutination inhibition and CMI. Of the three children with a positive CMI response, two had histories of reinfection with rubella virus. These data suggest that congenital rubella infection produces an impaired CMI response which subsequently may be altered by reinfection with rubella virus. The lack of CMI in the presence of antibody and concurrent excretion of live virus in the child with documented congenital rubella infection suggest a factor to be explored in the pathogenesis of this disease.

Adolescent↗