RESUMO
Immunity to rubella virus (RV) is conventionally determined by measuring specific immunoglobulin G (IgG). However, several individuals may be considered immune despite undetectable antibody levels. In the present study RV-specific interferon-gamma (IFN gamma)-ELISpot and rubella-IgG-ELISA were compared in 75 young adults aged between 20 and 30 years. In a subgroup, not only rubella-like particles (RLP), but also HPV77 rubella vaccine derived antigen was used in IFN gamma-ELISpot. The results from both, ELISA and ELISpot were independent of previous encounter to RV (vaccination, exanthematous disease, or childhood infection). There was no difference between RLP and RV vaccine antigen in IFN gamma-ELISpot response, and there was no correlation between IFN gamma-ELISpot and RV-specific IgG levels. IFN gamma-producing cells were found in 78.7% of all tested persons, and 83.8% of them were positive in ELISA. In almost all individuals seronegative for RV antibody, IFN gamma-producing cells were detected. Considering both humoral and cell-mediated immune responses, a positive RV immune reaction was seen in 98.6%. The results indicate that the IFN gamma-ELISpot can provide valuable additional information in seronegative individuals.
Assuntos
Imunidade Celular , Imunoensaio/métodos , Interferon gama/metabolismo , Leucócitos Mononucleares/imunologia , Vírus da Rubéola/imunologia , Adulto , Anticorpos Antivirais/sangue , Antígenos Virais , Células Cultivadas , Feminino , Humanos , Imunidade Humoral , Imunoglobulina G/sangue , Masculino , Virossomos , Adulto JovemRESUMO
31 prematures with signs of the cytomegalovirus infection (CMV) were examined. The blood and urine samples were tested for direct viral markers, i.e. for infectious CMV by the rapid culture method (RCM) and for viral DNA by quantitative PCR. Besides, the parameters of the specific immune response were studied in the babies. CMV was detected by RCM and/or PCR in 25 of the 31 examined babies during their 1st life week. The highest content of CMV within the investigated samples, i.e. 100 antigen-containing cells per 2.5 x 10(5) culture cells and above 2000 copies/ml of viral DNA was detected in 8 (32%) children. The quantity of viral DNA did not exceed 1000 copies/ml and one to three of stained cells was detected by PCR in 13 (42%) children. A study of anti-CMV in sera revealed high-titer of AT IgG in all 30 children. High avidity of anti-CMV-IgG was demonstrated to correlate with a low viral load and a low CMV infection activity in the newborns. According to the results, at least 3 laboratory diagnosis tools should be used in the diagnosis, they are PCR, RCM and determination of the anti-CMV avidity.
Assuntos
Infecções por Citomegalovirus/diagnóstico , Citomegalovirus/isolamento & purificação , Recém-Nascido Prematuro , Anticorpos Antivirais/sangue , Afinidade de Anticorpos , Antígenos Virais/sangue , Antígenos Virais/urina , Células Cultivadas , Citomegalovirus/genética , Citomegalovirus/imunologia , Infecções por Citomegalovirus/sangue , Infecções por Citomegalovirus/urina , DNA Viral/sangue , DNA Viral/urina , Humanos , Imunoglobulina G/sangue , Recém-Nascido , Recém-Nascido Prematuro/sangue , Recém-Nascido Prematuro/urina , Reação em Cadeia da Polimerase , Cultura de VírusRESUMO
Thirty-three children aged 1 month to 3 years were examined within the case study. spELISA, immunoblot (IB), shell vial method (SVM) and PCR, were used for the detection of anti-CMV IgM and IgG, in the diagnosis of cytomegalovirus (CMV). Clinical signs of CMV infection (CMVI) were registered in 20 children (group 1); no CMVI specific signs were detected in the remaining 13 children (group 2). Class M antibodies were identified in 50% of group-1 sera. Around 80% of children in the group had anti-CMV-IgG. AI < 0.6 was in 3 (20%) of 15 examinees. Direct CMV markers (DNA and infection activity) were detected in 13 (65%) of 20 children. Sera of 13 children with non-specific symptomatology (group 2) had no anti-CNV-IgM, while IgG were found in 54% examinees in the group. The infectious active virus was not detected in a single baby. The used laboratory tools enhance the efficiency of CMVI diagnosis and denote a disease variation.
Assuntos
Infecções por Citomegalovirus/diagnóstico , Citomegalovirus/isolamento & purificação , Anticorpos Antivirais/sangue , Afinidade de Anticorpos , Pré-Escolar , Citomegalovirus/genética , Citomegalovirus/imunologia , Infecções por Citomegalovirus/sangue , Infecções por Citomegalovirus/virologia , DNA Viral/análise , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Immunoblotting , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Lactente , Recém-Nascido , Reação em Cadeia da Polimerase , Cultura de VírusRESUMO
A total of 44 serum specimens from 7 patients with kidneys or liver transplanted from donors who had antibodies (Ab) to human cytomegalovirus (CMV) were studied. In 4 recipients anti-CMV Ab were found before transplantation and in 3 others they were not detected. It was shown by EIA that IgM and IgG anti-CMV appeared in the sera of primarily infected patients after 1-2 weeks and their titers were 5-10 times lower than in patients with reactivated CMV infection. Immunoblotting of Ab to individual CMV proteins showed a narrower spectrum of Ab during the initial period of primary CMV infection in comparison with the same period of reactivation and delayed production of AB to conformation-dependent determinants. Hence, analysis of anti-CMV Ab during the first 4-6 weeks after organ transplantation by EIA and immunoblotting differentiates primary CMV infection from its reactivation by Ab titers and spectrum. These parameters vary in some patients.
Assuntos
Anticorpos Antivirais/imunologia , Infecções por Citomegalovirus/imunologia , Citomegalovirus/imunologia , Antígenos Virais/química , Antígenos Virais/imunologia , Infecções por Citomegalovirus/etiologia , Humanos , Imunidade , Epitopos Imunodominantes , Infecções Oportunistas/imunologia , Transplante de Órgãos/efeitos adversos , Conformação Proteica , Relação Estrutura-Atividade , Imunologia de Transplantes , Transplante HomólogoRESUMO
A common problem in rheumatological practice is inflammatory joint disease that cannot be classified. The prognosis of such undifferentiated arthritides is uncertain. The synovial tissue of 41 consecutive patients with various forms of arthritis was tested for the presence of viral DNA in a diagnosis-unaware fashion, using the polymerase chain reaction (PCR). Of all tested viruses, cytomegalovirus and parvovirus B19 were positive (each in 10 patients, two double-positives), whereas herpes simplex virus was positive in two patients. Rubella virus RNA was detected in three specimens. When the positivity for viral material was analysed in terms of distribution among the various diagnostic groups, it became evident that five out of 10 parvovirus B 19-positive patients belonged to the undifferentiated arthritis group, whereas cytomegalovirus-positive patients were spread among all diagnostic groups. This indicates the possibility of a new diagnostic category of undifferentiated mono- and oligoarthritis, which can be identified by the presence of parvovirus B19 DNA in synovial tissue.
Assuntos
Artrite/virologia , DNA Viral/análise , Parvovirus/genética , Membrana Sinovial/virologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Anticorpos Antivirais/análise , Artrite/classificação , Artrite/imunologia , Sequência de Bases , Grupo Borrelia Burgdorferi , Criança , Chlorocebus aethiops , Citomegalovirus/genética , Citomegalovirus/imunologia , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Células HeLa/virologia , Humanos , Incidência , Doença de Lyme , Pessoa de Meia-Idade , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , RNA Viral/análise , Vírus da Rubéola/genética , Vírus da Rubéola/imunologia , Simplexvirus/genética , Simplexvirus/imunologia , Células Vero/virologiaRESUMO
Inadvertent immunisation of seronegative women with RA27/3 rubella virus live-attenuated vaccine several weeks before and after conception is described. Whereas in 5 cases the vaccine virus was not transmitted vertically, in 1 case vaccination led to the development of persistent fetal infection with prolonged virus shedding for more than 8 months. Sequence analysis carried out on isolates from amniotic fluid, from cord blood leukocytes as well as from infantile urine confirmed an infection by the vaccine strain. At birth, the newborn infant exhibited none of the symptoms compatible with the congenital rubella syndrome and signs indicative for development of late onset disease are not apparent. This observation constitutes the first unequivocal documented case of rubella vaccine virus related to persistent fetal infection.
Assuntos
Doenças Fetais/virologia , Complicações Infecciosas na Gravidez/virologia , Vírus da Rubéola , Rubéola (Sarampo Alemão)/transmissão , Vacinação/efeitos adversos , Adulto , Animais , Chlorocebus aethiops , Feminino , Doenças Fetais/prevenção & controle , Humanos , Lactente , Masculino , Gravidez , Complicações Infecciosas na Gravidez/prevenção & controle , RNA Viral/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rubéola (Sarampo Alemão)/congênito , Rubéola (Sarampo Alemão)/prevenção & controle , Rubéola (Sarampo Alemão)/virologia , Vírus da Rubéola/genética , Vírus da Rubéola/isolamento & purificação , Análise de Sequência de DNA , Células VeroAssuntos
Infecções/microbiologia , Transtornos da Lactação/microbiologia , Complicações Infecciosas na Gravidez/microbiologia , Feminino , Humanos , Infecções/diagnóstico , Infecções/terapia , Transtornos da Lactação/diagnóstico , Transtornos da Lactação/terapia , Gravidez , Complicações Infecciosas na Gravidez/diagnóstico , Complicações Infecciosas na Gravidez/terapiaAssuntos
Complicações Infecciosas na Gravidez/diagnóstico , Síndrome da Rubéola Congênita/diagnóstico , Rubéola (Sarampo Alemão)/diagnóstico , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Gravidez , Complicações Infecciosas na Gravidez/prevenção & controle , Diagnóstico Pré-Natal , Recidiva , Rubéola (Sarampo Alemão)/prevenção & controle , Síndrome da Rubéola Congênita/prevenção & controle , Vacina contra Rubéola/administração & dosagemRESUMO
A 4-year-old girl with condylomata acuminata of the vulva and papular warts of the surrounding skin is presented. The lesions were removed by surgery. Histologic investigation showed koilocytosis of the squamous epithelium and in-situ hybridization revealed human papilloma virus type 6 infection. There were no signs of sexual abuse or sexual transmission of the virus. After ablation, an interferon-containing ointment was applied. In order to prevent recurrence, a low-molecular-weight immunomodulating leucocyte fraction was given for more than 1 year, during which time no relapse was observed.
Assuntos
Condiloma Acuminado/terapia , Papillomaviridae , Doenças da Vulva/terapia , Adjuvantes Imunológicos/uso terapêutico , Antivirais/uso terapêutico , Pré-Escolar , Condiloma Acuminado/cirurgia , Condiloma Acuminado/virologia , Feminino , Humanos , Interferon beta , Doenças da Vulva/cirurgia , Doenças da Vulva/virologiaRESUMO
In an attempt to define diagnostic criteria that may help to distinguish the congenital rubella syndrome (CRS) from subclinical intrauterine rubella virus (RV) infection, maternal and fetal serum samples were analyzed using (1) enzyme immunoassay employing RV synthetic peptides as antigen, (2) IgG avidity assay, and (3) immunoblot under nonreducing conditions, in addition to hemagglutination inhibition and commercial enzyme immunoassays. Infants born with CRS and their mothers were shown to reveal low or undetectable levels of E2-specific antibodies and deficient IgG recognizing the major neutralizing antibody-inducing epitope on the E1 protein (SP15). Antibody responses were normal in mothers with presumed RV reinfection as well as in asymptomatic infants born after maternal primary rubella. The results indicate that the maturation of specific humoral immune responses is obviously less efficient when intrauterine RV infection results in CRS. The detection of high avidity IgG, conformational E2-specific as well as SP15-reactive antibodies may serve as a potential predictor for a benign outcome of intrauterine RV infections.
Assuntos
Anticorpos Antivirais/sangue , Complicações Infecciosas na Gravidez/diagnóstico , Síndrome da Rubéola Congênita/diagnóstico , Vírus da Rubéola/imunologia , Rubéola (Sarampo Alemão)/diagnóstico , Feminino , Sangue Fetal/virologia , Humanos , Recém-Nascido , Transmissão Vertical de Doenças Infecciosas , Valor Preditivo dos Testes , Gravidez , Complicações Infecciosas na Gravidez/virologia , Rubéola (Sarampo Alemão)/virologia , Síndrome da Rubéola Congênita/embriologia , Síndrome da Rubéola Congênita/virologia , Testes Sorológicos , Útero/virologiaRESUMO
Rubella infection during the first trimester of pregnancy can cause the congenital rubella syndrome (CRS). Patients with CRS were shown to have a decreased humoral and cellular immunity. It is not known whether asymptomatic newborns who had experienced intrauterine infection with rubella virus (RV) differ in their antibody response from newborns with CRS. In this study we compared both groups for a difference which might be a useful diagnostic criterion for CRS during the prenatal and newborn periods. We used the nonreducing Rubella-Immunoblot and the Rubella-IgG-Peptide-Enzyme Immunoassay (EIA) to determine the antibodies directed to rubella proteins E1, E2 and C. The results showed that only newborns with CRS who had experienced RV infection during the first 12 weeks of gestation showed significantly reduced levels of antibodies directed to both the linear RV E1 epitope (SP 15) and the topographic RV E2 epitope. Asymptomatic newborns infected mostly later than week 10 of gestation showed normal levels of antibodies. These data suggest that the lack of antibody response in CRS is linked to the immaturity of the fetal immune system during the first trimester of gestation. Rubella-IgG-Peptide-EIA and Rubella-Immunoblot should be used additionally for CRS diagnosis in the prenatal/newborn periods. These results may have an impact on the early treatment of late-onset symptoms of CRS patients.
Assuntos
Anticorpos Antivirais/análise , Immunoblotting , Técnicas Imunoenzimáticas , Síndrome da Rubéola Congênita/diagnóstico , Rubéola (Sarampo Alemão)/diagnóstico , Feminino , Humanos , Imunoglobulina G/análise , Recém-Nascido , Gravidez , Diagnóstico Pré-Natal , Rubéola (Sarampo Alemão)/imunologia , Síndrome da Rubéola Congênita/imunologia , Proteínas do Core Viral/imunologia , Proteínas do Envelope Viral/imunologiaRESUMO
BACKGROUND: We have investigated the performance of the novel rubella serology assay, Cobas Core Rubella IgG EIA recomb, which uses rubella-like particles (RLPs) expressed in transfected BHK-21 cells as the antigen. STUDY DESIGN: Evaluation of the assay included comparison with the hemagglutination inhibition (HAI) assay and another enzyme immunoassay (EIA) using native rubella virus (RV) as antigen, i.e. the Abbott IMx Rubella IgG. The assay was calibrated against the WHO 1000 IU/ml reference serum and showed good correlation with the HAI test in the analysis of 404 serum samples. However, quantitative differences in IgG values measured in the Cobas Core and the Abbott IMx assays were noted. RESULTS: Values obtained for patient sera as well as CDC and WHO standards were generally more than twice as high in the Abbott IMx assay as in the Cobas Core test. CONCLUSIONS: For sera whose IgG levels in the immunoassay and HAI test were discordant, immunoblotting proved valuable as a confirmatory method and indicated that a significant number of HAI-negative samples were correctly interpreted as positive by the immunoassay.
RESUMO
BACKGROUND: We investigated the utility of cytologic studies (including follow-up studies) in our specialty in determining the prevalence of human papillomavirus, specifically in the larynx. We utilized Papanicolaou's method of exfoliative cytology and in situ hybridization of the culture (biotin-marked DNA probes). PATIENTS: 486 patients with chronic hyperplastic laryngitis and 74 patients with papilloma were enrolled in the study. RESULTS: In 198 typings in 132 patients (59 patients with papilloma, 41 with chronic hyperplastic laryngitis, 17 with cancer, and 15 healthy), we found distributions of the individual types that were independent of the diagnosis. Particularly often, we encountered human papillomavirus types 18 and 31 in the presence of dysplasia, human papillomavirus type 16 in the presence of cancer, and type 11 in general. CONCLUSIONS: Cytologic examination and human papillomavirus typing of the culture permit us to define a third risk category in addition to the known risk groups smokers and dysplasia patients. Besides this, cytologic follow-up studies can provide further insight into the behavior of preneoplastic epithelial changes.
Assuntos
Neoplasias Laríngeas/patologia , Laringite/patologia , Papiloma/patologia , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/patologia , Lesões Pré-Cancerosas/patologia , Infecções Tumorais por Vírus/patologia , Adulto , Idoso , Biópsia , Transformação Celular Neoplásica/patologia , Doença Crônica , Feminino , Seguimentos , Humanos , Hiperplasia , Neoplasias Laríngeas/virologia , Laringite/virologia , Laringe/patologia , Laringe/virologia , Masculino , Pessoa de Meia-Idade , Papiloma/virologia , Infecções por Papillomavirus/virologia , Lesões Pré-Cancerosas/virologia , Infecções Tumorais por Vírus/virologiaRESUMO
Twenty-six mouse hybridomas producing monoclonal antibodies (MAB) to human cytomegalovirus (CMV) proteins have been obtained. MAB produced by three hybridomas were studied in detail. MAB were active in indirect immunofluorescence and solid-phase enzyme immunoassay, being directed to the super-early viral protein p72. Use of the resultant MAB for analysis of CMV-infected cells demonstrated that by specificity and sensitivity of viral antigen detection they were not inferior to anti-CMV antibodies manufactured by Ortho, USA. Screening of 258 patients with suspected CMV infection showed that these MAB may be used for immunofluorescent detection of CMV antigens in the material isolated from patients and infected subjects.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos Virais/imunologia , Citomegalovirus/isolamento & purificação , Proteínas Imediatamente Precoces/imunologia , Animais , Anticorpos Monoclonais/biossíntese , Células Cultivadas , Citomegalovirus/imunologia , Infecções por Citomegalovirus/diagnóstico , Imunofluorescência , Humanos , Hibridomas , Técnicas Imunoenzimáticas , Camundongos , Sensibilidade e EspecificidadeRESUMO
Cobas Core Rubella IgG EIA recomb (Roche), a new, commercially available rubella enzyme immunoassay using recombinant rubella-like particles, was compared with a hemagglutination inhibition assay (Rubenosticon; Organon Teknika) and two whole-virus enzyme immunoassays (IMx [Abbott] and Platelia [Sanofi Diagnostics Pasteur]). Compared with those of these reference assays, the relative sensitivities of Cobas Core Rubella IgG recomb were 100, 94, and 95.9%, with specificities amounting to 80.8, 98, and 98.2%, respectively.
Assuntos
Anticorpos Antivirais/sangue , Técnicas Imunoenzimáticas , Vírus da Rubéola/imunologia , Rubéola (Sarampo Alemão)/imunologia , Humanos , Proteínas Recombinantes/imunologia , Rubéola (Sarampo Alemão)/diagnóstico , Proteínas Virais/imunologiaRESUMO
In 8 out of 29 patients with scleroderma we found antibodies to HIV retroviral proteins in the Western blot analysis. The sera each reacted only to one or two of the p 18, p 24, p 55, and p 65 bands, and the reactions were relatively weak. There were no evident clinical correlations with the reactivity of certain bands and signs of direct HIV infection in our patients. Apart from three cases with positive CMV reactivity (IgM) there was no cross-reactivity to HTLV I or EBV (IgM) or to topoisomerase (Scl 70) or other autoantibodies to various nuclear antigens related to scleroderma. It is not clear whether retroviruses are involved into the pathogenesis of scleroderma or whether these antibodies are due to molecular mimicry.
Assuntos
Anticorpos Antivirais/análise , Anticorpos Anti-HIV/análise , Soropositividade para HIV/imunologia , HIV-1/imunologia , Proteínas dos Retroviridae/imunologia , Escleroderma Sistêmico/imunologia , Autoanticorpos/análise , Western Blotting , Dermatite Atópica/diagnóstico , Dermatite Atópica/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Soropositividade para HIV/diagnóstico , Humanos , Imunoglobulina G/análise , Masculino , Pessoa de Meia-Idade , Psoríase/diagnóstico , Psoríase/imunologia , Escleroderma Sistêmico/diagnósticoRESUMO
In 8 out of 29 patients with scleroderma we found antibodies to HIV retroviral proteins in the Western blot analysis. The sera reacted only to one or two of the following bands: p 18, p 24, p 55, p 65 in relatively weak grades. There were no evident clinical correlations with the reactivity of certain bands, nor signs of direct HIV infection in our patients. Apart from 3 cases with positive CMV reactivity (IgM), there was no cross-reactivity to HTLV I or EBV (IgM) and to topoisomerase (Scl 70) or other autoantibodies to various nuclear antigens related to scleroderma. It is not clear whether retroviruses are involved in the pathogenesis of scleroderma or whether these antibodies are due to molecular mimicry.
Assuntos
Anticorpos Antivirais/sangue , HIV-1 , Proteínas dos Retroviridae/imunologia , Escleroderma Sistêmico/sangue , Adulto , Anticorpos Antinucleares/sangue , Autoanticorpos/sangue , Reações Cruzadas , Citomegalovirus/imunologia , DNA Topoisomerases Tipo I , Feminino , Anticorpos Anti-HIV/sangue , Anticorpos Anti-HTLV-I/sangue , Herpesvirus Humano 4/imunologia , Humanos , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Proteínas Nucleares/sangueRESUMO
For diagnosis of the interstitial pneumonia conditioned by cytomegalovirus (CMV) an indirect immunofluorescence test was used to measure IgM and IgG antibodies to CMV in 369 serum specimens from 41 patients after bone marrow transplantation (BMT). An interstitial pneumonia conditioned by CMV was diagnosed in 5 patients either serological (4 patients) or by detection of cytomegaloviral infection typical inclusion bodies in lung cells (1 patient). The detection of antibodies to CMV for diagnosis of interstitial pneumonia conditioned by CMV is problematical in the phase immediately after BMT because bone marrow recipients are severely immunosuppressed by radiation and cytotoxic drugs received before transplantation. New method for detection of CMV-specific antigens are necessary.
Assuntos
Anticorpos Antivirais/isolamento & purificação , Transplante de Medula Óssea , Infecções por Citomegalovirus/diagnóstico , Citomegalovirus/imunologia , Fibrose Pulmonar/microbiologia , Adolescente , Adulto , Transplante de Medula Óssea/imunologia , Infecções por Citomegalovirus/imunologia , Feminino , Imunofluorescência , Humanos , Hospedeiro Imunocomprometido , Masculino , Fibrose Pulmonar/imunologia , Estudos RetrospectivosRESUMO
A new commercial ELISA test kit "Rubella-IgG-EIA SSW" is described for the determination of IgG antibodies to rubella virus. A panel of 99 sera was tested by "Rubella-IgG-EIA SSW" and by hemagglutination-inhibition (HI test). The results of enzyme-immunoassay (EIA) and of HI test correlate well; the coefficient of correlation is 0.95, the specificity is 90.9% and the sensitivity is 100%. A coefficient of correlation was found out of 0.92 between EIA and HI test in the evaluation of the quantitative procedure. The test kit can be used for determination of immune status to rubella virus and for quantitative detection of rubella-IgG antibodies.