RESUMO
Human immunodeficiency virus (HIV) mother-to-child transmission is a complex event, depending upon environmental factors and is affected by host genetic factors from mother and child, as well as viral genetic elements. The integration of multiple parameters (CD4 cell count, virus load, HIV subtype, and host genetic markers) could account for the susceptibility to HIV infection, a multifactorial trait. The goal of this manuscript is to analyze the immunogenetic factors associated to HIV mother-to-child transmission, trying to unravel the genetic puzzle of HIV mother-to-child transmission and considering the experience in this topic of two research groups from Brazil and Argentina.
Assuntos
Infecções por HIV , Transmissão Vertical de Doenças Infecciosas , Complicações Infecciosas na Gravidez , Argentina , Brasil , Suscetibilidade a Doenças , Feminino , Infecções por HIV/genética , Infecções por HIV/imunologia , Infecções por HIV/transmissão , Infecções por HIV/virologia , Humanos , Lactente , Recém-Nascido , GravidezRESUMO
Human papillomavirus (HPV) has been extensively studied concerning genomic structure, infection mechanisms, and diversity of types, as well as disease progression stages and development of vaccines. HPV type prevalence can differ in specific populations in different countries, according to ethnicity. This is the first report of an integrated project to evaluate the incidence of HPV types in different regions in Brazil in order to obtain data for vaccine development. Cervical samples were collected from women seen at a public hospital in Pernambuco, Northeast Brazil, for routine evaluation of genital alterations. Selection of the patients was random. There was a strong prevalence of HPV16 and a high incidence of HPV types 31 and 33. These data foster the discussion about the need to evaluate viral prevalence in each geographic region in order to develop targeted vaccine programs.
Assuntos
Alphapapillomavirus/isolamento & purificação , Colo do Útero/virologia , Alphapapillomavirus/classificação , Alphapapillomavirus/genética , Brasil , Feminino , Humanos , Reação em Cadeia da Polimerase , Especificidade da EspécieRESUMO
BACKGROUND: Anti-transglutaminase antibodies are highly predictive markers of active coeliac disease. Because limited facilities are available for routine use of anti-transglutaminase antibodies assays in developing countries, a simple, economical immunological test would represent a great step forward in the screening of coeliac disease. AIM: We determined the prevalence of coeliac disease in two different populations living in an urban area and in a sub-urban impoverished area of Recife (Brazil), using two rapid tests based on detection of anti-transglutaminase antibodies in serum and in one drop of whole blood. METHODS: Whole-blood and serum samples from 1074 individuals were analysed by the two rapid tests; 580 subjects were university students and 494 subjects were coming from sub-urban impoverished areas, characterized by the endemic presence of filariasis. The positive subjects were evaluated by anti-tranglutaminase enzyme linked immunosorbent assay (ELISA) assay, the coeliac disease-related HLA DQ2/8 and intestinal biopsy. RESULTS: Both rapid assays were positive in 25/1074 subjects, but only 9/25 (4/4 in urban areas, specificity 100%; 5/21 in poor areas, specificity 76%) were confirmed positive by ELISA assay. The nine subjects testing positive for HLA DQ2 and the intestinal biopsy showed the typical coeliac disease lesions (coeliac disease-prevalence: 0.84%, 9/1074); seven coeliacs were asymptomatic and two presented recurrent abdominal pain. CONCLUSIONS: The rapid assays were accurate in finding new coeliacs at a remarkably low cost. We are convinced that this new way of testing for coeliac disease can be successfully used by non-specialized personnel in daily practice in developing countries.
Assuntos
Anticorpos/análise , Doença Celíaca/diagnóstico , Programas de Rastreamento/métodos , Transglutaminases/imunologia , Adulto , Biópsia , Brasil/epidemiologia , Doença Celíaca/epidemiologia , Doença Celíaca/imunologia , Países em Desenvolvimento , Diagnóstico Diferencial , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Incidência , Masculino , Estudos Retrospectivos , População Rural , População UrbanaRESUMO
We report a fast (less than 3 h) and cost-effective melting temperature assay method for the detection of single-nucleotide polymorphisms in the MBL2 gene. The protocol, which is based on the Corbett Rotor Gene real time PCR platform and SYBR Green I chemistry, yielded, in the cohorts studied, sensitive (100%) and specific (100%) PCR amplification without the use of costly fluorophore-labeled probes or post-PCR manipulation. At the end of the PCR, the dissociation protocol included a slow heating from 60 degrees to 95 degrees C in 0.2 degrees C steps, with an 8-s interval between steps. Melting curve profiles were obtained using the dissociation software of the Rotor Gene-3000 apparatus. Samples were analyzed in duplicate and in different PCR runs to test the reproducibility of this technique. No supplementary data handling is required to determine the MBL2 genotype. MBL2 genotyping performed on a cohort of 164 HIV-1-positive Brazilian children and 150 healthy controls, matched for age and sex and ethnic origin, yielded reproducible results confirmed by direct sequencing of the amplicon performed in blind. The three MBL2 variants (Arg52Cys, Gly54Asp, Gly57Glu) were grouped together and called allele 0, while the combination of three wild-type alleles was called allele A. The frequency of the A/A homozygotes was significantly higher among healthy controls (0.68) than in HIV-infected children (0.55; P = 0.0234) and the frequency of MBL2 0/0 homozygotes was higher among HIV-1-infected children than healthy controls (P = 0.0296). The 0 allele was significantly more frequent among the 164 HIV-1-infected children (0.29) than among the 150 healthy controls (0.18; P = 0.0032). Our data confirm the association between the presence of the mutated MBL2 allele (allele 0) and HIV-1 infection in perinatally exposed children. Our results are in agreement with the literature data which indicate that the presence of the allele 0 confers a relative risk of 1.37 for HIV-1 infection through vertical transmission.
Assuntos
Infecções por HIV/genética , Lectina de Ligação a Manose/genética , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Nucleotídeo Único , Estudos de Casos e Controles , Criança , Análise Custo-Benefício , Frequência do Gene , Infecções por HIV/transmissão , Humanos , Reação em Cadeia da Polimerase/economia , Reprodutibilidade dos Testes , TemperaturaRESUMO
We report a fast (less than 3 h) and cost-effective melting temperature assay method for the detection of single-nucleotide polymorphisms in the MBL2 gene. The protocol, which is based on the Corbett Rotor Gene real time PCR platform and SYBR Green I chemistry, yielded, in the cohorts studied, sensitive (100 percent) and specific (100 percent) PCR amplification without the use of costly fluorophore-labeled probes or post-PCR manipulation. At the end of the PCR, the dissociation protocol included a slow heating from 60° to 95°C in 0.2°C steps, with an 8-s interval between steps. Melting curve profiles were obtained using the dissociation software of the Rotor Gene-3000 apparatus. Samples were analyzed in duplicate and in different PCR runs to test the reproducibility of this technique. No supplementary data handling is required to determine the MBL2 genotype. MBL2 genotyping performed on a cohort of 164 HIV-1-positive Brazilian children and 150 healthy controls, matched for age and sex and ethnic origin, yielded reproducible results confirmed by direct sequencing of the amplicon performed in blind. The three MBL2 variants (Arg52Cys, Gly54Asp, Gly57Glu) were grouped together and called allele 0, while the combination of three wild-type alleles was called allele A. The frequency of the A/A homozygotes was significantly higher among healthy controls (0.68) than in HIV-infected children (0.55; P = 0.0234) and the frequency of MBL2 0/0 homozygotes was higher among HIV-1-infected children than healthy controls (P = 0.0296). The 0 allele was significantly more frequent among the 164 HIV-1-infected children (0.29) than among the 150 healthy controls (0.18; P = 0.0032). Our data confirm the association between the presence of the mutated MBL2 allele (allele 0) and HIV-1 infection in perinatally exposed children. Our results are in agreement with the literature data which indicate that the presence of the allele 0 confers a relative risk of 1.37 for HIV-1 infection through vertical transmission.
Assuntos
Criança , Humanos , Infecções por HIV/genética , Lectina de Ligação a Manose/genética , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase/métodos , Estudos de Casos e Controles , Análise Custo-Benefício , Frequência do Gene , Infecções por HIV/transmissão , Reação em Cadeia da Polimerase/economia , Reprodutibilidade dos Testes , TemperaturaRESUMO
The frequencies of four CCR5 promoter polymorphisms, and of the Delta32 deletion, have been evaluated in Brazilian HIV-1 positive (HIV+) and HIV-1 negative (HIV-) children, both born from HIV-1 positive mothers and healthy controls (HC), with the aim of investigating whether CCR5 polymorphisms could be associated to vertical transmission of HIV-1. One hundred and six HIV-1 positive children and 70 HIV-1 negative children were enrolled from impoverished areas of Recife (Brazil). We recruited also as healthy controls 104 uninfected children from the same ethnic background, matched for age and known to be not at risk for HIV-1 infection. CCR5 polymorphisms were detected by PCR amplification and direct sequencing. Although no significative divergence was found for CCR5 Delta32, CCR5-59356-C/T and CCR5-59653 C/T polymorphisms, the frequency of CCR5-59353-T/C and CCR5-59402-A/G genotypes differed among HIV+, HIV- and HC children. The presence of the CCR5-59353-TT genotype indicated a trend for increased risk of vertical transmission of HIV-1 infection in Brazilian children, while the presence of the CCR5-59402-AA genotype is suggestive for a protective effect against HIV-1 vertical transmission.
Assuntos
Infecções por HIV/genética , Infecções por HIV/transmissão , HIV-1 , Transmissão Vertical de Doenças Infecciosas , Polimorfismo Genético , Receptores CCR5/genética , Brasil , Criança , Pré-Escolar , Feminino , Frequência do Gene , Infecções por HIV/epidemiologia , Humanos , Masculino , Gravidez , Regiões Promotoras Genéticas/genéticaRESUMO
Fifty AIDS patients were studied. All patients had anti-HIV antibodies (ELISA) present and met OPAS/Caracas punctuation criteria for AIDS cases in adults. Cerebrospinal fluid (CSF) analysis included pressure, cytology (number of cytomorphological aspects), total protein and electrophoresis, glucose and chloride concentration. Bacteriological and mycological investigations were performed as well as agglutination tests for Cryptococcus. Complement fixation, indirect immunoflorescence, passive hemagglutination and/or ELISA tests were performed for syphilis, toxoplasmosis, viral and fungal infections. All CSF analysis were made in the same laboratory following the same methodology. CSF was altered in 45 cases (90.0%) of the 50 patients studied. The most important alterations observed were: gammaglobulin (55.5%) and total protein (51.1%) increase, hypercytosis (48.9%) and decrease of chloride concentration (40.0%). HIV antibodies were detected in 42 patients (93.3%). Toxomoplamosis, isolated or associated to other agents, was the most frequent opportunistic infection (57.7%). Cerebrospinal fluid should always be examined in AIDS patients with or without neurological symptoms.
Assuntos
Síndrome da Imunodeficiência Adquirida/líquido cefalorraquidiano , Infecções Oportunistas/líquido cefalorraquidiano , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valores de ReferênciaRESUMO
Foram estudados 50 pacientes com AIDS, todos estes pacientes apresentavam anticorpos ant-HIV1 (ELISA) e preenchiam os critérios de pontuaçao OPAS/Caracas de definiçao de casos de AIDS em adultos. A análise do liquido cefalorraqueano (LCR) incluiu: pressao; citologia (número de células e aspectos citomorfológicos); proteína total e eletroforese; concentraçoes de glicose, cloretos e testes imunológicos para sífilis, toxoplasmose e infecçoes virais (citomegalovírus, varicela-zoster, Herpes simplex, e HIV1). Investigaçoes bacteriológicas e micológicas (pesquisa direta e cultura), além de teste de aglutinaçao (látex) paracryptococcus foram também realizados. Os testes imunológicos usados foram fixaçao do complemento, imunofluorescência indireta, hemaglutinaçao passiva e/ou ELISA. Todos os LCR foram analisados no mesmo laboratório seguindo sempre a mesma metodologia. O LCR esteve alterado em 45 pacientes (90,0 por cento) dos 50 pacientes estudados. As principais alteraçoes encontradas no LCR foram: aumento de gamaglobulina em 25 casos (55,5 por cento); aumento da proteína total em 23 (51,1 por cento); hipercitose em 22 (48,9 por cento) e diminuiçao dos cloretos em 18(40,0 por cento). A detecçao de anticorpos anti- HIV1 estiveram presentes em 42 pacientes (93,3 por cento). Toxoplasmose isolada ou associada a outros agentes foi a infecçao oportunista mais frequente, detectada em 26 casos (57,7 por cento). O LCR deverá ser sempre analisado em todos os pacientes com AIDS, com ou sem sintomas neurológicos.