Infection à VIH-2 au Sénégal: échecs virologiques et résistances aux antirétroviraux (ARV)

Introduction: le VIH-2, endémique en Afrique de l'Ouest, est naturellement résistant aux inhibiteurs non nucléosidiques de la rétro transcriptase, ce qui rend difficile la prise en charge dans les pays en développement. Objectifs: déterminer la prévalence de l'échec virologique au 12éme et 24éme mois (M12 et M24) de traitement antirétroviral de première ligne chez les patients infectés par le VIH-2 et d'en décrire les résistances génotypiques associées.Méthodes: il s'agit d'une étude descriptive longitudinale et prospective, durant la période de Novembre 2005 à Juin 2017. L'échec virologique a été défini comme toute charge virale supérieure à 50 copies/ml après 6 mois de traitement ARV à deux reprises. La recherche de mutations de résistance a été réalisée dans les régions codantes de la protéase et de la transcriptase inverse. Résultats: au total 110 patients ont été colligés, d'âge médian de 46 ans (Extrêmes 18-67) avec un ratio F/H de 2,54. A l'inclusion, la charge virale était détectable dans 44% des cas avec une médiane de 935cp/ml (Extrêmes 17-144038). Le schéma antirétroviral associait 2 INTI à 1IP dans 94% des cas. La durée médiane de suivi était estimée à 1200 jours (Extrêmes 1-3840). 94 puis 76 patients ont respectivement complété leur bilan à M12 et M24. Au suivi M24, 39 patients étaient en échec virologique soit une prévalence de 39% estimée à 33% à M12 et 11% à M24. 45% des patients avaient des résistances aux INTI, 41% des résistances aux IP et 30% des multi résistances aux INTI et IP.Conclusion: il est impératif de rendre accessibles les nouvelles classes thérapeutiques pourle traitement de sauvetage des patients infectés par le VIH-2 dans les pays à ressources limitées

Inconclusive results with HIV serodiagnosis algorithms, and HIV-1 and HIV-2 co-infection in North-eastern Democratic Republic of Congo

Background:Inconclusive serodiagnosis of HIV infection is particularly frequent in Central Africa. The aims of this study were to: (i) determine the rate of inconclusive results with the two-test algorithm that the WHO proposed in 1997 (WHO II) versus the three-test algorithm (revised in 2012 and consolidated in 2015 by WHO) for HIV testing, and (ii) determine the prevalence of HIV-1 and HIV-2 co-infection in the north-eastern region of the Democratic Republic of the Congo (DRC).Methods:A multicentre cross-sectional study was performed between March and June 2016 in Kisangani and Bunia, the capital cities of Tshopo and Ituri provinces respectively. Alere Determine HIV-1/2 (Alere Medical Co. Ltd., Japan), Uni-GoldTM HIV (Trinity Biotech Manufacturing Ltd., Ireland) and recomLine HIV-1 and HIV-2 IgG (Biosynex, France) were the first, second and third tests in the serial algorithm.Results : The rate of inconclusive results was 1.1% (95% CI: 0.4 to 3.1) with the two-test algorithm and 0.4% (95% CI: 0.1 to 2.1) with the three-test algorithm (p less than 0.001). The prevalence of HIV-1 and HIV-2 co-infection among HIV positive sera was 16.7% (95% CI: 4.7 to 44.8).Conclusion:The three-test algorithm HIV testing strategy significantly reduces the rate of inconclusive results. In addition, the prevalence of HIV-1 and HIV-2 co-infection is higher in a context where HIV-2 infection is poorly documented. Large-scale research is essential to clarify these results

Evaluación de la estabilidad del sistema de diagnóstico inmunoenzimático DAVIH VIH-2 / Evaluation of the stability of the immunoenzymatic diagnostic system DAVIH VIH-2

Introducción: el estudio de la estabilidad de los componentes y el producto terminado constituye un importante requisito regulatorio en los diagnosticadores. Objetivo: realizar un estudio de estabilidad en tiempo real durante doce meses del sistema inmunoenzimático (ELISA) DAVIH VIH-2. Métodos: se realizó un estudio de estabilidad en tiempo real durante doce meses en tres lotes del diagnosticador DAVIH VIH-2, ELISA indirecto diseñado para la detección de anticuerpos contra el virus de inmunodeficiencia humana tipo 2 en suero o plasma humano. Se controlaron los requisitos de calidad de los componentes de acuerdo a sus especificaciones. Se estudió la normalidad de valores de densidad óptica/valor límite y la homogeneidad de las medias y varianzas mediante las dócimas de Grubbs y Cochran. Se estimó la precisión en los controles positivo y negativo del sistema y en seis muestras con diferente reactividad al virus de inmunodeficiencia humana tipo 2 mediante el cálculo del coeficiente de variación y se confeccionaron las cartas de control de los valores de las medias de densidad óptica respecto al tiempo. Resultados: los requisitos de calidad de cada componente se cumplieron durante 12 meses, excepto las características funcionales del conjugado a partir de los seis meses. Los valores en las dócimas de Grubbs y Cochran fueron menores que los valores críticos tabulados para α del 1 y 5 por ciento por lo que existió homogeneidad en las medias y las varianzas en todo el periodo. El coeficiente de variación se mantuvo inferior al 10 por ciento excepto en las muestras con reactividad media y baja, mientras que en las cartas de control, los valores de densidad óptica se mantuvieron en el límite de la media ±2 desviaciones estándar hasta el noveno mes(AU)

Epidemiologia molecular, evolução viral e perfil de resistência aos antirretrovirais em uma população de pacientes infectados por HIV-1 e/ou HIV-2 residentes em diferentes ilhas do arquipélago de Cabo Verde / Molecular epidemiology, viral evolution and profile of drug resistance mutations among HIV-1 and/or HIV-2 infected patients from distinct islands of the Cape Verde Archipelago

O diagnóstico sorológico da infecção pelo HIV-1 e HIV-2 teve início em Cabo Verde em 1987, mas pouco se sabe a respeito da diversidade genética desses vírus nessas ilhas, localizadas na costa Ocidental Africana. Neste estudo, caracterizamos a epidemiologia molecular do HIV-1 e HIV-2 em Cabo Verde, analisamos a origem dos principais clados de HIV introduzidos no país e descrevemos a ocorrência de mutações de resistência aos antirretrovirais (DRM) em indivíduos virgens de tratamento (ARTn) e pacientes em tratamento (ARTexp) oriundos das diferentes ilhas. Amostras de sangue, dados sociodemográfico e clínico-laboratoriais foram obtidos de 221 indivíduos HIV positivos entre 2010-2011. As amostras foram sequenciadas na região da polimerase (1300 pares de bases) e análises filogenéticas e de bootscan foram realizadas para a subtipagem viral. Os algoritmos disponibilizados nos sites Stanford HIV Database e HIV-GRADE e.V. Algorithm Homepage foram utilizados para avaliar a existência de DRM em pacientes positivos para HIV-1 e HIV-2, respectivamente. Os estudos evolutivos e filogeográficos foram realizados através do programa BEASTEntre os 221 pacientes analisados, sendo 169 (76,5 porcento) HIV-1, 43 (19,5 porcento) HIV-2 e 9 de (4,1 porcento) co-infectados pelo HIV-1 e pelo HIV-2, 67 porcento eram do sexo feminino. As medianas de idade foram de 34 (IQR = 1-75) e 47 (IQR = 12-84) para o HIV-1 e HIV-2, respectivamente. A infecção pelo HIV-1 é causada pelo subtipo G (36,6 porcento), CRF02_AG (30,6 porcento), subtipo F1, (9,7 porcento), URFs (10,4 porcento), subtipo B (5,2porcento), CRF05_DF (3,0 porcento), subtipo C (2,2 porcento), CRF06_cpx (0,7 porcento), CRF25_cpx (0,7 porcento) e CRF49_cpx (0,7 porcento), e todas as infecções por HIV-2 pertencem ao grupo A. De acordo com as análises filogeográficas e de origem do HIV, estimase que o HIV-2 foi o primeiro tipo viral introduzido em Cabo Verde e possui relações filogenéticas com sequências referências de Portugal...

HIV-1 and HIV-2 have been detected in Cape Verde since 1987, but little is known regarding the genetic diversity of these viruses in this archipelago, located near the West African coast. In this study, we characterized the molecular epidemiology of HIV-1 and HIV-2, analyzed the origin of the major clades of HIV introduced in CapeVerde and described the occurrence of drug resistance mutations (DRM) among antiretroviral therapy naïve (ARTn) patients and patients under treatment (ARTexp) from different Cape Verde islands. Blood samples, socio-demographic and clinicallaboratorydata were obtained from 221 HIV-positive individuals during 2010-2011.Genetic sequencing of the pol region (1300bp) was obtained and phylogenetic and bootscan analyses were performed for viral subtyping. The evolutionary and phylogeographic studies were performed using the program BEAST. HIV-1 and HIV-2 DRM were evaluated for ARTn and ARTexp patients using the Stanford HIVDatabase and HIV-GRADE e.V. Algorithm Homepage, respectively. Among the 221patients (169 [76.5 percent] HIV-1, 43 [19.5 percent] HIV-2 and 9 [4.1 percent] HIV-1/HIV-2 coinfections), 67 percent were female. The median ages were 34 (IQR=1-75) and 47(IQR=12-84) for HIV-1 and HIV-2, respectively. HIV-1 infections were due to subtypes G (36.6 percent), CRF02_AG (30.6 percent), F1 (9.7 percent), URFs (10.4 percent), B (5.2 percent), CRF05_DF (3.0 percent), C (2.2percent), CRF06_cpx (0.7 percent), CRF25_cpx (0.7 percent) and CRF49_cpx (0.7 percent), whereas all HIV-2 infections belonged to group A. According the HIV phylogeographic analyses, it is estimated that HIV-2 was the first viral type introduced in Cape Verde and has phylogenetic relationships with referral sequencesof Portugal...

Wikstroelide M potently inhibits HIV replication by targeting reverse transcriptase and integrase nuclear translocation / 中国天然药物

AIM@#To evaluate the anti-HIV activity and mechanism of action of wikstroelide M, a daphnane diterpene from Daphne acutiloba Rehder (Thymelaeaceae).@*METHODS@#The anti-HIV activities of wikstroelide M against different HIV strains were evaluated by cytopathic effect assay and p24 quantification assay with ELISA. The inhibitory effect of wikstroelide M on HIV reverse transcription was analyzed by real-time PCR and ELISA. The effect of wikstroelide M on HIV-1 integrase nuclear translocation was observed with a cell-based imaging assay. The effect of wikstroelide M on LEDGF/p75-IN interaction was assayed by molecular docking.@*RESULTS@#Wikstroelide M potently inhibited different HIV-1 strains, including HIV-1IIIB, HIV-1A17, and HIV-19495, induced a cytopathic effect, with EC50 values ranging from 3.81 to 15.65 ng·mL⁻¹. Wikstroelide M also had high inhibitory activities against HIV-2ROD and HIV-2CBL-20-induced cytopathic effects with EC50 values of 18.88 and 31.90 ng·mL⁻¹. The inhibitory activities of wikstroelide M on the three HIV-1 strains were further confirmed by p24 quantification assay, with EC50 values ranging from 15.16 to 35.57 ng·mL⁻¹. Wikstroelide M also potently inhibited HIV-1IIIB induced cytolysis in MT-4 cells, with an EC50 value of 9.60 ng·mL⁻¹. The mechanistic assay showed that wikstroelide M targeted HIV-1 reverse transcriptase and nuclear translocation of integrase through disrupting the interaction between integrase and LEDGF/p75.@*CONCLUSION@#Wikstroelide M may be a potent HIV-1 and HIV-2 inhibitor, the mechanisms of action may include inhibition of reverse trascriptase activity and inhibition of integrase nuclear translocation through disrupting the interaction between integrase and LEDGF/p75.

An HIV1/2 point of care test on sputum for screening TB/HIV co-infection in Central India - Will it work?

OBJECTIVE@#To determine whether the OraQuick® HIV-1/2 Assay (OraSure Technologies, Inc., Bethlehem, PA, USA) in sputum is a valid tool for HIV surveillance among TB patients.@*METHODS@#A cross sectional study was carried out on sputa of patients diagnosed with tuberculosis. Sputa were tested for antibodies to HIV using OraQuick® HIV-1/2 Assay (OraSure Technologies, Inc., Bethlehem, PA, USA). The results were compared with results of serum ELISA.@*RESULTS@#Compared to serum ELISA, the OraQuick® HIV-1/2 Assay in sputum specimens reported 90% sensitivity (9/10) and 100% specificity (307/307), with a positive predictive value of 100% (95%CI: 66.37%-100.00%) and a negative predictive value of 99.68% (95%CI: 98.20%-99.99%).@*CONCLUSIONS@#This testing method may provide a useful strategy for conducting HIV surveillance in possible co-infected TB patients at peripheral centres. Since there is no investment on infrastructure, it may be possible for paramedical health professionals to carry out the test, particularly in areas with low HIV endemicity.

Evaluation of rapid tests for human immunodeficiency virus as a tool to detect recent seroconversion

The identification of recent HIV infection is important for epidemiological studies and to monitor the epidemic. The objective of this study was to evaluate two rapid tests that are easily available to the Brazilian scientific community for using as markers of recent HIV infection. The Rapid Test - HIV-1/2 Bio-Manguinhos (Bio-Manguinhos/Fiocruz, Brazil) and the Rapid Check HIV 1&2 (NDI-UFES, Center for Infectious Diseases, Universidade Federal do Espírito Santo) were tested, using 489 samples with HIV positive serology, from blood donors, previously classified as recent or long-term infection by serological testing algorithm for recent HIV seroconversion (STARHS) or LS-HIV Vitros assay methods. The samples were diluted prior to testing (1:50 and 1:100 for the Rapid Test - HIV-1/2 Bio-Manguinhos, and 1:500 and 1:600 for the Rapid Check HIV 1&2). Negative samples were considered recent infection, whereas those showing any color intensity were associated with long-term infection. The best dilutions were 1:100 for HIV-1/2 Bio-Manguinhos test (Kappa = 0.840; overall agreement = 0.93), and 1:500 for the Rapid Check HIV 1&2 (Kappa = 0.867; overall agreement = 0.94). The results suggest that both rapid tests can be used to detect recent seroconversion.

Human immunodeficiency virus type-2-A milder, kinder virus: An update.

Human immunodeficiency virus type-2 (HIV-2) belongs to the family retroviridae which is phylogenetically clusters with SIV SM from sooty mangabeys. This virus is morphologically similar to human immunodeficiency virus type-1 (HIV-1) but has got only a 40% homology at the nucleotide level. There is a distinct geographical distribution of HIV-2 unlike HIV-1. There are currently eight subtypes/groups identified with subtype/group A responsible for the majority of infections. HIV-2 shows a considerable difference in the course of the disease. Clinical, haematological and immunological evaluation of individuals infected with HIV-2 has shown the virus to be less pathogenic than HIV-1 although the exact mechanism underlying this difference is not well defined. Similar to HIV-1, the HIV-2 isolates also showed distinct replicative and cytopathic characteristics. The transmission rate for HIV-2 compared to HIV-1 is very low both by heterosexual route and mother to child transmission. The clinical signs and symptoms of immunodeficiency associated with HIV-2 are similar to the ones seen among the HIV-1-infected individuals and they can also progress to AIDS. It is naturally resistant to NNRTI and hence the diagnosis become important as it affects the treatment strategy. Similar to HIV-1, HIV-2 strains of infected individuals also show mutations that can cause drug resistance. The current evidence suggests that there is no protective effective for HIV-2 against HIV-1.

Prevalence of HIV among blood donors in a tertiary care centre of north India.

Background & objectives: India has the second highest HIV population in the world with about 2.5-3.0 million cases. HIV-2 cases among general and blood donor population have also been reported mostly from west and south India. This single centre study was carried out to observe the HIV-1 and HIV-2 prevalence among blood donors from north India. Methods: A total of 2,04,677 people were screened for the presence of HIV infection over the 11 year period (1999 to 2009). Till 2004, a third generation ELISA kit was used. From 2005 till January 2009 all tests were done using the fourth generation ELISA kit which detected the presence of HIV-1 P24 antigen and anti-HIV antibodies. From February 2009 onwards, the kits used were Genscreen ULTRA HIV Ag- Ab Assay. Results: A total of 506 (0.247%) donors were found to be repeat reactive for HIV. Of these, 486 (96%) donors tested using the Western blot were found positive for HIV-1 infection. Twenty (4%) donors showed a negative Western blot result, none of the donors were found reactive for HIV-2 infection. Interpretation & conclusions: The prevalence of HIV was 0.249 per cent among blood donors of north India. No HIV-2 case was found among the studied blood donor population indicating that it is not a threat currently.

Genotypical diversity of HIV clades and central nervous system impairment / Diversidade genotípica dos subtipos de HIV e comprometimento do sistema nervoso central

The central nervous system (CNS) and the immune system are considered major target organs for HIV infection. The neurological manifestations directly related to HIV are acute viral meningitis, chronic meningitis, HIV associated dementia, vacuolar myelopathy and involvement of the peripheral nervous system. Changes in diagnosis and clinical management have changed the aspect of HIV infection so that it is no longer a fatal disease, and has become a chronic disease requiring sustained medical management. After HAART the incidence of most opportunistic infections, including those affecting the CNS, has dropped markedly. Some studies suggest that neurological involvement of infected patient occur with different frequency, depending on HIV subtype involved in the infection. Subtype C may have reduced neuroinvasive capacity, possibly due to its different primary conformation of HIV transactivating regulatory protein (Tat), involved in monocyte chemotaxis. This review focus on physiopathologic aspects of HIV infection in CNS and its correlation with HIV clades.

O sistema nervoso central (SNC) e o sistema imunológico são considerados os principais órgãos alvo na infecção pelo HIV. As manifestações neurológicas diretamente relacionadas ao HIV são meningites virais aguda e crônica, demência associada ao HIV, mielopatia vacuolar e envolvimento do sistema nervoso periférico. Mudanças no diagnóstico e sobrevida têm mudado o aspecto da infecção pelo HIV, não mais considerada uma doença fatal e sim crônica. Após HAART, a incidência da maioria das doenças oportunistas, incluindo aquelas que afetam o SNC, reduziu-se significativamente. Alguns estudos sugerem que o envolvimento de pacientes infectados ocorre com frequência diferente, dependendo do subtipo de HIV. O subtipo C apresenta uma capacidade reduzida de neuroinvasão, possivelmente devido a conformação primária da sua proteína reguladora da transativação (Tat), que perde sua capacidade quimiotáxica. Esta revisão aborda aspectos fisiopatológicos da infecção do HIV no SNC e subtipos de HIV.

Infección diseminada por Penicillium marneffei en un paciente HIV-positivo: Primera observación en la República Argentina / Disseminated infection due to Penicillium marneffei related to HIV infection: First observation in Argentina

Se presenta el primer caso humano de peniciliosis por Penicillium marneffei observado en la República Argentina. El paciente era un joven de 16 años, HIV-positivo, procedente de un área rural del sur de China. El paciente fue internado en el Hospital "F. J. Muñiz" por padecer una neumonía grave con insufciencia respiratoria aguda. El agente causal fue aislado de un lavado broncoalveolar y se lo observó en un citodiagnóstico de piel. La identifcación de P. marneffei fue confrmada por las características fenotípicas del aislamiento y la amplifcación del ADNr. El enfermo padecía una infección muy avanzada por HIV que condujo a la aparición simultánea de infecciones por citomegalovirus, Pneumocystis jirovecii y procesos bacterianos nosocomiales. Este complejo cuadro derivó en una evolución fatal.

The frst case observed in Argentina of AIDS-related human penicillosis is herein presented. The patient was a six- teen year-old young man coming from a rural area of southern China. He was admitted at the F. J. Muñiz Hospital of Buenos Aires city with severe pneumonia and adult respiratory distress. Penicillium marneffei was isolated from bronchoalveolar lavage fuid and was microscopically observed in a skin cytodiagnosis. P. marneffei identifcation was confrmed by rRNA amplifcation and its phenotypic characteristics. The patient suffered an advanced HIV infection and also presented several AIDS-related diseases due to CMV, nosocomial bacterial infections and Pneumocystis jirovecii which led to a fatal outcome.

Characteristics, immunological response & treatment outcomes of HIV-2 compared with HIV-1 & dual infections (HIV 1/ 2) in Mumbai.

Background & objectives: Information available on HIV-2 and dual infection (HIV-1/2) is limited. This study was carried out among HIV positive individuals in an urban referral clinic in Khar, Mumbai, India, to report on relative proportions of HIV-1, HIV-2 and HIV-1/2 and baseline characteristics, response to and outcomes on antiretroviral treatment (ART). Methods: Retrospective analysis of programme data (May 2006-May 2009) at Khar HIV/AIDS clinic at Mumbai, India was done. Three test algorithm was used to diagnose HIV-1 and -2 infection. Standard ART was given to infected individuals. Information was collected on standardized forms. Results: A total of 524 individuals (male=51%; median age=37 yr) were included in the analysis over a 3 year period (2006-2009) - 489 (93%) with HIV-1, 28 (6%) with HIV-2 and 7(1%) with dual HIV-1/2 infection. HIV-2 individuals were significantly older than HIV-1 individuals (P<0.001). A significantly higher proportion of HIV-2 patients and those with dual infections had CD4 counts <200 cells/µl compared to HIV-1. HIV-2 individuals were more likely to present in WHO Clinical Stage 4. Of the 443 patients who were started on ART, 358 (81%) were still alive and on ART, 38 (8.5%) died and 3 were transferred out. CD4 count recovery at 6 and 12 months was satisfactory for HIV-1 and HIV-2 patients on protease inhibitor based regimens while this was significantly lower in HIV-2 individuals receiving 3 nucleoside reverse transcriptase inhibitors. Interpretation & conclusions: In an urban HIV clinic in Mumbai, India, HIV-2 and dual infections are not uncommon. Adaptation of the current national diagnostic and management protocols to include discriminatory testing for HIV types and providing access to appropriate and effective ART regimens will prevent the development of viral resistance and preserve future therapeutic options.

Sero-prevalence of viral Co-infections in HIV infected children of northern India.

Objective. To assess the prevalence of viral co-infections in HIV infected children. Methods. Children born to HIV seropositive parents and those children who were suspected to be HIV infected based on clinical presentation by the pediatrician were screened for HIV –1 and 2 antibodies as per National Aids Control Organization (NACO) guidelines. Those found to be seropositive for HIV infection were further tested for Hepatitis B&C, Herpes simplex virus and Human cytomegalovirus infection. Results. Among 803 children screened, 101 were found positive for HIVantibodies. Among the five viral markers tested, HCMV IgG was positive in 88 children (87.1%). HCMV IgM was positive in 35 cases (34.6%). HBsAg tested positive in 30 children, while anti-HCV IgM was reactive in 27 cases. IgM anti- HSV antibodies were observed positive in 59 (58.4%) cases. Both hepatitis virus coinfection (HBsAg and anti- HCV IgM antibodies) was observed in 10 HIV positive children, while both Herpesviridae family viruses (HCMV -IgM antibodies and HSV -IgM antibodies) were positive in 30 cases (29.7%). Conclusion. Viral co-infections are significantly higher in HIV positive children, which adds to significant mortality and morbidity and should therefore be screened in all HIV positive children for timely treatment in order to improve the quality of life and better survival of HIV infected children.

Assessment of a self-deferral form for screening blood donors, Chiang Mai University Hospital, Thailand.

A self-deferral form has been used to screen Chiang Mai University Hospital blood donors and was improved in 2005. It has never been evaluated. The study aimed to assess the self-deferral form procedures in detecting infected donors. Sera from 5,083 donors, who passed the self-deferral screening form, were tested with the routine immuno-assays (serology) for HIV 1 and 2 antibodies, P24 antigen, HCV antibodies, HBV surface antigen, and syphilis. Antibody negative sera were also tested individually with the the Procleix Ultrio Assay for HIV-1 DNA, HCV RNA, and HBV DNA. The donors who had discrepant results between serology and NAT were evaluated with additional tests, including a more sensitive Alternative Nucleic Acid Test, AntiBcore IgM, AntiBcore IgG, HBsAg and Anti HBs. Among 5,083 donors, 331 (6.5%) had at least one positive marker. In multiple logistic regression analysis, the statistically significant factors (adjusted odds ratio and 95% CI) for infection were age 30 years or below [1.45 (1.03, 2.03)], male gender [2.73 (1.64, 4.56)], primary school or lower education [1.56 (1.09, 2.23)], first-time donation [1.82 (1.25, 2.67)], and frequent donation [0.80 (0.70, 0.92)]. The safest donors were females, older than 30 years, with an education more than primary school, and frequent donation. Because of missing responses to some sensitive questions, there remains a need for further improvement of the self-deferral form.

Evaluation of the human immunodeficiency virus type 1 and 2 antibodies detection in dried whole blood spots (dbs) samples / Pesquisa de anticorpos contra o vírus da imunodeficiência humana tipos 1 e 2 em amostras de sangue seco coletadas em papel filtro

Human Immunodeficiency Vírus Type 1 and 2 antibodies detection was performed in 457 dried whole blood spots samples (S&S 903). Q-Preven HIV 1+2 was the screening test used. The results were compared with the gold standard serum tests by ELISA (Cobas Core e Axsym HIV1/2 gO) and imunofluorescence was the definitive confirmatory test. The samples were obtained from the Hospital Nossa Senhora da Conceição in Porto Alegre, RS - Brazil, through whole blood transfer to filter paper card and sent to Caxias do Sul, RS - Brazil where the tests were performed. The dried whole blood spot stability was evaluated with two different panels. The first one was composed of five negative and five positive samples stored at room temperature, 4 ºC, -20 ºC and -70 ºC, while the second was composed of two negative and three positive samples stored at 37 ºC (humidity <50 percent). Each sample was screened every week for six weeks. These measurement results didn't show variation during the study period. The detected sensibility was 100 percent, specificity was 99.6 percent, the positive predictive value was 99.5 percent and negative predictive values were 100 percent. The results demonstrated high performance characteristics, opening a new perspective of dried whole blood spot utilization in HIV screening diagnosis.

Foram realizados 457 testes para detectar anticorpos contra o Vírus da Imunodeficiência Humana tipos 1 e 2, em amostras de sangue total seco coletadas em papel filtro (S&S 903), com o teste de triagem Q-Preven HIV 1+2, comparando-se com os resultados dos testes de triagem no soro (Cobas Core e Axsym HIV1/2 gO), sendo a imunofluorescência indireta o teste confirmatório. As amostras foram obtidas no Hospital Conceição em Porto Alegre, pela transferência de sangue total para cartão de papel filtro e encaminhadas para Caxias do Sul para a realização dos testes. Foi analisada a estabilidade da amostra em papel filtro com a utilização de dois painéis: o primeiro com cinco amostras negativas e cinco positivas armazenadas por seis semanas à temperatura ambiente, 4 ºC, -20 ºC e -70 ºC; o segundo com duas negativas e três positivas armazenadas por seis semanas com avaliações semanais a 37 ºC (umidade <50 por cento). Os resultados de todas as amostras testadas foram mantidos. A sensibilidade foi de 100 por cento, a especificidade de 99,6 por cento, o valor preditivo positivo de 99,5 por cento e o valor preditivo negativo de 100 por cento. O excelente desempenho observado na análise da utilização de sangue seco em papel filtro, abre uma nova perspectiva no diagnóstico da infecção pelo HIV.

Virus herpes humano 8: detección en células mononucleares de sangre periférica de pacientes con la infección por el virus de inmunodeficiencia humana: con/sin sarcoma de kaposi / Human herpes virus 8: detection in peripheral blood mononuclear cells from patients with human immunodeficiency virus infection with/without kaposi sarcoma

El ácido nucleico proveniente del virus herpes humano 8 esta presente en las células mononucleares de sangre periférica de un 50% a 90% de los pacientes con sarcoma de Kaposi, y 7% a 10% de los pacientes con la infección por el virus de inmunodeficiencia humana sin sarcoma de Kaposi. Nosotros estudiamos la prevalencia del virus herpes humano 8 en células mononucleares de sangre periférica provenientes de pacientes con la infección por el virus inmunodeficiencia humana con/sin sarcoma de Kaposi. Setenta y seis pacientes con la infección por el virus de inmunodeficiencia humana sin sarcoma de Kaposi y 15 pacientes con sarcoma de Kaposi asociado a la infección por el virus de inmunodeficiencia humana se incluyeron en el estudio. El ácido desoxirribonucleico se extrajo utilizando el método de fenol/cloroformo. El ácido desoxirribonucleico fue amplificado a través de la reacción en cadena de la polimerasa utilizando las sondas KS1 y KS2 específicas para el ORF26 del virus herpes humano 8. Las reacciones se consideraron positivas sólo si los productos de la región esperada de 233 pares de bases. Ninguno de los pacientes con la infección por el virus de inmunodeficiencia humana mostró la presencia de virus herpes humano 8 en las células mononucleares de sangre periférica, y después de un seguimiento de 2 años, ninguno ha desarrollado sarcoma de Kaposi. El virus herpes humano 8 se detectó en las células mononucleares de sangre periférica del 20% de los pacientes con sarcoma de Kaposi. Todos los pacientes pertenecían al grupo de "altó riesgo", y eran varones homosexuales. Ninguno recibió transfusiones sanguíneas. Estos datos preliminares sugieren que la prevalencia del virus herpes humano 8 en las células mononucleares de sangre periférica de los pacientes con la infección por el virus de inmunodeficiencia humana con/sin sarcoma de Kaposi es probablemente baja en comparación con pacientes provenientes de Estados Unidos y Europa

The nucleic acid of human herpes virus 8 is present in the peripheral blood mononuclear cells of between 50% and 90% of Kaposi sarcoma patients, 7% and 10% of human immunodeficiency virus infected patients without Kaposi sarcoma. We studied the prevalence of human herpes virus 8 in peripheral blood mononuclear cells from patients with human immunodeficiency virus infection with/without Koposi sarcoma. Seventy-six patients with human immunodeficiency virus infection without Kaposi sarcoma associated with human immunodeficiency virus infection were included. Desoxibonucleico acid was extracted from the sample by a standard phenol/chloroform extraction procedure. Desoxirribonucleico acid was polimerase chain reaction amplified using Kaposi sarcoma 1 and Kaposi sarcoma 2 primers specific for the human herpes virus 8 ORF26. Polimerase chain reaction were considered positive only if the polimerase chain reaction products hybridized in the expected 233 by region. None of human immunodeficiency virus infected patients showed the presence of human herpes virus 8 in the peripheral blood mononuclear cells, and after a follow-up of 2 years, none has developed Kaposi sarcoma. Human herpes virus 8 was detected in the peripheral blood mononuclear cells from 20% of patients with Kaposi sarcoma. All patients belonged to a "high risk group", were male and homosexuals. None received blood transfusion. These preliminary data suggest that the prevalence of human herpes virus 8 in peripheral blood mononuclear cells from human immunodeficiency virus infected patients with/withoul Kaposi sarcoma is probably low in comparison with patients from EE.UU and Europe

GTVseq: A Web-based Genotyping Tool for Viral Sequences

Genotyping Tool for Viral SEQuences (GTVseq) provides scientists with the genotype information on the viral genome sequences including HIV-1, HIV-2, HBV, HCV, HTLV-1, HTLV-2, poliovirus, enterovirus, flavivirus, Hantavirus, and rotavirus. GTVseq produces alternative and additive genotype information for the query viral sequences based on two different, but related, scoring methods. The genotype information produced is reported in a graphical manner for the reference genotype matches and each graphical output is linked to the detailed sequence alignments between the query and the matched reference sequences. GTVseq also reports the potential 'repeats' and/or 'recombination' sequence region in a separated window. GTVseq does not replace completely other well-known genotyping tools such as NCBI's virus sequence genotyping tool (http://www.ncbi. nlm.nih.gov/projects/genotyping/formpage.cgi), but provides additional information useful in the confirmation or for further investigation of the genotype(s) for the newly isolated viral sequences.