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1.
J Med Virol ; 93(5): 3069-3076, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-33554363

RESUMEN

The implementation of rapid diagnostic tests (RDTs) may enhance the efficiency of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) testing, as RDTs are widely accessible and easy to use. The aim of this study was to evaluate the performance of a diagnosis strategy based on a combination of antigen and immunoglobulin M (IgM) or immunoglobulin G (IgG) serological RDTs. Plasma and nasopharyngeal samples were collected between 14 March and 11 April 2020 at hospital admission from 45 patients with reverse transcription polymerase chain reaction (RT-PCR) confirmed COVID-19 and 20 negative controls. SARS-CoV-2 antigen (Ag) was assessed in nasopharyngeal swabs using the Coris Respi-Strip. For IgM/IgG detection, SureScreen Diagnostics and Szybio Biotech RDTs were used in addition to laboratory assays (Abbott Alinity i SARS-CoV-2 IgG and Theradiag COVID-19 IgM enzyme-linked immunosorbent assay). Using the Ag RDT, 13 out of 45 (29.0%) specimens tested positive, the sensitivity was 87.0% for cycle threshold (Ct ) values ≤25% and 0% for Ct values greater than 25. IgG detection was associated with high Ct values and the amount of time after the onset of symptoms. The profile of isolated IgM on RDTs was more frequently observed during the first and second week after the onset of symptoms. The combination of Ag and IgM/IgG RDTs enabled the detection of up to 84.0% of COVID-19 confirmed cases at hospital admission. Antigen and antibody-based RDTs showed suboptimal performances when used alone. However when used in combination, they are able to identify most COVID-19 patients admitted in an emergency department.


Asunto(s)
Anticuerpos Antivirales/sangre , Antígenos Virales/sangre , Prueba Serológica para COVID-19/métodos , COVID-19/diagnóstico , SARS-CoV-2/aislamiento & purificación , Adulto , Anciano , Anciano de 80 o más Años , COVID-19/virología , Ensayo de Inmunoadsorción Enzimática , Femenino , Hospitalización , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Masculino , Persona de Mediana Edad , Nasofaringe/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Pruebas Serológicas/métodos
2.
J Viral Hepat ; 27(5): 540-543, 2020 05.
Artículo en Inglés | MEDLINE | ID: mdl-31895489

RESUMEN

Diagnostic of acute hepatitis A virus (HAV) infection is based on the detection of anti-HAV IgM without testing for the pathogen itself. We evaluated the usefulness of HAV RNA testing for confirmation of acute hepatitis A and to provide indications about the level of HAV replication in HIV-positive and HIV-negative subjects during an unprecedented outbreak of HAV observed in France in 2017. HAV RNA was detected in 38 out of 41 (92.6%) subjects with a clinical diagnosis of acute hepatitis A, whereas nine cases tested positive for anti-HAV IgM in whom the diagnosis of acute hepatitis A was not retained were found negative for HAV RNA. All subjects in the control group were also tested negative for HAV RNA. HAV viremia was correlated to ALT peak (r = .64; P < .0001). HIV-infected patients have similar HAV RNA levels but were less likely to have prolonged international normalized ratio of prothrombin time when compared to the HIV-uninfected group (P = .016), suggesting a less severe course of acute hepatitis. HAV RNA was detected in the serum of most of the patients with acute hepatitis A, indicating that the direct detection of HAV can be used to confirm hepatitis A in patients tested positive for anti-HAV IgM antibodies. Nucleic acid tests should serve more broadly during the diagnosis workup of acute hepatitis A to improve the predictive values of HAV in vitro diagnostic tests and to confirm acute hepatitis A in patients tested positive with IgM with moderate or low S/CO values.


Asunto(s)
Hepatitis A/diagnóstico , ARN Viral/sangre , Pruebas Diagnósticas de Rutina , Brotes de Enfermedades , Francia , Hepatitis A/epidemiología , Anticuerpos de Hepatitis A/sangre , Humanos , Inmunoglobulina M/sangre
3.
BMC Infect Dis ; 20(1): 49, 2020 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-31941464

RESUMEN

BACKGROUND: Hepatitis B is a major concern in Africa, especially in HIV-infected patients. Unfortunately, access to hepatitis B virus (HBV) testing and adequate treatment remains a challenge in the continent. We investigated HBV testing, treatment, and virologic suppression in HIV-infected patients followed up as part of Cameroon's national antiretroviral programme. METHODS: A cross-sectional survey was performed in adult patients receiving antiretroviral therapy (ART) in 19 hospitals in the Centre and Littoral regions in Cameroon. The proportions of patients tested for hepatitis B surface antigen (HBsAg) prior to the study were compared among all study hospitals using the Chi-square test. The association of individual and hospital-related characteristics with HBV testing and virologic suppression was assessed using multilevel logistic regression models. RESULTS: Of 1706 patients (women 74%, median age 42 years, median time on ART 3.9 years), 302 (17.7%) had been tested for HBsAg prior to the study. The proportion of HBV-tested patients ranged from 0.8 to 72.5% according to the individual hospital (p < 0.001). HBV testing was lower in women (adjusted odds ratio [aOR] 0.64, 95% confidence interval [CI] 0.46-0.89, p = 0.010) and higher in patients who initiated ART in 2010 or later (aOR 1.66, 95% CI 1.23-2.27, p < 0.001). Of 159 HBsAg-positive patients at the time of the study (9.3%), only 97 (61.0%) received Tenofovir + Lamivudine (or Emtricitabine). Of 157 coinfected patients, 114 (72.6%) had a HBV viral load < 10 IU/mL. HBV suppression was higher in patients with a HIV viral load < 300 copies/mL (aOR 3.46, 95% CI 1.48-8.09, p = 0.004) and lower in patients with increased ALT level (aOR 0.86 per 10 IU/mL increase, 95% CI 0.75-0.97, p = 0.019). CONCLUSIONS: A substantial proportion of HIV/HBV coinfected patients were at higher risk of liver disease progression. Improving the management of HBV infection in the routine healthcare setting in Africa is urgently required in order to achieve the 2030 elimination targets. Micro-elimination of HBV infection in people living with HIV could be an easier and cost-effective component than more widely scaling up HBV policies.


Asunto(s)
Coinfección/tratamiento farmacológico , Infecciones por VIH/tratamiento farmacológico , Virus de la Hepatitis B/genética , Hepatitis B/tratamiento farmacológico , Hepatitis B/virología , Respuesta Virológica Sostenida , Adulto , Antirretrovirales/uso terapéutico , Camerún , Estudios Transversales , Femenino , Estudios de Seguimiento , Genotipo , Antígenos de Superficie de la Hepatitis B/sangre , Humanos , Masculino , Persona de Mediana Edad , Salud Pública , Carga Viral
4.
Bioanalysis ; 9(5): 427-434, 2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-28220711

RESUMEN

AIM: Hemoglobin A1c (HbA1c) is a widely recognized analyte for diagnosing and monitoring diabetes. Dried blood spot (DBS) constitutes a useful alternative to blood collection by venipuncture. Analytical and clinical validation of DBS use is, however, necessary before implementation. Results/methodology: HbA1c levels from whole blood or DBS from a cohort patients with diabetes were compared. DBS specimens were stable at ambient temperature. HbA1c detection on DBS was accurate, robust, and the correlation and agreement with whole blood values was excellent. CONCLUSION: This study provides for the first time a complete method comparison and validation under the ISO15189 guideline using an automated HPLC system. This approach constitutes, therefore, a useful tool for diagnosing diabetes.


Asunto(s)
Cromatografía Líquida de Alta Presión , Cromatografía por Intercambio Iónico , Pruebas con Sangre Seca , Hemoglobina Glucada/análisis , Adulto , Anciano , Automatización , Biomarcadores/sangre , Recolección de Muestras de Sangre , Estudios de Casos y Controles , Diabetes Mellitus Tipo 1/sangre , Diabetes Mellitus Tipo 1/patología , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/patología , Femenino , Hemoglobina Glucada/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Estabilidad Proteica
5.
Mediterr J Hematol Infect Dis ; 8(1): e2016003, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26740864

RESUMEN

BACKGROUND: Treatment of hepatitis C virus (HCV) infection based on peginterferon-α (pegIFNα) and ribavirin induces important changes in cytokine release and T cell activation. OBJECTIVE: Immune response to pegIFNα-ribavirin therapy was explored in patients coinfected by HCV and HIV. METHODS: Concentrations of 25 cytokines and CD8(+) T cell activation were monitored in HCV/HIV coinfected patients classified as sustained virological responders (SVR, n=19) and non-responders (NR, n=11). RESULTS: High pretreatment concentrations of IP-10 (CXCL-10) and MCP-1 (CCL-2) were associated with a poor anti-HCV response. PegIFNα-ribavirin therapy increased CD8(+) T cell activation and induced significant changes in levels of eleven cytokines related to both Th1 and Th2 responses in SVR (IL-1ß, IL-1RA, IL-4, IL-5, IL-6, IL-7, IL-12p40/70, IL-13, IP-10, eotaxin, MCP-1) but of only six cytokines in NR (IL-1ß, IL-2, IL-5, IL-12p40/70, IL-13, eotaxin). The highest rise in MIP-1ß and MCP-1 levels was observed four weeks after anti-HCV treatment initiation in SVR compared to NR (p=0.002 and p=0.03, respectively), whereas a decrease in IL-8 concentration was associated with treatment failure (p= 0.052). CONCLUSIONS: Higher and broader cytokine responses to pegIFNα-ribavirin therapy were observed in SVR patients compared to NR. Changes in IL-8, MIP-1ß, and MCP-1 serum concentrations may be associated with efficacy of pegIFNα- and ribavirin-based therapies in patients coinfected by HCV and HIV.

6.
Antimicrob Agents Chemother ; 60(3): 1608-14, 2015 Dec 28.
Artículo en Inglés | MEDLINE | ID: mdl-26711757

RESUMEN

Hepatitis E virus (HEV) can lead to chronic infection in solid-organ transplant patients. Ribavirin is efficient for treatment of chronically infected patients. Recently, the1634R mutation in the HEV polymerase has been associated with treatment failure. However, it is unclear if this mutation can be used as a prognostic marker of treatment outcome. We studied the prevalence of the 1634R mutation in the HEV polymerase of patients starting ribavirin therapy, the influence of the 1634R variants on the viral response, the frequency of the 1634R mutation in patients whose treatment failed, and its impact on ribavirin retreatment. We analyzed pretreatment samples from 63 solid-organ transplant patients with chronic hepatitis E using deep sequencing; 42 patients had a sustained virologic response (SVR), and 21 were non-SVR patients. We detected the 1634R variant by deep sequencing in 36.5% (23/63) of the patients (proportions, 1.3 to 100%). The 1634R variant was detected in 31.0% (13/42) of baseline plasma samples from patients with SVR and in 47.6% (10/21) in the other patients (P = 0.2). The presence of this mutation did not influence the initial decrease in viral RNA. Lastly, a second prolonged ribavirin treatment led to SVR in 70% of the patients who initially did not have SVR, despite the presence of the 1634R variant. We conclude that the presence of the 1634R variant at ribavirin initiation does not lead to absolute ribavirin resistance. Although its proportion increased in patients whose treatment failed, the presence of the 1634R variant did not compromise the response to a second ribavirin treatment.


Asunto(s)
Antivirales/uso terapéutico , Virus de la Hepatitis E/efectos de los fármacos , Virus de la Hepatitis E/genética , Hepatitis E/tratamiento farmacológico , ARN Polimerasa Dependiente del ARN/genética , Ribavirina/uso terapéutico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Secuencia de Bases , Niño , Femenino , Marcadores Genéticos , Hepatitis E/virología , Humanos , Masculino , Persona de Mediana Edad , Mutación/genética , ARN Viral/genética , Análisis de Secuencia de ARN , Resultado del Tratamiento , Adulto Joven
7.
J Virol Methods ; 201: 24-30, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24560781

RESUMEN

In-house developed real-time PCR (qPCR) techniques could be useful conjunctives to the management of hepatitis B virus (HBV) infection in resource-limited settings with high prevalence. Two qPCR assays (qPCR1 and qPCR2), based on primers/probes targeting conserved regions of the X and S genes of HBV respectively, were evaluated using clinical samples of varying HBV genotypes, and compared to the commercial Roche Cobas AmpliPrep/Cobas TaqMan HBV Test v2.0. The lower detection limit (LDL) was established at 104 IU/ml for qPCR1, and 91 IU/ml for qPCR2. Good agreement and correlation were obtained between the Roche assay and both qPCR assays (r = 0.834 for qPCR1; and r = 0.870 for qPCR2). Differences in HBV DNA load of > 0.5 Log10 IU/ml between the Roche and the qPCR assays were found in 49/122 samples of qPCR1, and 35/122 samples of qPCR2. qPCR1 tended to underestimate HBV DNA quantity in samples with a low viral load and overestimate HBV DNA concentration in samples with a high viral load when compared to the Roche test. Both molecular tools that were developed, used on an open real-time PCR system, were reliable for HBV DNA detection and quantitation. The qPCR2 performed better than the qPCR1 and had the additional advantage of various HBV genotype detection and quantitation. This low cost quantitative HBV DNA PCR assay may be an alternative solution when implementing national programmes to diagnose, monitor and treat HBV infection in low- to middle-income countries where testing for HBV DNA is not available in governmental health programmes.


Asunto(s)
ADN Viral/análisis , Virus de la Hepatitis B/aislamiento & purificación , Hepatitis B/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Carga Viral/métodos , Cartilla de ADN/genética , ADN Viral/genética , Antígenos de Superficie de la Hepatitis B/genética , Virus de la Hepatitis B/genética , Humanos , Sondas de Oligonucleótidos/genética , Transactivadores/genética , Proteínas Reguladoras y Accesorias Virales
8.
Clin Chem Lab Med ; 52(5): 649-55, 2014 May.
Artículo en Inglés | MEDLINE | ID: mdl-24150208

RESUMEN

BACKGROUND: The use of dried blood spots on filter paper is well documented as an affordable and practical alternative to classical venous sampling for various clinical needs. This technique has indeed many advantages in terms of collection, biological safety, storage, and shipment. Amyloid ß (Aß) peptides are useful cerebrospinal fluid (CSF) biomarkers for Alzheimer disease diagnosis. However, Aß determination is hindered by preanalytical difficulties in terms of sample collection and stability in tubes. METHODS: We compared the quantification of Aß peptides (1-40, 1-42, and 1-38) by simplex and multiplex ELISA, following either a standard operator method (liquid direct quantification) or after spotting CSF onto dried matrix paper card. RESULTS: The use of dried matrix spot (DMS) overcame preanalytical problems and allowed the determination of Aß concentrations that were highly commutable (Bland-Altman) with those obtained using CSF in classical tubes. Moreover, we found a positive and significant correlation (r2=0.83, Pearson coefficient p=0.0329) between the two approaches. CONCLUSIONS: This new DMS method for CSF represents an interesting alternative that increases the quality and efficiency in preanalytics. This should enable the better exploitation of Aß analytes for Alzheimer's diagnosis.


Asunto(s)
Péptidos beta-Amiloides/líquido cefalorraquídeo , Ensayo de Inmunoadsorción Enzimática , Fragmentos de Péptidos/líquido cefalorraquídeo , Enfermedad de Alzheimer/diagnóstico , Biomarcadores/líquido cefalorraquídeo , Pruebas con Sangre Seca , Humanos
9.
Gut ; 63(9): 1490-500, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24153249

RESUMEN

OBJECTIVE: Adult primary human hepatocytes (PHHs) support the complete infection cycle of natural HCV from patients' sera. The molecular details underlying sera infectivity towards these cells remain largely unknown. Therefore, we sought to gain a deeper comprehension of these features in the most physiologically relevant culture system. DESIGN: Using kinetic experiments, we defined the optimal conditions to infect PHH and explored the link between cell organisation and permissivity. Based on their infectivity, about 120 sera were classified in three groups. Concentration of 52 analytes was measured in 79 selected sera using multiplexed immunobead-based analyte profiling. RESULTS: PHH permissivity towards HCV infection negatively correlated with cell polarisation and formation of functional bile canaliculi. PHH supported HCV replication for at least 2 weeks with de novo virus production. Depending on their reactivity, sera could be classified in three groups of high, intermediate or low infectivity toward PHH. Infectivity could not be predicted based on the donors' clinical characteristics, viral load or genotype. Interestingly, highly infectious sera displayed a specific cytokine profile with low levels of most of the 52 tested analytes. Among them, 24 cytokines/growth factors could impact hepatocyte biology and infection efficiency. CONCLUSIONS: We identified critical factors leading to efficient PHH infection by HCV sera in vitro. Overall, we showed that this cellular model provides a useful tool for studying the mechanism of HCV infection in its natural host cell, selecting highly infectious isolates, and determining the potency of drugs towards various HCV strains.


Asunto(s)
Hepacivirus/patogenicidad , Hepatocitos/virología , Adulto , Biomarcadores/metabolismo , Técnicas de Cultivo de Célula/métodos , Línea Celular , Células Cultivadas , Citocinas/metabolismo , Hepacivirus/metabolismo , Hepatocitos/fisiología , Humanos , Cinética , Modelos Inmunológicos , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Suero/virología
10.
Clin Chem Lab Med ; 51(10): 1897-909, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23740687

RESUMEN

The analysis of blood spotted and dried on a matrix (i.e., "dried blood spot" or DBS) has been used since the 1960s in clinical chemistry; mostly for neonatal screening. Since then, many clinical analytes, including nucleic acids, small molecules and lipids, have been successfully measured using DBS. Although this pre-analytical approach represents an interesting alternative to classical venous blood sampling, its routine use is limited. Here, we review the application of DBS technology in clinical chemistry, and evaluate its future role supported by new analytical methods such as mass spectrometry.


Asunto(s)
Recolección de Muestras de Sangre/normas , Enfermedades Transmisibles/diagnóstico , Pruebas con Sangre Seca/estadística & datos numéricos , Enfermedades Metabólicas/diagnóstico , Preservación Biológica/normas , Adulto , Cromatografía Liquida , Pruebas Enzimáticas Clínicas , Pruebas con Sangre Seca/instrumentación , Pruebas con Sangre Seca/métodos , Pruebas Genéticas/métodos , Humanos , Recién Nacido , Espectrometría de Masas , Reacción en Cadena de la Polimerasa
11.
PLoS One ; 7(3): e32143, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22403628

RESUMEN

BACKGROUND: The decline in hepatitis B virus surface antigen (HBsAg) may be an early predictor of the viral efficacy of Hepatitis B virus (HBV) therapy. The HBsAg levels obtained by different immunoassays now need comparing and the relationships between levels of HBsAg and HBV DNA alongside HBsAg and genotype must be evaluated. METHODOLOGY/PRINCIPAL FINDINGS: HBsAg levels were compared among 80 patients using the Abbott Architect assay, a commercial immunoassay approved for HBsAg detection and quantitation, and three other assays derived from immunoassays approved for HBsAg detection (manufactured by Diasorin, Bio-Rad and Roche). Good correlation was found between the Abbot vs. Diasorin, Bio-Rad and Roche assays with narrow 95% limits of agreement and small mean differences: -0.06 to 0.11, -0.09 log(10) IU/mL; -0.57 to 0.64, -0.04 log(10) IU/mL; -0.09 to 0.45, -0.27 log(10) IU/mL, respectively. These agreements were not affected by genotypes A or D. HBsAg was weakly correlated with HBV DNA, whatever the HBsAg assay used: Abbott, ρ = 0.36 p = 0.001, Diasorin ρ = 0.34, p = 0.002; Bio-Rad ρ = 0.37, p<0.001; or Roche ρ = 0.41, p<0.001. This relationship between levels of HBsAg and HBV DNA seemed to depend on genotypes. Whereas HBsAg (Abbott assay) tended to correlate with HBV DNA for genotype A (ρ = 0.44, p = 0.02), no such correlation was significant for genotypes D (ρ = 0.29, p = 0.15). CONCLUSION/SIGNIFICANCE: The quantitation of HBsAg in routine clinical samples is comparable between the reference assay and the adapted assays with acceptable accuracy limits, low levels of variability and minimum discrepancy. While HBsAg quantitation is not affected by HBV genotype, the observed association between levels of HBsAg and HBV DNA seems genotype dependent.


Asunto(s)
Genotipo , Antígenos de Superficie de la Hepatitis B/sangre , Virus de la Hepatitis B/genética , Virus de la Hepatitis B/fisiología , Inmunoensayo/métodos , Replicación Viral , Adulto , ADN Viral/sangre , ADN Viral/genética , Antígenos e de la Hepatitis B/sangre , Humanos
12.
Antivir Ther ; 17(2): 321-6, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22290198

RESUMEN

BACKGROUND: In Africa, most HIV-HBV-coinfected patients on antiretroviral therapy (ART) receive an anti-HBV lamivudine monotherapy that has been shown in northern countries to lead to frequent emergence of drug resistance. We assessed the HBV prevalence and the rate and pattern of lamivudine-resistant HBV mutations in Cameroonian HIV-infected, ART-treated patients. METHODS: A cross-sectional survey was performed in 2006-2007 at the HIV/AIDS outpatient clinic of the Central Hospital in Yaoundé, Cameroon. Plasma samples were tested as appropriate for hepatitis B surface antigens, antibodies to hepatitis B core, HBV DNA, genotypes and lamivudine-resistant polymerase mutations. RESULTS: Of 552 adult patients (71% women, median age 38 years), 290 had received lamivudine-based ART for 12 months and 262 for 24 months. No patient had received tenofovir. The prevalence of hepatitis B surface antigen was 9.8%. Overall, 26% of seropositive patients had an HBV DNA level >40 IU/ml. Genotypes A and E were identified. Polymerase resistance mutations were detected in 14% and 60% of patients at months 12 and 24, respectively. CONCLUSIONS: This study supports both WHO recommendations of screening for HBV before initiation of ART and of using ART containing tenofovir and either lamivudine or emtricitabine in HIV-HBV-coinfected patients in Africa.


Asunto(s)
Farmacorresistencia Viral/genética , Infecciones por VIH/tratamiento farmacológico , Virus de la Hepatitis B/efectos de los fármacos , Hepatitis B/tratamiento farmacológico , Lamivudine/farmacología , Adulto , Fármacos Anti-VIH/farmacología , Fármacos Anti-VIH/uso terapéutico , Secuencia de Bases , Camerún , Coinfección/tratamiento farmacológico , Coinfección/virología , Estudios Transversales , ADN Viral/sangre , Femenino , Productos del Gen pol/genética , Infecciones por VIH/complicaciones , VIH-1/efectos de los fármacos , VIH-1/genética , Hepatitis B/complicaciones , Antígenos de Superficie de la Hepatitis B/sangre , Virus de la Hepatitis B/genética , Humanos , Lamivudine/uso terapéutico , Masculino , Persona de Mediana Edad , Análisis de Secuencia de ADN
13.
BMC Public Health ; 10: 105, 2010 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-20193053

RESUMEN

BACKGROUND: Coinfection with hepatitis B virus (HBV) or hepatitis C virus (HCV) in HIV-infected patients receiving a commonly used nevirapine-based antiretroviral therapy is a major concern for African clinicians owing to its high prevalence, the infrequent testing and treatment of viral hepatitis, and the impact of liver disease on the tolerability and effectiveness of anti-HIV treatment. We compared the hepatotoxicity and the immunological, virological and clinical effectiveness of a nevirapine-based antiretroviral therapy between patients infected with HIV only and patients coinfected with hepatitis B or C virus in Cameroon. METHODS: A retrospective cohort study was conducted among HIV-1-infected patients. Plasma HBV DNA and HCV RNA were tested in positive or indeterminate samples for HBsAg or HCV antibodies, respectively. All patients received nevirapine and lamivudine plus stavudine or zidovudine. RESULTS: Of 169 HIV-1-infected patients with a median baseline CD4 count of 135 cells/mm3 (interquartile range [IQR] 67-218), 21% were coinfected with HBV or HCV. In coinfected patients, the median viral load was 2.47 x 107 IU/mL for HBV (IQR 3680-1.59 x 108) and 928 000 IU/mL for HCV (IQR 178 400-2.06 x 106). Multivariate analyses showed that the risk of hepatotoxicity was 2-fold higher in coinfected patients (p < 0.01). The response to antiretroviral therapy was however comparable between monoinfected and coinfected patients in terms of CD4 cell count increase (p = 0.8), HIV-1 viral load below 400 copies/mL (p = 0.9), death (p = 0.3) and death or new AIDS-defining event (p = 0.1). Nevirapine was replaced by a protease inhibitor in 4 patients owing to hepatotoxicity. CONCLUSION: This study suggests that the nevirapine-based antiretroviral therapy could be used safely as first-line treatment in patients with low CD4 cell count in Africa despite frequent coinfections with HBV or HCV and infrequent testing of these infections. Although testing for HBV and HCV should be systematically performed before initiating antiretroviral therapy, transaminases elevations at baseline or during treatment should be a decisive argument for testing when hepatitis status is unknown.


Asunto(s)
Fármacos Anti-VIH/uso terapéutico , Infecciones por VIH/tratamiento farmacológico , Hepatitis B/complicaciones , Hepatitis C/complicaciones , Hígado/efectos de los fármacos , Nevirapina/uso terapéutico , Adulto , Fármacos Anti-VIH/efectos adversos , Recuento de Linfocito CD4 , Camerún , Enfermedad Hepática Inducida por Sustancias y Drogas/prevención & control , Estudios de Cohortes , Femenino , Infecciones por VIH/complicaciones , Humanos , Pruebas de Función Hepática , Masculino , Nevirapina/efectos adversos , Estudios Retrospectivos , Factores de Riesgo , Carga Viral
14.
Hepatology ; 51(3): 752-8, 2010 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-20043287

RESUMEN

UNLABELLED: We investigated the performance of dried blood spots (DBS) in hepatitis C virus (HCV) diagnosis using modified commercial tests. Paired DBS and serum samples were collected from 200 patients: 100 patients with anti-HCV antibodies (anti-HCV), including 62 patients with detectable serum HCV RNA, and 100 patients without anti-HCV. The DBS sample consisted of three drops of approximately 50 microL of whole blood applied to a paper card, which was then stored at -20 degrees C within 48 hours of collection. Using the Ortho HCV 3.0 enzyme-linked immunosorbent assay kit on DBS, we observed both a specificity and sensitivity of 99% in detecting anti-HCV. HCV RNA was detected on DBS in 60/62 (97%) patients with detectable serum HCV RNA, which was then successfully quantified in 55 samples (89%) using the Cobas TaqMan HCV test. A good correlation was observed between the DBS HCV RNA concentration and the serum level (r(2) = 0.95; P < 0.001). HCV genotyping was successfully performed on DBS samples, with a full concordance between the 14 paired DBS and serum samples (genotypes 1-4). CONCLUSION: This study presents DBS as a reliable alternative to serum specimens for detecting anti-HCV, quantifying HCV RNA and genotyping HCV. DBS may increase the opportunities for HCV testing and treatment follow-up in hard-to-reach individuals.


Asunto(s)
Anticuerpos contra la Hepatitis C/sangre , Hepatitis C/sangre , Hepatitis C/diagnóstico , Estudios de Factibilidad , Pruebas Hematológicas/métodos , Hepacivirus/genética , Hepacivirus/inmunología , Humanos , Técnicas de Diagnóstico Molecular , ARN Viral/análisis , Sensibilidad y Especificidad , Pruebas Serológicas
15.
J Clin Microbiol ; 45(9): 3077-81, 2007 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-17581929

RESUMEN

The VERSANT HCV RNA 3.0 (bDNA), COBAS AmpliPrep/COBAS TaqMan HCV, and Abbott ART HCV RealTime assays were compared for hepatitis C virus RNA quantification in 158 clinical specimens (genotypes 1 to 5). RNA values differed significantly between methods (P < 0.0001), and mean titer differences ranged from 0.01 to 0.50 log(10) IU/ml depending on the genotypes.


Asunto(s)
Hepacivirus/genética , Hepatitis C/virología , Técnicas de Diagnóstico Molecular/métodos , ARN Viral/sangre , Suero/virología , Carga Viral/métodos , Humanos , Juego de Reactivos para Diagnóstico
16.
Acta Derm Venereol ; 87(1): 14-7, 2007.
Artículo en Inglés | MEDLINE | ID: mdl-17225009

RESUMEN

Human herpes virus 8 infection is the primary and necessary factor in the development of Kaposi's sarcoma, but is not sufficient per se to trigger the onset of the disease. In order to search for virological cofactors associated with the occurrence of the disease, we investigated the prevalence of active infection by two newly discovered viruses, hepatitis G virus and TT virus, among patients with classical Kaposi's sarcoma. Serum of 24 patients with Mediterranean Kaposi's sarcoma was investigated using polymerase chain reaction and compared with that of 68 healthy subjects. Cutaneous samples from patients with Kaposi's sarcoma and healthy subjects were investigated for TT virus DNA. No patient had serum markers for hepatitis G virus. TT virus DNA was present in the serum of 21/24 (87.5%) patients and 32/68 (47%) controls (p=0.002). TT virus DNA was present in the lesional skin of 5/18 patients with Kaposi's sarcoma (27.7%), but not in the skin of controls. TT virus might play a role as a cofactor in the clinical emergence of Kaposi's sarcoma in patients infected with Human herpes virus 8, perhaps by immunosuppressive effects or by a common transmission pathway for these two viruses.


Asunto(s)
Infecciones por Virus ADN/epidemiología , Sarcoma de Kaposi/virología , Torque teno virus/aislamiento & purificación , Adulto , Anciano , Anciano de 80 o más Años , Infecciones por Virus ADN/virología , ADN Viral/análisis , Femenino , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Prevalencia , Sarcoma de Kaposi/epidemiología
17.
J Immunol Methods ; 315(1-2): 144-52, 2006 Aug 31.
Artículo en Inglés | MEDLINE | ID: mdl-16959261

RESUMEN

To improve the investigation of the role of human memory B lymphocytes following hepatitis B virus (HBV) infection or vaccination, we developed a method to characterize circulating memory B cells specific to hepatitis B surface antigen (HBsAg). Our approach combined: (1) purification of CD19+ cells, (2) CD40-CD40L polyclonal stimulation, and (3) enumeration of memory B cells differentiated into anti-HBs antibody (Ab)-secreting cells (HBs-SCs) by a HBs-ELISPOT assay. In this way, HBs-SCs were detected in 17 HBsAg-vaccinated and nine HBV-immunized subjects including four individuals with serum anti-HBs Ab levels < 10 mIU/ml, but not in six controls. IgG+, IgA+ plus IgM+ HBs-SCs, representing 5-1736 cells/10(6) circulating B cells and 0.02-0.58% of total immunoglobulin-SCs generated by the B cell polyclonal stimulation, were counted by an Ig two-colour ELISPOT assay. In addition, anti-HBs Abs were found in 8/15 supernatants recovered from B cell cultures which contained HBs-SCs, suggesting that the HBs-ELISPOT assay is more reliable in tracking HBsAg-specific memory B cells than ELISA measurement of anti-HBs Abs secreted in supernatants. This new approach could be useful to explore the presence and the longevity of HBsAg-specific memory B cells in vaccinated and immunized subjects, in chronic HBV infection and after liver transplantation for HBV-related disease.


Asunto(s)
Linfocitos B/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Antígenos de Superficie de la Hepatitis B/inmunología , Vacunas contra Hepatitis B/inmunología , Hepatitis B/inmunología , Memoria Inmunológica , Células Productoras de Anticuerpos/metabolismo , Antígenos CD40/metabolismo , Estudios de Casos y Controles , Diferenciación Celular , Células Cultivadas , Antígenos de Superficie de la Hepatitis B/sangre , Humanos , Inmunoglobulinas/sangre
18.
J Acquir Immune Defic Syndr ; 38(3): 305-9, 2005 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-15735449

RESUMEN

CCR5 antagonists represent promising anti-HIV agents. Yet, if the CCR5 chemokine receptor plays a positive role in hepatitis C virus (HCV) infection, CCR5 antagonists might be contraindicated in HCV/HIV-coinfected subjects. Therefore, we tested the hypothesis that the level of T-cell surface CCR5 expression, which might determine the intensity of HCV-specific T-cell recruitment into the liver, and thereby the efficiency of the anti-HCV response, could determine HCV disease evolution. For this purpose, we compared CCR5 density, measured by quantitative flow cytometry at the surface of nonactivated (human leukocyte antigen-D-related [HLA-DR]-) T cells of 51 HCV/HIV patients, with HCV load, serum aminotransferase levels, and liver histology (inflammatory activity, fibrosis, and rate of fibrosis progression). DR-CD4+ T-cell surface CCR5 density, which correlated with DR-CD8+ T-cell surface CCR5 density and was stable over time in HCV/HIV-coinfected individuals, did not correlate with any of the biologic parameters of HCV infection analyzed and was not linked to the capacity to clear the virus. In conclusion, we failed to demonstrate any impact of interindividual variability in T-cell surface CCR5 density on HCV infection, which would have argued against the use of CCR5 antagonists in HIV/HCV-coinfected subjects.


Asunto(s)
Infecciones por VIH/complicaciones , Hepatitis C/complicaciones , Hepatitis C/inmunología , Receptores CCR5/análisis , Linfocitos T/inmunología , Adulto , Antagonistas de los Receptores CCR5 , Antígenos CD4/análisis , Linfocitos T CD4-Positivos/inmunología , Antígenos CD8/análisis , Linfocitos T CD8-positivos/inmunología , Femenino , Citometría de Flujo , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/inmunología , Antígenos HLA-DR/análisis , Hepacivirus/aislamiento & purificación , Hepacivirus/fisiología , Hepatitis C/patología , Hepatitis C/virología , Humanos , Hígado/patología , Cirrosis Hepática/patología , Masculino , Persona de Mediana Edad , Transaminasas/sangre , Carga Viral
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